QUIZ 1A LEC Question 6

Question 1 A device for rapid pipetting made of spring activated

plunger in a precision bore with disposable tip
How close a measurement to the true value is called
o Volumetric pipet
o Precision
o Micropipette
o Significant
o Serological pipette
o Accuracy
o Dropper
o Estimate

Question 2
Question 7
A substance of known composition, value of which is
established by an analytical procedure Precision pipet considered most accurate is (di ko sure
potek. Kasi sabi diba pinakaccurate yung volumetric eh
o Reagent
to deliver ang volumetric. Tapos basta hindi Ostwald
o Blank
sagot sa isa)
o Control
o Standard o Ostwald-Folin pipet
o Serological pipet
o TC volumetric pipet
Question 3
o TD volumetric pipet
The chemical reagent that has the highest purity for
laboratory use
Question 8
o USP
In the proper use of a volumetric pipet calibrated to
o Analytical grade
deliver (TD) one should
o Reagent grade
o Technical grade o Blow out the solution immediately
o Drain the content but do not blow out
o Wipe the outside wall of the pipet
Question 4 o Rinse the content several times to mix the
solution
Glassware of choice for dilution and preparation of
solutions that are made to definite volume

o Volumetric flask Question 9

o Graduated cylinder
Plastic wares are useful in the laboratory because of
o Erlenmeyer flask
what characteristic(s)?
o Automatic pipet
o All options are correct
o Shatter proof
Question 5
o High impact and tensile strength
A one gram equivalent weight of the solute in one liter o Chemically inert
of solution is expressed as

o One normal solution

o One percent solution
o One molar solution
o One milliequivalent per liter
Question 10 Question 15

Solutions that are used in keeping the pH relatively It has no markings down to the tip allows the content of
constant the pipet to be blown out

o Standards o Serological pipet

o Buffers o To deliver pipet
o Enzymes o Mohr pipet
o Water o Transfer pipet

Question 11 Question 16

One gram equivalent weight of an element equals the Grades of chemicals include all of the following except
gram molecular weight divided by the
o Commercial grade
o Valence o Industrial grade
o Volume o Analytic reagent
o Dilution o Chemically pure
o Mole o

Question 17

Question 12 Water from which minerals have been removed is called

A molar solution contains o Charcoal activated water

o One gram equivalent weight of solute per liter
of solution
o None of the options are correct
o One gram molecular weight of solute per liter of
solution
o One gram of solute to 99 gram of solvent
Question 13
What happens to a decimal place in the number when
you are converting from a smaller unit to a larger unit?
o Reagent grade water
o Distilled water
o Deionized water
Question 18
When should the sides of the pipet be wiped off?
o After reaching the lower meniscus of the mark
o Only when using TC or TD pipet
o When using serological pipet and pipette
o Before lowering the meniscus to the calibrated
mark

o Converting doesn’t require placing

o The decimal place moves to the left
Question 19
o The decimal place moves to the right
o The decimal place doesn't move at all Of a volumetric pipette mark with a ring near the mouth
end, the solution must be
Question 14
o Rinsed out
20°℃ is __ °F
o Blown out
o 68 o Drain out
o 53 o Left in the pipet before rinsing out with distilled
o 86 water
o 25
Question 20 Question 25

Reagent grade Type III water is used in All of the following describes positive displacement
micropipet except
o Highly sensitive procedures
o Rinsing or washing of glassware o With disposable piston
o Preparation of culture media o For volatile sample
o Preparation of buffer o Piston is a permanent part of the pipet
o No air cushion

Question 21

When using a micropipettor, if you push to the second

stop to fill it you will get

o Either too much or too little solution

o The correct amount of solution
o Too much solution
o Too little solution

Question 22

Which of these is a correct colligative property?

o Boiling point depression

o Density
o Vapor pressure lowering
o Freezing point elevation

Question 23

The boiling point of a solution is that of a pure solvent

o Higher than
o Equal to
o Lower than
o Less than

Question 24

Borosilicate glass is commercially known as

o Corning
o Pyrex
o Soft glass
o Soda lime glass
QUIZ 2A LEC Question 5

Question 1 Which condition is a common cause of stray light?

In absorption spectrophotometry A. Dispersion from second-order spectra

A. Percent transmittance is directly proportional to B. Improper wavelength calibration

concentration
C. Misaligned source lamp
B. Absorbance is directly proportional to concentration
D. Unstable source lamp voltage
C. Percent transmittance is directly proportional to the
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dehydrates and atomizes a sample using

A. A thermoelectric semiconductor
Question 6
B. An electron gun
Which type of filter is best for measuring stray light?
C. A graphite capillary furnace
A. Sharp cutoff
D. A thermospray platform
B. Wratten

C. Neutral density
Question 3
D. Didymium
Which type of monochromator produces the purest
monochromatic light in the UV ra
Question 7
A. A prism and a variable exit slit
Which component is required in a spectrophotometer
B. A sharp cutoff filter and a variable exit slit
in order to produce a spectral absorbance curve?
C. Interference filters and a variable exit slit
A. Photodiode array
D. A diffraction grating and a fixed exit slit
B. Multiple monochromators
https://quizlet.com/202476638/clinical-chemistry-
C. Laser light source
instruments-mls-review-flash-cards/
D. A reference optical beam
Question 4
https://www.flashcardmachine.com/spectrophotometr
Which photodetector is most sensitive to low levels of
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C. Photodiode

D. Photomultiplier tube
Question 8 Question 12

Optical density is referred to as Optimally, where should your unknown concentrations

fall in your standard curve?
A. Absorbance
A. To the left of your lowest data point.
B. Transmittance
B. To the right of your highest data point.
C. Emission
C. Close the midpoint between your known
D. Percent
concentrations.

D. When possible, above the line of best fit.

Question 9
https://quizizz.com/admin/quiz/5e7e8cf1d05b8b001b5
According to the Beer-Lambert law, absorbance is f79a8/spectrophotometer

A. Inversely proportional to the concentration Question 13

B. Directly proportional to the log of the concentration A spectrophotometer can be used to

C. Directly proportional to the transmittance A. Determine the presence of a substance in a solution.

D. Directly proportional to the concentration of the B. Measure the concentration of an unknown solution.
solution
C. Identify an unknown substance

D. Determine the molecular composition of an unknown

Question 10 substance.

If the following reaction is known to obey Beer's law,

the X axis is the concentration of the solution and the Y
Question 14
axis is
Which of the following lamps provides a continuous
A. None of the options are correct
spectrum of radiant energy in the visible and near UV
B. Wavelength regions?

C. Peak weight A. Tungsten-filament

D. Percent transmittance B. Mercury vapor

C. Hydrogen

Question 11 D. Deuterium

Following the standard Beer's Law, given diluted volume

of 5, 10, 20 and 40 standard, what will be the
Question 15
appearance of the graph? (not sure huhu)
Which of the following isolates light within a narrow
A. A straight line on semilog paper
region of the spectrum?
B. A straight line on percent transmission
A. Monochromator
C. A curve line on semilog paper
B. Photovoltaic cell
D. A straight line on a linear paper
C. Photomultiplier tube

D. Detector
Question 16 D. micrometers (µm)

The reagent blank corrects for absorbance caused by

A. The color of reagents Question 20

B. Sample turbidity When performing spectrophotometer quality assurance

checks, what is the helmium oxide glass filter used to
C. All of these options are correct
assess?
D. Bilirubin and hemolysis
A. Wavelength accuracy
https://freezingblue.com/flashcards/327301/preview/c
B. Stray light
hem01-instrumentation-and-methods
C. Linearity

D. Absorbance accuracy
Question 17

A standard calibration curve is used in the laboratory for

Question 21
A. Calibration of reagents
Which of the following formulas is an expression of the
B. Maintenance of a satisfactory performance
Beer-Lambert law that is routinely applied to
C. Standardization of reagents spectrophotometric analysis?

D. Determination of the concentration of an unknown A. Asx Cs/Cu = Au

B. A 2-log%T

Question 18 C. Cux Cs/As = Au

Which of the following is not descriptive of a D. Aux Cs/As = Cu

photomultiplier tube?
Question 22

Which of the following may be associated with

A. Cannot be read with a chopper reflectance spectrophotometry as it relates to the dry
reagent slide technique?
B. Amplifies the initial signal received

C. Must be shielded from stray light

A. Reflectance values are linearly proportional to
D. Emits electrons proportionally to initial light transmission values
absorbed
B. Light projected to the slide at 180-degree angle
https://quizlet.com/155817851/ch-1-success-clinical-
chemistry-437-q-flash-cards/ C. Dye concentration directly proportional to
reflectance
Question 19
D. Unabsorbed, reflected light detected by
What units of measurement are traditionally applied to photodetector
radiant energy in the visible portion of the
electromagnetic spectrum? https://www.brainscape.com/flashcards/multiple-
choice-question-rt-3007910/packs/4841322?page=1
A. nanometers (nm)
https://quizlet.com/405594324/success-in-clinical-
B. millimeters (mm) laboratory-science-clinical-chemistry-instrumentation-
C. centimeters (cm) and-analytical-principles-1-35-flash-cards/
Question 23

Matching Type

Checked with standard absorbing solution or filters -

Linearity

Any wavelength outside of the band transmitted - Stray

light

Demonstrated when a change in concentration results –

Wavelength
QUIZ 1A LAB Question 7

Question 1 Pipets are used to measure and dispense small amounts

of liquids. You should draw the liquid into the pipet
If an acid is splashed on your skin, wash at once with
using your mouth.
A. plenty of water
False
B. soap
Question 8
C. weak base
Work areas should be kept clean and tidy.
D. oil
True
Question 2
Question 9
Ten percent sodium hydroxide is used for
All chemicals in the lab are to be considered dangerous.
A. removing grease
True
B. cleaning new pipettes
Question 10
C. routine washing
All work surfaces should be sanitized at the end of the
D. removing blood clots shift with a solution

Question 3 o 10% bleach

o Concentrated
You are allowed to enter the chemical o 5% bleach
preparation/storage area any time you need to get an o 5% phenol
item.
Question 11
False
Decontaminate hands after
Question 4
o contact with patient's skin
All unauthorized experiments are prohibited. o contact with blood or body fluids
A. True o all statements are correct
o removing gloves
B. False
Question 12
Question 5
When you finish working with chemicals, biological
Chipped or cracked glassware is okay to use specimens, and other lab substances, always
A. True o treat your hands with skin lotion
B. False o wipe your hands on a towel
o wipe your hands on your clothes
Question 6 o wash your hands thoroughly with soap and
water
Laboratory work can be started immediately upon
entering the laboratory even if the instructor is not yet Question 13
present.
If a piece of equipment is not working properly, stop,
A. True turn it off, and tell
B. False o the custodian
o your best friend in the class
o your lab partner
 o the instructor Question 19

Question 14 If a lab experiment is not completed, you should

Why are safety symbols important? o make up some results

o come in during lunch and finish while eating
o They tell you how to do an experiment
lunch
o They tell you specific dangers
o discuss the issue with your instructor
o So you know what to do
o sneak in after school and work alone
o They are cool
Question 20
Question 15
After completing an experiment, all chemical wastes
In a laboratory, the following should not be worn.
should be
o loose clothing
o taken home
o dangling jewelry
o disposed of according to your instructor's
o all items are correct
directions
o sandals
o dumped in the sink
Question 16 o left at your lab station for the next class

Long hair in the laboratory must be Question 21

o cut short If you do not understand a direction or part of a lab

o held away from the experiment with one hand
o always neatly groomed
o tied back or kept entirely out of the way with a
hair band, hairpins, or other confining device
Question 17
procedure, you should
o figure it out as you do the lab
o ask the instructor before proceeding
o skip it and go on to the next part
o try several methods until something works

The term Standard Precautions refers to Question 22

o Treating only blood or blood-tinged specimens Approved eye protection devices (such as goggles) are
as infectious worn in the laboratory
o Assuming that every direct contact with a body
o any time chemicals, heat or glassware are used
fluid is not infectious
o To avoid eye strain
o All statements are correct
o to improve your vision
o Treating all specimens as if they are infectious
o only if you don't have corrective glasses
Question 18
Question 23
When gathering glassware and equipment for an
If a laboratory fire erupts, immediately
experiment, you should
o run for the fire extinguisher
o clean any glassware that appears dirty
o open the windows
o read all directions carefully to know what
o notify your instructor
equipment is necessary
o throw water on the fire
o examine all glassware to check for chips or
cracks Question 24
o All statements are correct
Flammable materials, like alcohol, should never be
dispensed or used near
 o another student QUIZ 2A LAB
o an open flame
Question 1
o an open door
o a sink In the proper use of a volumetric pipet calibrated to
deliver (TD), one should
Question 25
A. Drain the content but do not blow out
All infectious materials should be properly disposed in a
___ plastic bag and submitted to the lab technician for B. Blow out the solution immediately
proper disposal.
C. Wipe the outside of the pipet with cotton
o blue
o yellow D. Rinse the content several times to mix the solution
o green Question 2
o red
A piper with double ring at the mouth piece indicates
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D. To be rinsed several times with the diluent and then

blow out

http://mt-lectures.blogspot.com/2011/11/introduction-
to-clinical-chemistry.html

Question 3

The following are quality control tips in pipetting except

A. Air must not be aspirated during pipeting

B. Use pipets that are clean, no chips and correct size

for the volume to be measure

C. Drain the pipet in a slightly slanting position

D. The tip of the pipet must be wiped with gauze or a

tissue after coming from the solution

Question 4

Device for rapid pipetting made of spring activated

plunger in a precision bore with disposable tip

o Serological pipet
o Dropper
o Micropipette
o Pasteur pipette
Question 5 Question 9

A pipet should be wiped off In spectrophotometry, which of the following is a

mathematical expression of the relationship between
o After lowering the meniscus to the calibration
absorbance and transmittance?
mark
o Never if it is a volumetric pipet o Au/Cu = As/Cs
o Before lowering the meniscus to the calibration o A = log %T
mark o QA-2-log %T
o Only if it is a TC (to contain) pipet o A=abc

https://www.studystack.com/flashcard-1429021 Question 10

Question 6 A solution that has a transmittance of 1.0% would have

an absorbance of
When using an automatic pipet calibrated TC (to
contain) o 99%
o 1.0
o It must be made horizontally when filling
o 2.0
o Depress the piston to the first stop and the
o 1%
second stop to empty
o Depress the piston to the second stop to fill and https://quizlet.com/255591129/harr-mls-review-
the first stop to empty chemistry-51-instrumentation-flash-cards/ ditto na
o Depress the piston to the second top to both fill lahat hayup
and empty

Question 7
Question 11
When using a TC pipet in getting blood sample, how will
A spectrophotometer is set at zero optical density by
you dispense the blood sample?
using a
o Slowly drain the sample and rinse the pipet
o Control
o This is used as a serological pipet
o Serum sample
o Blow out all the blood sample
o Blank
o Slowly drain the blood sample
o Standard
Question 8

To properly use a volumetric pipet calibrated "to

Question 12
deliver" (TD), one should (not sure)
Parts of a spectrophotometer are
o Rinse out the contents several times
o Drain the contents but do not blow out o Tungsten lamp, cuvet, prism, phototube
o Drain the contents to the lowest etched mark in o Tungsten lamp, slit, filter, cuvet,
the volumetric pipet phototube.galvanometer
o Wipe the outside following the delivery of the o Tungsten lamp, slit, grating or prism, slit, cuvet,
contents phototube, galvanometer
o Tungsten lamp, grating or prism, cuvet,
phototube, galvanometer
Question 13 Question 17

According to Beer's law, the absorbance is Which of the following components determines the
wavelength of light that will pass through the sample
o Inversely proportional to the concentration
cuvette in a spectrophotometer?
o Directly proportional to the concentration
o Inversely proportional to the square of the o Detector
concentration o Light source
o Proportional to the square of the concentration o Monochromator
o Potentiometer

Question 14
Question 18
Spectrophotometer isolate a narrow band pass by
means of Which type of photodetector has a linear array that
allows it to respond to a specific wavelength resulting in
o Filter
complete UV/visible spectrum analysis?
o Prisms and layer cells
o Prisms and filter o Photodiode array
o Prisms and grating o Phototube
o Photovoltaic cell
Question 15
o Photomultiplier tube
Which of the following lists represents the light path
Question 19
through the components of a spectrophotometer
beginning immediately after the light source? Which of the following pipets are classified as transfer
pipets?
o Monochromator, entrance slit, sample cuvet,
exit slit, detector o Micropipets
o Entrance slit, monochromator, exit slit, sample o Ostwald-Folin
cuvet, detector o Mohr
o Monochromator, sample cuvet, entrance slit, o Serological
detector, exit slit
Question 21
o Entrance slit, sample, cuvet. exit slit.
monochromator, detector Which of the following pipets is calibrated to the tip?
Question 16 o None of these pipets
o Serological
Which of the following blanks is used to compensate for
o Mohr
absorption of the color of the test sample before
o Both pipets
reagents are added?
Question 22
o Water bank
o Alcohol blank Matching Type
o Reagent blank
o Sample blank Pipette with a long cylindrical tube drawn out to a tip
and is calibrated for blow out and in uniform fractional
volume of measurement

Volumetric Pipette
Pipette calibrated for the total volume of liquid and
must be washed out completely or must be rinsed
thoroughly

To continue pipette

Pipette calibrated for blow out of any small amount of

liquid remaining in the tip after delivery

To deliver pipette

Pipette calibrated for one specified volume and used for

pipetting

Serological pipette
MT–CC 1.1 (Clinical Chemistry) LEC Basic
Principles and Practices
Clinical Chemistry
 Basic science that utilizes the specialty of
chemistry to study human beings in various
stages of health and disease
 An applied science when analyses are
performed on body fluids or tissue specimens
to provide important information for the
diagnosis and treatment of disease (provide
us kung healthy ba yung tao or meron siyang
disease)
 The test must be performed accurately if the
results of lab analyses are to be useful to the
physician in diagnosing and treating patients
o Accuracy – it represents how close your
measurements come to its true value.
So dapat malapit na malapit yung
results mo sa true value.
o Precision – how close a series of
measurements of the same thing are to
each other. Not necessarily na naging
very close unlike accuracy.
Measurements that are imprecise do not
properly identify random errors, and
that can yield a widespread result
aiding now patient diagnosis and
treatment.
 The use of high quality analytical methods
and instrumentation is essential to lab work
 Concerns:
 Units of measure
 Basic laboratory supplies
 Introductory laboratory mathematics
 Specimen collection, processing and
reporting
Units of Measure
 Laboratory result consists of two components:
1. Number related to the actual value
2. Label identifying the units
3. Example: In glucose determination, the result
obtained is 120mg/dL, 120 is the actual value
and mg/dL is the label/unit
4. The unit defines the physical quantity of
dimension such as mass, length, time or volume
 Systeme International d’unites (SI) – adopted
internationally in 1960, is preferred in clinical
laboratories and is the only system employed in
many countries.
1. Devised to provide to provide the global
scientific community with a uniform method of
describing physical quantities. (to avoid confusion)
2. SI system units (SI) are based on the metric system
3. Several subclassifications exist within the SI
system, one of which is the basic unit
4. Basic units of measurement
 Metre (meter) - length
 Second - time
 Ampere – electric current
 Candela – luminous intensity
 Kilogram - mass
 Mole – quantity of a substance
 Kelvin – dynamic temperature
5. Used most often
 Length
 Mass
 Volume – quantity of a substance in mole
 A derived unit, as the name suggests, is a derivative or a
mathematical function describing one of the basic units
1. Example: meters per second (m/s), used to
express velocity
Some Non SI Units that have become acceptable
 Long-standing units – hour, minute, day, gram, liter
and plane angles expressed as degrees
1. These units, although widely used, can’t
technically be categorized as either basic or
derived SI units.
 Standard prefixes when added to a given basic unit,
can indicate decimal fractions or multiple of that
unit
1. Example: 0.001 L can be expressed using the
prefix milli, or 10-3. And since it requires
moving the decimal point three places to the
right, it can be written as 1 millimeter, or
abbreviated as 1mL.
2. It may also be written in scientific notation
as 1x10-3 L. Likewise, 1000 liters would use
the prefix of kilo (103) and could be written
as 1 kiloliter or expressed in scientific
notation as 1x103 L.
 When converting between prefixes, simply note the
relationship between them based on whether you
are changing to a smaller or larger prefix and the
incremental factor between them
1. Example:
- If converting from one liter (1.0x100 or
1) to milliliters (1.0x10-3 pr 0.001), the starting
unit is larger than the desired unit by a factor of
1000 or 103.
- This means that the decimal place would be
moved to the right of one (1) three places, so
1.0 liter (L) equals 1000 milliliters (mL).
- When changing 1000 milliliter (mL) to 1 liter (L),
the process is reversed and decimal point would
be moved three places to the left to become 1L.
 SI conversions

1. Convert to larger unit: move decimal point to
left
2. Convert to smaller unit: move decimal
point to the right
 The SI term for mass in kilogram, it is the only
basic unit that contains a prefix as part of its
naming convention.
 Generally, the standard prefixes for mass use the
term gram rather than kilogram.
MOST COMMON GRADES
 Reporting of lab results is often expressed in
terms of substance concentration (e.g. moles)
or the mass of a substance (e.g. mg/dL, g/dL,
g/L, mmol/L and IU) rather than SI units.
 It has been recommended that analytes be
reported using moles of solute per volume of
solution (substance concentration) and the
liter be used as the reference volume.
Reagents
 Reagent – any substance producing a chemical
reaction
 In a highly automated laboratory, there seems
to be little need for reagent preparation.
 Reagents are ready-to-use form or in a kit form
(i.e. all necessary reagents and respective
storage containers pre-package as a unit)
requiring only the addition of water or buffer
for reconstitution
 Everything is prepared when we use the reagent
kit.
 Periodically, especially in hospital laboratories
involved specialized analyses or method
validation, one may still face preparing
various reagents or solutions.
 As a result of deterioration of reagents, supply
and demand or the institution of cost-
containment programs --- prepare reagents in-
house
Therefore, a thorough knowledge of chemicals,
standards, solutions, buffers and water requirements is
necessary.
 Reagent quality control records must be retained for 5
years.
 Reagents shall be used and controlled according to the
manufacturer’s recommendations.
 All reagents and chemicals, including solvents and
materials used in tests and assays, should be of
appropriate quality.
 Reagent should be purchased from reputable, approved
suppliers and should be accompanied by the certificate of
analysis, and the material safety data sheet.
Chemicals
 Analytic chemicals exist in varying grades of purity:
1. Analytic reagent (AR)
2. Ultrapure
3. Chemically pure CP)
4. United States Pharmacopeia (USP)
5. National Formulary (NF) and
technical or commercial grade
You should know when to use this chemicals with a grades
of purity
 ACS grade
1. A chemical grade of highest purity and meets or
exceeds purity standards set by American Chemical
Society
2. Acceptable for food, drug or medicinal use and can
be used for ACS application or for general
procedures that require stringent quality
specification and a purity of ≥95%.
 Reagent grade
1. High purity generally equal to ACS grade and
suitable for use in many laboratories and analytical
applications
2. Acceptable for food, drug or medicinal use
 USP grade
1. A chemical grade of sufficient purity to meet or
exceed the requirements of USP
2. Acceptable for food, drug, or medicinal use; may be
used for most laboratory purposes
 NF grade
1. A grade of sufficient purity to meet or exceed
requirements of the National Formulary (NF)
2. The USP and the NF USP-NF jointly publish a book of
public pharmacopeial standard for chemical and
biological drug substances, dosage forms and
compounded preparations, medical devices and
dietary supplements
 Lab grade
1. A chemical grade of relatively high quality with
exact levels of impurities unknown
2. Usually pure enough for educational applications
3. While excellent for teaching and training, it is not
pure enough to be offered for food, drug or
medicinal use of any kind
 Purified grade
1. Also called pure or practical grade and
indicates good quality chemicals meeting
no official standard
2. Can be used in most cases for educational
applications
3. Not pure enough to be offered for food,
drugs or medicinal use of any kind
 Technical grade
1. Good quality chemical grade used for
commercial and industrial purposes
2. Not pure enough to be offered for food,
drug or medicinal use of any kind
2. TYPE 2: Purified water
 Workhouse of the water world, employed for
general laboratory use in things like culture
media preparation or buffer creation
3. TYPE 3: Primary grade water
 Water operating behind the scenes, used for non-
critical work like rinsing beakers, filling water
baths or feeding autoclaves
Solutions
 Mixtures in which soluble particles are completely
dissolved in a liquid or gas
 Made up of solute and solvent

Reference Materials
 Primary standard is a highly-purified chemical
that can be measured directly to produce a
substance of exact known concentration and
purity.
 Secondary standard is a substance of lower
purity, with its concentration determined by
comparison with a primary standard.

Water Specifications
 Reagent grade water (RGW)
1. Suitable for reagent and standard preparation
2. Most procedures use distilled water or
deionized water
 Distilled water
1. Purified to remove almost all organic materials
2. Water may be distilled more than once and
 Solution Properties
each distillation cycle will remove
1. Concentration
impurities
 Percent solution
 Deionized water
3 expressions of percent solution
1. Produced from distilled water using either
o Weight per weight (w/w)
an anion or cation exchange resin followed
o Volume per volume (v/v)
by replacement of the removed particles
o Weight per volume (w/v) – use g/dL instead
with hydroxyl or hydrogen ions, respectively of percent
 Reverse osmosis – pumps water across a semi- o For v/v solutions, it is recommended that
permeable membrane and produces RO water gram per decilitre (g/dl) be used instead of
 Water can also be purified by ultrafiltration, UV percentage or % (v/v)
light, sterilization or ozone treatment
 Lab requirements generally call for reagent
grade water that, according to the Clinical and
Laboratory Standards Institute (CLSI), is
classified into needed for its one of six categories
based on the specifications use rather the
method of purification or preparation
 Categories
1. Clinical laboratory reagent
2. Special reagent water
3. Instrument feed water
4. Water supplied by method of manufacturer
5. Autoclave and wash water  Normality
o Least likely encountered; used in chemical
6. Commercial bottled purified water
titrations & chemical reagent classification
 Types o Defined as the number of gram equivalent
1. TYPE 1 : Ultrapure water weights per 1L of solution.
 Used for highly sensitive procedures
like HPLC, AAS and mammalian cell
culture
 o Valence – number of units that can
combine with 1 mole of hydrogen ions for
acids and hydroxyl ions for bases and the
number of electrons exchanged in
oxidation-reduction reactions
o The number of atoms/elements that can
combine for a particular compound;
therefore the equivalent weight is the
gram combining weight of a material
o Normality is always equal to or greater
than the molarity of that compounds
o Normality was previously used for
reporting electrolyte values such as Na+,
K+, Cl- expressed as milliequivalent per
liter (mEq/L), however, has been
replaced with more familiar units of
millimoles/liter (mmol/L)
 Molarity
o Number of moles per 1L of solution
o One mole of a substance equal its gram
molecular weight (GMW), so the
customary units of molarity are
moles/liter
o The SI expression for concentration
should be represented as mol/L, mmol/L,
µmol/L, nmol/L
 Molality
-
Saturation
Dilute and unsaturated – relatively little

solute or one which has been made to
lower solute concentration per volume of
solvent as when making a dilution
 Concentrated solution– large quantity of
solute in solution
 Supersaturated – has an even greater
concentration of solute particles than a
saturated solution of the same substance
o Thermodynamically unstable - greater
concentration of solute particles
o Addition of a solute or mechanical agitation
disturbs the supersaturated solution,
resulting in crystallization of any excess
material out of solution
 Saturated – solution in which there is an
excess of undissolved solute particles
2. Colligative properties
o Properties of solutions that depend on the
ratio of the number of solute particles to the
number of solvent molecules in a solution,
and not on the nature of the chemical species
present
o The number ratio can be related to the
various units for concentration of a solution,
for example, molarity, molality, normality
(chemistry), etc.
o The assumption that solution properties are
independent of nature of solute particles of only
exact for ideal solutions and is approximate for
dilute real solutions.
o Colligative properties are a set of solution
properties that can be reasonably approximated
by assuming that the solution is ideal.
o Only properties which result from the dissolution
of nonvolatile solute in a volatile liquid solvent are
considered.
o They are essentially solvent properties which
are changed by the presence of the solute
o The solute particles displace some solvent
molecules in the liquid phase are therefore
reducing the concentration of the solvent, so
that colligative properties are independent of
the nature of the solute
o Affected by the
 Vapor pressure – pressure at which the
liquid solvent is in equilibrium with the
water vapor
o Vapor pressure lowering – the
 vapor pressure of a solvent in a
solution is always lower than
the vapor pressure of the pure
solvent
 Freezing point – temperature at
which the vapor pressures off the
solid phases are the same
o Freezing point depression – 4. Conductivity
the freezing points of solutions - A solution’s conductivity quality depends
are lower than that of the pure principally on the number of respective charges
solvent. It is directly of the ions present
proportional to the molality of - A measure of how well electricity passes through
the solute a solution
 Boiling point – temperature at which - expressed as ohms-1 or mho
the vapor pressure of the solvent Resistivity – reciprocal of conductivity
o Measure of a substance’s resistance to the
reaches one atmosphere
passage of electrical current
o Boiling point elevation – the o The primary application in the clinical
boiling point of solutions are laboratory is for assessing the purity of
higher than that of the pure water
solvent. It is directly
o Expressed as ohms
proportional to the molality of
5. Buffers
solute
- weak acids or bases and their related
 Osmotic pressure – the pressure salts that, as a result of their dissociation
opposing osmosis when a solvent flows characteristics, minimize changes in the
through a semipermeable membrane to hydrogen ion concentration
establish equilibrium between - Hydrogen ion concentration
compartments of differing is often expressed as pH
concentration - pH represents the negative or inverse log
 For a given solute-solvent mass ratio, of the hydrogen ion concentration
all colligative properties are inversely
proportional to solute molar mass
 All of the properties are only colligative
in the dilute limit—at higher
concentrations, the freezing point
depression, boiling point elevation,
vapor pressure elevation or depression,
and osmotic pressure are all dependent - Unlike a strong acid or base, which
on the chemical nature of the solvent dissociates almost completely, the
and the solute dissociation constant for a weak acid or
3. Redox potential – measure of the ability of a base solution tends to be very small,
solution to accept or donate electrons meaning little dissociation occurs
Reducing agents – substances that donate
electrons (lose electrons, oxidized/increases
in oxidation number)
Oxidizing agents – substances that accept
electrons (gain electrons, reduced/decreases in
oxidation number)

Ionic strength – important aspect of buffers,
particularly in separation techniques
 The ionic strength of a solution is a measure of
the concentration of ions in that solution
 Ionic compounds, when dissolved in water,
dissociate into ions. The total electrolyte
concentration in solution will affect important
properties such as the dissociation constant or
the solubility of different salts
 One of the main characteristics of a solution
with dissolved ions is the ionic strength.
 Can be molar (mol/L solution) or molal (mol/kg
solvent) and to avoid confusion, the units should
be stated explicitly
Instrumentation
- All about analytical techniques, methods,
instruments used in cc. In clinical laboratory, there
is a continuous need for quantitative techniques of
measurement.
- Instrumentation has become miniaturized which
enable the development of point of care of device.
- Technologically, sophisticated, automated anlayzers
are still based on the traditional approaches and
technologies.
- Employed photometer in special photometry, ions
selective, electrodes electrophoresis, nephilometry.
- Analytical techniques and instrumentation provides
us foundation for all measurements made in modern
clinical laboratory
Analytical techniques and Instrumentations
 Four basic categories of most measurement
techniques
- Spectrometry – spectrophotometry, atomic
absorption, and mass spectrophotometry.
- Luminiscence – fluorescence, chemiluminescence
and nephelometry.
- Eletroanalytical methods – electrophoresis,
potentiometry, and amperometry.
- Chromatography – gas, liquid and thin layer
techniques.
Photometry
- Instruments that measure electromagnetic radiation have
several concepts and components in common.
- Most frequently used: photometry or specifically
absorbance or reflectance spectrophotometry.
- Photometry employs color and color variation to
determine the concentration of various substances.
- A photometric components is employed in many of the
automated analyzers and any person during clinical lab
techniques should understand the principles of
photometry.
- Photometry is the measurement of the luminous
intensity of light, or the amount of luminous light falling
on a surface from a light source.
- Photometric instruments measure this intensity of light
without consideration of wavelength.
- Spectrophotometry is the measurements of the intensity
of light at selected wavelengths.
- In absorbance spectrophotometry the concentration of
an unknown sample is determined by measuring its
absorption of light at a particular wavelength and
comparing it with the absorption of light by known
standard solutions measured at the same time with the
same wavelength
- The intensity of color is directly proportional to the
concentration of the substance present.
Standard solution – it is a solution containing known
concentration of the same substance to be determined.
- The use of spectrophotometry, or colorimetry as a means
if quantitative measurement depends primarily on two
factors: the color itself and the intensity of the color.
- Any substance to be measured by spectrophotometry
must be naturally colored o must be capable of being
colored.
- An example of a substance that is colored to begin with
is hemglobin. Sugar (glucose) that is not colored to
begin with but is capable of being colored by certain
reaction and reagents.
- When spectophotometry is used as a method for
quantitative measurement, the unknown colored
substance is compared with a similar substance known
strength (a standard solution)
- In absorbance spectrophotometry the absorbance units
or values for serveral different concentrations of a
standard solution are determined by spectrophotometry
and are plotted on graph paper.
- The resulting graph is known as a standard calibration
curve or Beer’s law plot.
- Unknown specimens can then be read in the
spectrophotometer and by use of their absorbance
values, their concentrations determined from the
calibration curve.

NATURE OF LIGHT
- Light is a type of radiant energy and it travels in the
form of waves. The distance between waves is the
wavelength of light.
- Light is used to describe radiant energy with
wavelength visible to the human eye or with
wavelength bordering those visible to the human
eye.
- Electromagnetic radiation includes a spectrum of
energy, from short wavelength, highly energetic
gamma rays and X-rays on the left side to
wavelengths of radio frequencies on the right side
of the spectrum.
- Visible ligt passes between these frequencies, with
the color violet at the 400 nm wavelenth and red at
the 700 nm wavelength.
- The human eye responds to radiant energy, or light,
with wavelenth between 380 and 750 nm
- A nanometer is 1 X 10-9m
- With modern pgotometric appaatues, shorter
(ultraviolet) or longer (infrared) wavelenths can be
measured.
- Modern instruments isolate a narrow wavelenth
range of the spectrum for measurements.
- Most instruments use filters (photometers) or prisms
or gratings (spectrometers) to select or isolate a
narrow range of the incident wavelength.
- The wavelength of light determines the color of the
light seen by the human eye. Every eye color that is
seen is light of a particular wavelength.
- A combination, or mixture of light evergy of different
wavelength is known as daylight or white light. When
light is passed through a filter, a prism, or a
diffraction grating, it can be broken into a spectrum
of visible color ranging from violet to red.
- The color of light seen in the visible spectrum
depends on the wavelength that is not absorbed. When
light is not absorbed, it is transmitted.
- A colored solution has color because of its physical
properties which result in its absorbing certain
wavelengths and tansmitting others.
- When white light is passed through a solution, part of the
light is absorbed and the remaining light is transmitted.
- Meron na a-absorb, meron din na t-transmit
ABSORBANCE AND TRASNMITTANCE OF LIGHT: BEER’S LAW
- Measurements by spectrophotometry is based on the
reaction between the substance to be measured and the
reagent, chemical, used to produce color.
- The amount of color produced in a reaction between the
substance to be measured and the reagent depends on the
concentration of the substance.
- Therefore, the intensity of the color is proportional to the
concentration of the subsatnce.
- Beer’s law states that the concentraion of a substance is
directly proportional to the amount of light absorbed or
inversely proportional to the logarithm if the
transmitted light.
- As the law states, the depth at which the color is
determined must be constant. The depth of the solution
is regulated by the cuvette or container used to hold it.
Beer’s law is the basis for use of photometry in
quantitative measurement.
Any increase in the concentration of the color producing
substance will increase the amount of color seen.
EXPRESSION OF LIGHT TRASNMITTED OR ABSORBED
- There are two common methods of expressing the amount
of light transmitted or light absorbed by the solution.
(another term for absorbed light is optical density, OD)
- The units used to express the readings obtained by the
electronic measuring device are either absorbance (A)
units or percent transmittance (%T).
- Absorbance is an expression of the amount of light
absorbed by a solution.
- Absorbance values are directly proportional to the
concentration of the solution and can be plotted on linear
graph paper to give a straight line.
- Percent transmittance is the amount of light that passes
through a colored solution compared with the amount of
light that passes through a blank or standard solution.
- The blank solution contains all the reagents used in the
procedure, but it does not contain the unknown substance
being measured.
- Blank Solution – to correct the color distribution of the
reagent.
- Percent transmittance varies from 0 to 100 (%T), with
 equal divisions on viewing scale.
- As the concentration of the colored solution
increases, the amount of light absorbed increases,
and the percentage of light transmitted decreases.
- The transmitted light does not decrease in direct
proportion to the concentration or color intensity of
the solution being measured.
- Percent transmittance readings plotted against
concentration will not give a straight line on linear
graph paper.
- A logarithmic relationship exists between %T and
concentration, so when %T is plotted against
concentration, a semi-logarithmic graph paper is
used to obtain a straight line.
PREPARATION AND USE OF A STANDARD CURVE
- In research laboratories or in the preparation of a
new or special procedure, it may be necessary to
prepare a standard curve manually.
- Familiarity with the principles of how a standard
curve is constructed and use.
- Semi-logarithmic graph paper is used to plot %T
readings form the photometer because a logarithmic
relationship exists between percent and
concentration.
- Horizontal axis of the semi-logarithmic graph paper
is a linear scale and the vertical axis is a logarithmic
scale
TYPES OF GRAPH PAPER
- The concentrations of the standard solutions are plotted
on the horizontal axis (x). The transmittance or
absorbance readings from the photometer are plotted
along the vertical axis (y).
- When percentages are plotted against concentrations on
the semi- logarithmic paper, the porportional relationship
is direct and the necessary straight line graph is obtained
when the individual standard points are connected.
- The criteria on good standard curve
o The line is straight
o The line connects all points
o The line goes through the origin, or intersect of
the two axes.
- The origin of the graph paper is the point on the vertical
and horizontal axes where there is %T and zero
concentration.
- Linear graph paper can also be used to plot absorbance
readings because absorbance of wavelengths of light is
directly proportional to the concentration of the colored
solution being read.
o The readings must first be converted to logaritmic
values plotted on the vertical axis.
INSTRUMENTS USED IN SPECTROPHOTOMETRY
- Spectrophotometry is the basis for many of the
instruments used in clinical chemistry.
- The primary reasons for this are ease of measurement,
satisfactory, accuracy and precision, and the suitability
of spectrophotometric techniques to use in automated
instruments.
- In this section, spectrophotometers and colorimeters are
members of this class.
- Most of the instruments used in photometry have some
means of isolating a narrow wavelength, or range of the
color spectrum for measurements.
- Instruments using filters are referred to as filter
photometers, and those using prisms or diffraction
grating are called spectrophotometers or
photoelectric colorimeters.
- Photometers utilize an electron device to compare
the actual color intensities of the solutions
measured.
- Spectrophotometer is really 2 components in a
single case, a spectrophotometer, a device for
producing light of a specific wavelength, the
monochomator; and a photometer a device for Hold the spectrophotometers on the top only.
measuring light intensity. - Barrier-layer cell
o Least expensive
PARTS OF SPECTROPHOTOMETERS
o Composed of light-sensitive materials such as
- Light source: the light source supplies the
selenium on iron plate with transparent layer
radiant energy used to analyze the sample. The
of silver.
most common source of light for work in the
o When exposed ti light, electrons in the light
visible or near-infrared region is the
sensitive material are excited and are
incandescent tungsten or tungsten-iodine
released into the highly conductive silver.
lamp. Most emitted radiant energy is near-
o No need for power source.
infrared. The most common lamps for UV work
are deuterium-discharge & mercury oil lamps.
- Photoemissive/phototube
- The wavelength isolator: a system of isolating
o Has photosensitive material that gives off
a desired wavelength and excluding others is
electrons when light energy strikes it.
called a monochromator.
o Composed of positively charged anode and
- Some filters are very simple, composed of one
negatively charged cathode enclosed in a
or two pieces of colored glass. The filter must
glass.
transmit a color that a solution can absorb.
o The cathode emits electrons jump over to the
- One common instruments amploys a diffraction
positively charged anode where they are
grating with a special plate and slit to reduce
collected and return through an external
the spectrum to the desired wavelength. The
measurable amount.
grating consists of a highly polished surface with
o Required an external source of energy.
numerous lines that break up white light into
spectrum.
- Photomultiplier
- By moving the spectrum behind a slit (the light
o Emits electrons proportionally to initial light
source must be movable). Only one particular
absrobed, convert radiant ebergy to
portion of the spectrum is allowed to pass
electrical energy.
through the narrow slit. The paricular band of
o Must be shielded from stray light.
light, or wavelength transmitted through the slit
o Ampplified the initial signal received
is indicated on a viewing scale on the machine.
o Has a very rapid response time
o Most senisitive to low levels of light.
- Photodiode array
o A type of photodetector that has
photoiodides ina line or arrangement
allowing it to respons to a specific
wavelength resulting in complete UV/visible
- The cuvette holds the sample to be analyzed in
spectrum analysis.
the path of the energy.
o Produces a spectral absorbance curve.
- Cuvettes
o are made from plastic, glass or quartz.
- The amount of light transmitted by the solution in
These are acceptable in visible work.
the cuvette is measured by the photoelectric cell,
o There are inexpensive plastic cuvettes
that may be suitable for some UV work. producing electrons in proportion to the amount of
light hitting it. The electrons are passed on to a
o Analytical cell or sample cell.
galvanometer, where they are measured.
- The galvanometer records the amount of current
that it receives from the photoelectric cell on a
special viewing scale (read-out device) on the
 spectrophotometer. The results are reported in
terms of T% (absorbance).
Quality Control Tests
- Wavelength accuracy is ensured when the
wavelength indicated on the control dial is the
actal wavelength of light passed by the
monochromator.
- This is checked with standard absorbing solution
or filters with maximum absorbance of known
wavelength.
- Wavelength calibration can be tested using rare-
earth glass filters (didymium), or holmium oxide
glass filters or a stable chromogen solution.
- Photoelectric accuracy can be checked by
reading standard solutions of potassium
dichromate or potassium nitrate.
- Stray light refers to any wave lengths outside
tha band transmitted by the monochromator.
The most common causes of stray light are:
1. Reflections of light from stcratches on
optical surfaces or form dust particles
anywhere in the light path.
2. Higher-order spectra produced by diffraction
gratings
- Stray light is detected by using cutoff filters.
Instrumentation
Atomic Absorption Spectrophotometer
 Principle – Ground-state atoms absorb light at defined
wavelengths
- Used to measure concentration by detecting the
absorption of electromagnetic radiation by atoms
rather than by molecules
- Line spectrum refers to the wavelength at which an
atom absorbs light, each metal exhibits a specific line
spectrum
- The sample is atomized in a flame where the atoms
of the metal to be quantified are maintained at ground
state
- Then a beam of light from a hollow-cathode lamp
(HCL) is passed through a chopper to the flame
- The ground state atoms in the flame absorb the same
wavelengths of light from the HCL as the atoms emit
when excited
- The light not absorbed by the atoms is measured as a
decrease in light intensity by the detector
- The detector (photomultiplier tube) will selectively
read the pulsed light from the chopper that passes
through the flame and will not detect
any light emitted by the excited atoms when they
return to ground state
- The difference in the amount of light leaving the
HCL, and the amount of light measured by the
detector is indirectly proportional to the
concentration of the metal analyte in the sample
(A is directly proportional to the %T)
 Components
- Hollow-cathode lamp (light source) – consists
of an evacuated gas-tight chamber containing an
anode, a cylindrical cathode and an inert gas, such
as helium or argon
- Chopper – used to break the steady light into
pulsating light. It is a rotating wheel placed
between the flame and the source
- Burner head for flame – sample cell
- Monochromator
- Detector
- Readout device
 Applied voltage causes ionization of the gas, and these
excited ions are attracted to the cathode, where they
collide with the metal coating on the cathode,
knocking off atoms and causing atomic electrons to
 become excited
 When the electrons of the metal atoms from the
cathode return to the ground state, the
characteristic light energy of the metal is
emitted
 Vaporized metal atoms from the sample can be
found in the flame. The flame serves as the
sample cuvet in this instrument and bring the
analyte to its ground state
 The light produced in the HCL passes through a
chopper (to avoid errors caused by
measurement of light emitted by the sample)
and then to the flame, and the light is absorbed
by the metal in the sample. The light not
absorbed will be read by the photomultiplier
tube
 A flameless system employs a carbon rod
(graphite furnace), tantalum, or platinum to
hold the sample being analyzed. The atomized
sample then absorbs the light energy from HCL.
This technique is more sensitive than the flame
method.
 In assaying analyte with a single-beam atomic
absorption spectrophotometer, the instrument
is actually measuring the intensity of the beam
from the hollow cathode lamp after it has
passed through the analyte-containing flame
 It is accurate, precise and specific, one
disadvantage is the inability of the flame to
dissociate samples into free atoms
 Another possible theorem is the ionization of
atoms following dissociation by the flame,
which can be decreased by reducing the flame
temperature
- Matrix interference, due to the enhancement of
light absorption by atoms in organic solvents or
formation of solid droplets as the solvent
evaporates in the flame, can be another source
of error
- This interference may be overcome by
pretreatment of the sample by extraction
TURBIDIMETRY
1. Measures light blocked as a decrease in the light
transmitted through the solution, dependent on particle
size and concentration.
2. Uses a spectrophotometer for measurement, and is
limited by the photometric accuracy and sensitivity of the
instrument.
3. The problems inherent in turbidimetry
A. variation in particle size of samples
B. variation in particle size of standards
C. rate of aggregation or settling of particles
NEPHELOMETRY
1. The measurement of light scattered by a particulate
solution.
2. Generally, scattered light is measured at an angle to
the incident light when small particles are involved; for
large molecules, forward light scatter can be measured.
3. The amount of scatter is directly proportional to the
number and size of particles present in the solution.
4. The sensitivity of nephelometry depends on the
absence of background scatter from scratched cuvets and
particulate matter in reagents.
MOLECULAR EMISSION SPECTROSCOPY
I. Types of luminiscence where excitation does
requires absorption of radiant energy.
A. Fluorescence is the emission of light by a
substance that has absorbed light or other
electromagnetic radiation.
It is a form of luminiscence, in most cases, the emitted
light has a longer wavelength, and therefore lower energy
than the absorbed radiation.
I. Fluorometry is defined as the measurement of the
emitted fluorescence light.
The excitation light is absorbed by the atoms of the
analyte in solution, which causes the electrons to move
to higher energy orbitals.
Upon return to ground state, light is emitted from the
fluorescing analyte and that light passes through a
secondary filter.
The secondary filter and the detector are placed at a right
angle to the light source to prevent incident light from
being measured by the detector.
Whereas fluorometers use filters, spectro-fluorometers
use prisms or diffraction gratings as monochromators.
2. Advantages: Fluorometry is about 1000 times more
sensitive than absorption techniques and has increased
specificity because optimal wavelengths are chosen both
for absorption (excitation) and for monitoring emitted
fluorescence.
3. Limitation: Changes from the established protocol that
affect pH, temperature, and solvent quality; self-
absorption: quenching.
Fluorometers are designed so that the path of exciting
light is at a right angle to the path of the emitted
light and this design prevents excitation light from
reaching the detector.
Requires primary and secondary monochromator.
Primary advantage of performing fluorometric over
absorption spectrosopic methods of analysis is
increased specificity and increased sensitivity.
b. Phosphorescence is the emission of light
produced by certain substances after they absorb
energy.
It is similar to fluorescence except that the
time delay is longer (greater than 10−4 sec) between
absorption of radiant energy and release of energy as
photons of light.
2. Types of luminiscence where excitation does not
require absorption of radiant energy
a. Chemiluminiscence is the process where the
chemical energy of a reaction produces excited
atoms and upon electron return to ground state,
photons of light are emitted.
Advantage: Has excellent sensitivity and dynamic
range
Ruthenium is the substance used to generate light
signal.
Source lamp is not needed in chemiluminiscent
immunoassay, has monochromator, photodetector
and wash solution.
b. Bioluminiscence is the process where an enzyme-
catalyzed chemical reaction produces light
emission.
For ex., this may occur in the presence of the enzyme
luciferase because of oxidation of the substrate
luciferin.
Luminometer is a generic term for the type of
instrument that is used to measure
chemiluminiscence and bioluminiscence.
CHROMATOGRAPHY
1. A technique where solutes in a sample are
separated for identification based on physical
differences that allow their differential distribution
between a mobile phase and a stationary phase.
2. Mobile phase may be an inert gas or liquid.
3. Stationary phase may be silica gel bound to the
surface of a glass plate or plastic sheet; may be silica
or a polymer that is coated or bonded within a
column.
THIN-LAYER CHROMATOGRAPHY (TLC)
It is a type of planar chromatography. The stationary
phase may be silica gel that is coated onto a solid
surface such as a glass plate or plastic sheet.
The mobile phase is a solvent, where solvent polarity
should be just enough to achieve clear separation of
the solutes in the sample. It is used clinically for urine
drug screening.
The mobile phase moves through the stationary
phase by absorption and capillary action.
The solute components move at different rates because
of solubility in the mobile phase and electrostatic forces
of the stationary phase that retard solute movement.
These two phase work together to provide solute
resolution and separation.
a. Solute will stay with the solvent front if solvent is too
polar for the solute.
b. Solute will remain at origin if solvent is insufficiently
polar.
Basic steps in performing TLC include
* sample extraction using a liquid-liquid or column
technique
* concentration of the extracted sample;
* sample application by spotting onto the silica gel plate
* development of the solute in the sample using stationary
and mobile phases
* solute detection using chromogenic sprays, UV light,
fluorescence, and heat and
* interpretation of chromatographic results utilizing Rf
values of solutes in comparison to aqueous standards.
In TLC, the Rf is the distance the solute migrates
divided by the distance the solvent migrates.
Rf values are affected by chamber saturation,
temperature, humidity, and composition of the solvent.
GAS-LIQUID CHROMATOGRAPHY (GLC)
1. Components include
* a carrier gas with a flow-control device to regulate the
gas flow
* a heated injector
* chromatographic column to separate the solutes
* heated column oven
* detector
*computer to process the data and control the operation
of the system
2. Gas-liquid chromatography is a technique used to
separate volatile solutes.
a. The sample is injected into the injector component of
the instrument where the sample is vaporized because
the injector is maintained approximately 50°C higher
than the column temperature.
b. An inert carrier gas (mobile phase) carries the
vaporized sample into the column. Carrier gases
commonly used include hydrogen, helium, nitrogen and
argon.
The carrier gas flow rate is critical to maintaining column
efficiency and reproducibility of elution times.
The elution order of volatiles is usually based upon the
boiling point.
c. The types of columns (stationary phase) used are
designated as packed or capillary.
When the volatile solutes carried by the gas over the
stationary phase of the column are eluted, the column
effluent is introduced to the detector.
The solutes are introduced to the detector in the order
that each was eluted.
d. The detector produces a signal for identification and
quantification of the solutes.
Commonly used detectors include flame ionization,
thermal conductivity, electron capture and mass
spectrometer.
e. Separation of solutes is a function of the relative
differences between the vapor pressure of the
solutes and the interactions of the solutes with the
stationary column.
The more volatile a solute, the faster it will elute
from the column; the less interaction of the solute
with the column, the faster it will elute.
f. Identification of a solute is based on its retention
time, and quantification is based on peak size,
where the amount of solute present is proportional
to the size of the peak (area of height of the sample
peak, is compared to known standard).
In addition, GLC
a. separation depends on the sample and solubility in
the liquid layer of the stationary phase
b. stationary phase is a liquid layer adsorbed on the
column packing.
HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
(HPLC)
1. HPLC components include
* sample-introduction system
* solvent reservoirs (solvent-delivery system)
* one or more pumps to propel the solvent(s)
* injector
* chromatographic column
* detector
* computer to process data & control the operation
of the system
2. HPLC is a type of liquid chromatography where the
mobile phase is a liquid that is passed over the
stationary phase of the column.
The separation of solutes in a sample is governed by
the selective distribution of the solutes between the
mobile and the stationary phase.
a. Solvents commonly used for the mobile phase
include acetonitrite, methanol, ethanol, isopropanol
and water.
1. Isocratic elution: strength of solvent remains
constant during separation
2. Gradient elution: strength of solvent continually
increases (%/min) during separation
b. Stationary phase is an organic material covalently
bonded to silica that may be polar or nonpolar in
composition.
1. Normal-phase liquid chromatography: polar
stationary phase and nonpolar mobile phase
2. Reversed-phase liquid chromatography:
nonpolar stationary and polar mobile phase
3. The solvent-delivery system utilizes a solvent
reservoir from which the pump can push the mobile
phase through the column.
The sample is introduced through a loop injector.
A pre-column and guard column function to maintain the
integrity of the column and are positioned prior to the
sample reaching the main column.
The column, which functions as the stationary phase,
generally operates at room temperature.
The effluent from the column passes to a detector
system.
The solutes are introduced to the detector in the order
that each was eluted.
4.The detector produces a signal for identification and
quantification of the solutes.
Commonly used detectors include spectrophotometer,
photodiode array, fluorometer, electrochemical (glassy
carbon electrode)
It separates solutes in a sample based on the sign and
magnitude of the ionic charge.
MASS SPECTROMETRY
1. A mass spectrometer is an instrument that uses the
principle of charged particles moving through a magnetic
or electric field with ions being separated from other
charged particles according to their mass-to-charge
ratios (parameter used to identify a compound)
In this system, electrons bombard a sample, ionizing the
compound into fragment ions, which are separated by
their mass-to-charge ratios.
The mass spectrum produced is unique for a compound
(identification), and the number of ions produced relates
proportionally to concentration (quantification).
2. Mass spectrometry is a high-quality technique for
identifying drugs or drug metabolites, amino acid
composition of proteins, and steroids.
In addition, mass spectrometry has applications in the
field of proteomics.
The eluate gas from a gas chromatograph may be
introduced into a mass spectrometer that functions as
the detector system, or the liquid eluate may be
introduced from a high-liquid chromatograph.
3. Instrumentation: Mass spectrometer components
include
a. Ion source: samples enter the ion source and are
bombarded by the ionization beam.
When the sample is in gas form and introduced from a gas
chromatograph, the ion source may be electron or
chemical ionization.
Other types, such as electrospray ionization and sonic
spray ionization, may be used when a high-performance
liquid chromatograph is used in conjunction with
spectrometer.
b. Vacuum system: prevents the collision of ions with
other molecules (fragments) when electronic or magnetic
separation is occurring.
c. Analyzer: Beam-type and trapping-type
1. Beam-type is a destructive process, where ions
pass through the analyzer one time and then strike the
detector.
2. Quadropole is a beam-type analyzer, where
mass-to charge ratios are scanned during a prescribed
time period to form a mass spectrum.
d. Detector usually detects ions using electron
multipliers, such as discrete dynode and continuous
dynode electron multipliers.
e. Computer and software convert the detector's
signal to a digital form.
Sample identification is achieved because
each compound produces a unique spectrum, which
is analyzed by a database for matching to a
computerized reference library.
4. To further improve selectivity and sensitivity, a
system known as tandem mass spectrometers can be
employed, where a gas chromatograph is connected
to two mass spectrometers (GC MS/MS) or
(HPLC/MS/MS).
In these systems, ions of a specific mass-to-
charge ratio are allowed to continue to the second
mass spectrometer where additional fragmentation
occurs and final analysis is done.
ELECTROCHEMISTRY
1. When chemical energy is converted to an electrical
current (a flow of electrons) in a galvanic cell, the
term electrochemistry is used.
2. Electrochemical reactions are characterized by a
loss of electrons (oxidation) at the positive pole
(anode) and a simultaneous gain of electrons
(reduction) at the negative pole (cathode).
3. The galvanic cell is made up of two parts called
half-cells, each containing a metal in a solution of
one of its salts.
4. These methods involve the measurement of
electrical signals associated with chemical systems
that are within an electrochemical cell.
5. In the clinical lab, electroanalytical methods are
used to measure ions, drugs, hormones, metals and
gases.
6. Methods are available for the rapid analysis of
analytes present in relatively high concentrations in
blood and urine, such as blood electrolytes (Na+, K+,
Cl-, HCO3-), and other analytes present in very low
concentrations, such as heavy metals and drug
metabolites.
1. Many types of electrochemical analyses are used
including potentiometry, amperometry,
coulometry and polarography.
2. The two basic electrochemical cells involved in
these analyses are galvanic and electrolytic cells.
Polarography, analytical procedure measuring
concentration of a substance based on the amount of
current flow resulting from ion transfer when
substance go into chemical reaction.
It employs an electrochemical cell.
a. Gradually increasing the voltage applied between
two electrodes of the cell in contact with a solution
containing the analyte.
b. Current measured: voltage change versus plotted to
produce a polarogram. .
c. Voltage at which sharp rise in current occurs
characteristic of the electrochemical reaction involved.
d. Amount of increase in current (i.e. wave height)
proportional to the concentration of analyte.
2. Anodic stripping voltammetry is based on polarography
a. Negative potential applied to one of the electrodes.
b. Trace metal ions in the solution reduced and plated
onto anodic electrode; preconcentrating step.
c. Plated electrode used as anode in polarographic cell;
metal stripped off anode.
d. Current flow during stripping provides polarogram that
identifies and quantifies the analyte being measured
(trace metals).
e. Used to assay heavy metals such as lead in blood.
1. Potentiometry is a technique used to determine the
concentration of a substance in solution employing an
electrochemical cell that consists of two half-cells,
where the potential difference between an indicator
electrode and a reference electrode is measured.
a. Half cell, also called an electrode, composed of single
metallic conductor surrounded by solution of electrolyte.
b. In an electrolytic cell, the half cell where reduction
takes place is a cathode.
c. Two different half-cells connected to make complete
circuit; current flows because of potential difference
between two electrodes.
d. Salt bridge connection between two metallic
conductors and between two electrolyte solutions.
e. Comparison made between the voltage of one half-cell
connected to another half-cell.
f. Half-cell potentials compared to potential generated
by standard electrode
g. Universally accepted standard half-cell is the standard
hydrogen electrode, arbitrally assigned a potential E° of
0.000 volt.
h. Desirable to use one half-cell (reference electrode)
with known and constant potential, not sensitive to
composition of material to be analyzed.
i. Calomel electrode type of reference electrode,
consisting of mercury covered by a layer of mercurous
chloride in contact with saturated solution of potassium
chloride
j. Other half-cell (indicator electrode) selected on basis
of change in its potential with change in concentration of
analyte to be measured.
k. Silver-silver chloride (Ag/AgCl) electrode; common
type of reference electrode.
2. A pH/blood gas analyzer employs a pH-sensitive glass
electrode for measuring blood pH, and it employs PCO₂
and PO₂ electrodes for measuring gases in blood
For measuring pH, the pH electrode is a functioning glass
electrode that is dependent on properties of pH-
sensitive glass
When a pH-sensitive glass electrode is not actively
in use, it should be kept in a medium recommended
by the manufacturer
To calibrate the pH electrode, it is necessary that
calibrating gases, two buffer solutions of known pH
be used
a. Glass electrode made by sealing thin piece of pH-
sensitive glass at the end of glass tubing and filling
tube with solution of hydrochloric acid saturated with
silver chloride
b. Silver wire immersed in tube's solution with one
end extending outside the tube for external
connection, silver-silver chloride reference electrode
sealed within tube with pH-sensitive glass tip
c. pH sensitive glass must be saturated with water.
Surface of the glass develops a hydrated lattice,
allowing exchange of alkaline metal ions in the
lattice for hydrogen ions in the test solution.
A potential is created between the inside and the
outside of the electrode, and that potential is
measured
d. Glass electrode calibrated by comparison with two
primary standard buffers of known pH
e. Because pH readings are temperature sensitive,
the calibration must be carried out at a constant
temperature of 37°C
3. In a pH/blood gas analyzer, the PCO, electrode for
measuring the partial pressure of carbon dioxide
(PCO₂) in blood is actually a pH electrode immersed
in a bicarbonate solution
a. The bicarbonate solution is separated from the
sample by a membrane that is permeable in gaseous
CO₂ but not to ionized substance such as H+ ions
b. When CO₂ from the sample diffuses across the
membrane, it dissolves, forming carbonic acid and
thus lowering the pH.
The measurement of CO₂ in blood by means of PCO2
electrode is dependent on the change in pH because
of increased carbonic acid in the electrolyte
surrounding the electrodes.
The pH is inversely proportional to the log of the
PCO₂. Hence the scale of the meter can be calibrated
directly in terms of PCO₂
4. The ion-exchange electrode is a type of
potentiometric, ion-selective electrode
a. The ion-selective electrode universally used is the
pH
electrode
Consists of liquid ion-exchange membrane
made of inert solvent and ion-selective neutral
carrier material
b. Collodion membrane may be used to separate
membrane solution from sample solution
c. K analysis: antibiotic valinomycin, because of its
ability to bind K+ used as a neutral carrier for K+ selective
membrane
d. NH4+ analysis: antibotics nonactin and monactin used
in combination as neutral carrier for NH4+--selective
membrane
5. Sodium analysis: ion-selective electrodes based on
principle of potentiometry
a. Unlike glass membrane electrodes with selective
capability
b. Constructed from glass that consists of silicon dioxide,
sodium oxide and aluminum oxide
c. lon-selective electrode analysis of sodium
* uses a glass membrane
* error occur from protein buildup on the membrane
* principle is based on potentiometry
* membrane not coated with valinomycin
Amperometry: Electrochemical technique that measures
the amount of current (measures the increase in
conductivity) produced through the oxidation or
reduction of the substance to be measured at an
electrode held at a fixed potential.
1. In a pH/blood analyzer, the electrode for
measuring the partial pressure of oxygen (PO₂) in
the blood is an electrochemical cell consisting of
a platinum cathode and a Ag/AgCI anode
connected to an external voltage source.
2. The cathode and anode are immersed in the
buffer. A polyropylene membrane selectively
permeable to gases separates the electrodes and
buffer from the sample.
3. When there is no oxygen diffusing into the buffer,
there is practically no current flowing between
the cathode and the anode because they are
polarized.
4. When oxygen diffuses into the buffer from a
sample, it is reduced from the platinum cathode.
5. The electrons necessary for this reduction are
produced at the anode. Hence, aa current flows;
the current is directly proportional to the PO2, in
the sample.
Coulometry
1. A chloride coulometry employs a coulometric
system based on Faraday's law, which states that
in an electrochemical system, the number of
equivalent weight of a reactant oxidized or
reduced is directly proportional to the quantity of
electricity used in the reaction.
The quantity of electricity is measured in coulombs.
Coulomb is the unit of electrical quantity; I
coulomb of electricity flowing per minute
constitutes a current of I ampere.
 2. If the current is constant, the number of
equivalent weights of reactant oxidized or
reduced depends only on the duration of the
current.
3. In the chloride coulometer, the
electrochemical reaction is the generation of
Ag+ ions by the passage of a direct current
across a pair of silver electrodes immersed in
a conducting solution containing the sample
to be assayed for chloride.
As the Ag+ ions are generated, they are
immediately removed from solution by
combining with chloride to form insoluble
silver chloride.
When all the chloride is precipitated; further
generation of Ag+ ions causes an increase in
conductivity of the solution.
4. The endpoint of the titration is indicated by
the increase in conductivity of the solution.
COULOMETRIC CHLORIDOMETEDS AND ANODIC
STRIPPING VOLTAMMETRY
Chloride ISEs have largely replaced coulometric
titrations for the determination of chloride in body
fluids.
Anodic stripping voltametry was widely used for the
analysis of lead and is best measured by
electrothermal (graphic furnace), atomic absorption
spectroscopy or, preferably ICP MS)
ELECTROPHORESIS
1. Used clinically to separate and identify proteins,
including serum, urine and cerebrospinal fluid (CSF)
proteins, lipoproteins, isoenzymes and so on.
2. Electrophoresis is defined as the movement of
charged molecules in a medium when an electric field
is applied.
3. Zone electrophoresis is defined as the movement
of charged molecules in a porous supporting medium
where the molecules separate as distinct zones.
4. Support medium provides a matrix that allows
molecules to separate (e.g. agarose gel, starch gel,
polyacrylamide gel, and cellulose acetate
membranes.
5. Movement of charged particles through a medium
depends on the nature of the particle, including net
charge, size and shape, the character of the buffer
and supporting medium, temperature and the
intensity of the electric field.
a. Nature of the charged particle. Proteins are
amphoteric and may be charged positively or
negatively depending on the pH of the buffer
solution.
b. The pH at which negative and positive charges are
equal on a protein is the protein's isoelectric point.
6. Buffer solutions of pH 8.6 are generally used for serum
protein electrophoresis. Using agarose gel or cellulose
acetate at this alkaline pH, serum proteins take on a net
negative charge and will migrate toward the anode (+).
Albumin migrates the fastest toward the anode
and the gamma globulins remain closer to the cathode (-
).
7.Visualizing the separated analyte: Following
eletrophoresis, treat the support medium with
colorimetric stains or fluorescent chemicals.
Amido black B, Ponceau S and Coomassie brilliant blue
stains are used for visualizing serum proteins.
Silver nitrate is used for CSF proteins, fat red 7B and oil
red O are used for lipoproteins and nitrotetrazolium blue
is used for lactate dehydro-genase.
8. Detection and quantification of the separated protein
is accomplished using densitometer.
9. Commonly encountered problems in electrophoresis:
a. Holes in staining pattern: Analyte present in too
high a concentration
b. Very slow migration: Voltage too low
c. Sample precipitates in support: pH too high or
low; excessive heat production
10. Isoelectric focusing is a type of zone electrophoresis
in which protein separation is based on the isoelectric
point (pl) of the proteins.
This method utilizes polyacrylamide or agarose gel
containing a pH gradient formed by ampholytes in the
medium.
When exposed to an electric field, the ampholytes
migrate based on their pl to their respective positions in
the gradient. In turn, the serum proteins will migrate in
the gel to the position where the gel's pH equals the pl of
the respective protein.
In turn, the serum proteins will migrate in the gel to the
position where the gel's pH equals the pl of three
respective protein.
11.Capillary electrophoresis is based on electroosmotic
flow (EOF). When an electric field is applied, the flow of
liquid is in the direction of the cathode. Thus, EOF
regulates the speed at which solutes move through the
capillary.
AUTOMATION PARAMETERS/TERMINOLOGY
1. Centrifugal analysis: Centrifugal force moves samples
and reagents into cuvet areas for simultaneous analysis.
2. Discrete analysis: Each sample reaction is
compartmentalized.
This may relate to an analyzer designed to assay
only one analyte (e.g. glucose) or an analyzer capable of
forming multiple tests where the sample and reagents are
in a separate cuvet/reaction vessel for each test.
3. Random access: Able to perform individual tests or
panels, and allows for stat samples to be added to run
ahead of other specimens.
4. Batch analysis: Samples processed as a group.
5. Stand-alone: Instrument from a single discipline
with automated capability.
6. Automated-stand alone: Instrument from a single
discipline with internal automated capability (e.g.
auto-repeat and auto-dilute.)
7. Modular workcell: At least two instruments from a
single discipline with additional internal automated
capability.
8. Multiple platform: Instrument able to perform
tests from at least two disciplines.
9. Integrated modular system: At least two analytical
modules supported by one sample and reagent
processing and delivery system.
10. Pneumatic tube system: Transports specimens
quickly from one location to another.
11. Throughput: Maximum number of tests generated
per hour
12.Turnaround: Amount of time to generate one
result
13. Bar coding: mechanism for patient/sample
identification; used for regent identification by an
instrument.
14. Dead volume: Amount of serum that cannot be
aspirated
15. Carry-over: The contamination of a sample by a
previously aspirated sample
16. Reflex testing: Use of preliminary test results to
determine if additional tests should be ordered or
cancelled on a particular specimen; performed
manually or automated
17. Total laboratory automation: Automated systems
exist for laboratories where samples are received,
centrifuged, distributed to particular instruments
using a conveyor system, and loaded into the
analyzer without operator assistance.
This kind of automation is seen in large
medical center laboratories and commercial
laboratories where the volume of testing is high.
PRINCIPLES OF AUTOMATION
1. Automated instruments use robotics and fluidics to
replicate manual tasks.
2. Specimen handling: Some instruments have level-
sensing probes that detect the amount of serum or
plasma in the tube.
Some systems have a reading device that
allows bar-coded sample tubes to be loaded onto the
instrument.
Although not as common, other instruments
require the operator to manually enter the position
of the patient sample.
3. Reagents
a. Dry reagents can be packaged as lyophilized
powder or tablet form that must be reconstituted
with a buffer or reagent grade water.
Reconstituting of reagents may need to be done
manually and then the reagents placed on an analyzer for
use, or reconstituting the reagents may be part of the
total automation process as employed by Dimension®
analyzer.
b. Dry reagents can be spread over a support material and
assembled into a single-use slide. This technique is
employed by the Vitros® analyzer.
c. Liquid reagents are pipetted by the instrument and
mixed with the sample.
4. Testing Phase
a. Mixing sample and reagents occurs in a vessel called a
cuvet. Some instruments have permanent, nondisposable
cuvets made of quartz glass. Other cuvets are made of
plastic and are disposable.
b. Reaction temperatures and times vary for each
analyte. The most common reaction temperatures are
37°C and 30°C.
c. Kinetic assays: Determination of sample concentration
is based on change in absorbance over time.
d. Endpoint/colorimetric assays: Incubated for a specific
time, absorbance determined, absorbance related to
calibrators for calculation of sample concentration.
e. A spectrophotometer is built within the system to read
absorbances for kinetic and colorimetric assays.
These systems may use a diffraction grating or aa
series of high quality fibers. Some automated analyzers
incorporate fluorometry or nephelometry.
5. Data Management
a. The computer module of most automated instruments
has a data management system that allows analysis of
quality control (QC) materials and assessment of patient
values (e.g. delta check) before releasing patient results.
b. Instruments/laboratory information systems (LISs) also
archive patient results and QC values. These archived
results are stored by the laboratory for various lengths of
time.
POINT-OF-CARE TESTING (POCT)
1. Definition: Performing diagnostic tests outside the
main laboratory and at or near patient care areas.
2. Applications: POCT is designed to provide immediate
laboratory test results for immediate patient assessment
and determination of appropriate treatment.
POCT may be used in neonatal intensive care,
coronary care, intensive care or the emergency
department.
3. Operators
Only waived laboratory tests can be performed
using point-of-care instruments.
Clinical laboratory technicians and clinical
laboratory scientists must operate instruments that
perform complex or high-complexity laboratory tests.
4. Point-of-care (POC) instrument evaluation
All POC instruments must be evaluated in accordance
with the Clinical Laboratory Improvement Amendments of
1988 (CLIA '88).
The values obtained from POC instruments must
correlate with values obtained from larger laboratory
instruments.
Linearity testing, calculation of control ranges,
correlations of sample data and reference ranges
must be done for each instrument
5. Training: All POC instrument operators must be
trained, and training must be documented.
6. Quality control: All effective quality control
systems must be set up for each POC instrument. The
program must use appropriate standards and
controls, statistical analyses, and a proficiency
testing system.
This information must be documented.
LONG QUIZ CC 2021 o Deionization
o Filtration
1. Glassware of choice for dilution and o Reverse osmosis
preparation of solutions that are made to o Distillation
definite volume 8. Which of these is a correct colligative property?
o Erlenmeyer Flask o Boiling point depression
o Volumetric Flask o Density
o Graduated Cylinder o Freezing point elevation
o Automatic pipet o Vapor pressure lowering
9. Mechanism that draws up and dispenses the liquid
2. All of the ff. describes positive displacement
is an integral part of the pipet
micropipette except
o Mohr pipet
o No air cushion
o Volumetric pipet
o With disposable piston
o Automatic pipet
o Piston is permanent part of the
o Serological pipet
pipet
10. Refers to how close a group of measurements are
o For volatile sample
to each other
o Estimate
3. The boiling point of a solution is ______ that
o Precision
of a pure solvent.
o Accuracy
o Lower than
o Significant
o Less than
11. Which of the ff. is material of known composition
o Higher than
available in a highly purified form?
o Equal to
o Control
4. The freeing point of a solution is _____ that
o Technical reagent
of a pure solvent.
o Test analyte
o Higher than
o Standard
o More than
12. When using a micropipette, if you push to the
o Equal to
second stop to fill it you will get
o Lower than
o The correct amount of solution
5. Which of the following terms refer to how
o The little solution
close an analytical value approaches the
o Too much solution
actual value of the substance during the
o Either too much or too little solution
assay?
13. Colligative properties of solutions are those
o Accuracy
properties that depend on
o Reliability
o The number of solute only
o Specificity
o Neither the number and nature of the
o Precision
solute
6. This is used for substances that are
o The nature of solute only
particularly sensitive to light such as
o Both the number and nature of solute
bilirubin.
14. Molality is
o Soft glass
o The moles of solute in 1g of solvent
o Low actinic glass
o The moles of solute in 1g of solution
o Borosilicate glass
o The moles of solute in 1 kg of solvent
o Soda-lime glass
o The moles of solute in 1kg of solution
https://labpedia.net/laboratory-glassware-cleaning-and- 15. What happens to a decimal place in the number
sterilization/ when you are converting from a large unit to a
smaller unit?
7. A method of purifying water where it is made o Converting doesn’t require a placement of
to pass over an insoluble resin polymer that a decimal point
exchange the hydrogen ions for the ionized o The decimal place moves to the left
impurities in the water. The principle is o The decimal place doesn’t move at all
 o The decimal moves to the right
16. The chemical reagent has the highest purity
for laboratory use
o USP
o None of the option is correct
o Technical grade
o Analytical grade
17. Reagent grade Type I water is used in
o Highly sensitive procedures
o Preparation of standards
o All options are correct
o Procedures requiring minimal
interference
18. Pipette with a long, cylindrical tube drawn
out to a tip and is calibrated for blown out
and uniform fractional volume of
measurement is known as
o To deliver pipette
o Volumetric pipette
o To contain pipette
o Serological pipette
19. The molecular weight divided by the valence
is the
o Formula weight
o Equivalent weight
o Standard weight
o Molal weight
20. Best glassware for preparation of reagent
such as 100 mL of 10% sodium tungstate
o 100 mL graduated cylinder
o 100 mL beaker
o 100 mL Erlenmeyer flask
o 100 mL volumetric flask
21. A molar solution contains
o None of the options is correct
o 1g equivalent weight of solute per
liter of solution
o 1g solute to 99g of solvent
o 1g molecular weight (gr.w) of solute
per liter of solution
22. An expression of one amount relative to
another
o Molarity
o Ratio
o Dilution
o Concentration
23. Of a volumetric pipette mark with a ring near
the end, the solution must be
o Drain out
o Rinsed out
o Blown out
o Left in the pipet before rinsing out with
distilled water
24. Which of the ff. does not characterize borosilicate
glass?
o Has a high thermal resistance
o Does not contaminate its content
o Imparts amber/brown color to the glass
o Has a low alkali content
25. Substance that resemble the unknown specimen
and used to measure precision
o Callibrator
o Buffer
o Blank
o Control
26. Following the standard Beer’s Law, given diluted
volume of 5,10 ,20, and 40 standard what will be
the appearance of the graph? (di ko pa ren to
sure)
o A straight line on percent transmission
o A curve line on semilog paper
o A straight line on semilog paper
o A straight line on a linear paper
27. In absorption spectrometry
(mali yung asa quiz, tama na ito)
o Absorbance is directly proportional to
transmittance
o Percent transmittance is directly
proportional to concentration
o Percent transmittance is directly
proportional to the light path length
o Absorbance is directly proportional to
concentration
28. If the ff. reaction is known to obey Beer’s Law,
the X axis is the concentration of the solution and
the Y axis is
o Absorbance
o Percent transmittance
o Peak weight
o Wavelength
29. Which measurement requires a primary and
secondary monochromator?
o Spectrophotometer
o Fluorometer
o Atomic absorption spectrophotometer
o Nephelometer
30. Similar to optical density
o Emission
o Transmittance
o Percent
o Absorbance
31. Which substance is used to generate the light
signal in electrochiluminescence?
 o Ruthenium
o Dioxetane phosphate
o Luminol
o Acridinium
32. Beer’s law states that the concentration of a
substance is
o Concentration are all the same
o Directly proportional to the square
root of a substance
o Immersely proportional to the amount
of concentration
o Directly proportional to the amount
of the light absorbed
33. Reagent grade type III water is used in
o Qualitative test in urinalysis
o Both statements are correct
o Rinsing or washing of glassware
o None of the statements are correct
34. A flameless atomic absorption
spectrophotomter dehydrates and atomizes a
sample using
o An electon gun
o A thermospray platform
o A thermoelectric semiconductor
o A graphite capillary furnace
35. Chromatography is a physical method that is
used to separate and analyze
o Viscous mixtures
o Complex mixtures
o Metals
o Simple mixtures
36. The term reverse phase is used in HPLC to
indicate that the mobile phase is
o More polar than the stationary phase
o A stronger solvent than the stationary
phase
o Liquid and the stationary phase is
solid
o Organic and the stationary phase is
aqueous
37. Which component is required in a
spectrophotometer in order to produce a
spectral absorbance curve?
o Multiple monochromators
o Laser light source
o Photodiode array
o A reference optical beam
38. The term isocratic is used in HPLC to mean
the
o Flow rate of the mobile phase is regulated
o Mobile phase consists of constant solvent
composition
o Stationary phase is equilibrated with the
mobile phase
o Mobile phase is at constant temperature
39. Which type of filter is best for measuring stray
light?
o Written
o Neutral density
o Didymium
o Sharp cutoff
40. Which of the ff. is most commonly used detector
for clinical gas-liquid chromatography?
o Flame ionization
o Both items are correct
o Thermal conductance
o Neither of these items are correct
41. Which condition is a common cause of stray light?
o Dispersion from second-order spectra
o Improper wavelength calibration
o Unstable source lamp voltage
o Misaligned source lamp
42. In thin-layer chromatography, the distance the
solute migrates divided by the distance the
solvent migrates is the
o pK
o tR
o Rf
o Kd
43. Which photodetector is most sensitive to low
levels of light?
o Photomultiplier tube
o Photodiode
o Barrier layer cell
o Diode array
44. Why is vacuum system necessary in the mass filter
of a mass spectrophotometer?
o It prevents collision between
molecules/ions
o Ionization doesn’t occur at atmospheric
pressure
o It remove electrons form the ion source
o It prevents contamination
45. Which type of monochromator produces the
purest monochromatic light in the UV range?
o Interference filter and a variable exit slit
o A prism and a variable exit slit
o A sharp cutoff filter and a variable exit slit
o A diffraction grading and a fixed exit slit
46. In gas chromatography, the elution order of o The gas phase
volatiles is usually based upon the o The stationary phase
o Carbon content o The liquid phase
o Molecular size 54. Which of the ff. is not a problem inherent in
o Boiling point turbidimetry?
o Polarity o Need to maintain a constant and specific
47. Which of the following components is not temperature
needed in a chemiluminiscent immunoassay o Variation in particle size of samples
analyzer? o Variation in particle size of standards
o Photodetector o Rate of aggregation or settling of particles
o Source lamp 55. Fluorometers are designed so that the path of the
o Monochromator exciting light is at a right angle to the path of the
o Wash station emitted light. What is the purpose of this design?
48. Most commonly used solvents for the mobile o Prevent loss of the excitation light
phase in most HPLC is o Percent excitation light from reaching
o Acteonitrite the detector
o Hexane o Prevent loss of emitted light
o Chloroform o Focus emitted and excitation light upon
o Ethyl acetate the detector
49. Setting the spectrophotometer to zero is 56. Which of the ff represents a primary and
done using advantage of performing fluorometric over
o Water blank absorption spectroscopic methods of analysis?
o Reagent blank Ease of performing assays
o Sample blank Increased specificity and decreased sensitivity
o Any of the options is correct Increased specificity and increased sensitivity
50. Checked with standard absorbing solution or Purity of reagents used not as critical
filters 57. Which of the ff. may be associated with
o Stray light bioluminescence?
o Linearity o Electron excitation caused by radiant
o Wavelength energy
o All options are correct o Employs a radioactive label
51. What electrochemical technique measures o Less sensitive than direct fluorescent
the current produced through oxidation or assays
reduction between the indicator and o Light emission produced due to
reference electrodes held at a constant enzymatic oxidation of a substrate
electrical potential (voltage) in an 58. Which of the ff. best describes
electrochemical cell? chemiluminiscence?
o Amperometry o Electron excitation caused by radiant
o Coulometry energy
o Potentiometry o Enzymatic oxidation of a substrate
o Conductometry produces light emission
o Employs a radioactive label
o Chemical energy excites electrons that
emit light upon return to ground state.
52. What physical property is used to separate
59. Nephelometry is based on the measurement of
complex mixtures in chromatography?
light that is
o Density
o Absorbed by particles in suspension
o Hydrogen ion concentration
o Produced by excitation of ground state
o Vapor pressure
atoms
o Differential solubility
o Produced by fluorescence
53. The solvent is what part of a thin layer of
o Scattered by particles in suspension
chromatography system
o The mobile phase
60. In potentiometry, which of the ff. is
considered the standard electrode?
o Hydrogen electrode
o Calcium electrode
o Copper electrode
o Potassium electrode

Nephelometry
• The measurement of light scattered by a particulate
solution (big particles)
• Generally, scattered light is measured at an angle to
the incident light when small particles are involved;
for large molecules, forward light scatter can be
measured
• The amount of scatter is directly proportional to
the number and size of particles present in the
solution
• The sensitivity of nephelometry depends on the
absence of background scatter from scratched
cuvets and particulate matter in reagents

Turbidimetry
• Measures light blocked as a decrease in the light
transmitted through the solution, dependent on
particle size and concentration
• Uses a spectrophotometer for measurement, and
is limited by photometric accuracy and sensitivity of
the instrument
• The problems inherent in turbidimetry
- Variation in particle size of samples
- Variation in particle size of standards
- Rate of aggregation or settling of particles
Molecular Emission Spectroscopy
• Type of luminescence where excitation requires
absorption of radiant energy
- Fluorescence – emission of light by a substance
that has absorbed light or other electromagnetic
radiation
➢ Emitted light has a longer wavelength, and
therefore, lower energy than the absorbed
radiation
➢ Fluorometry – measurement of the
emitted fluorescence light
➢ The excitation light is absorbed by the
atoms of the analyte in solution, which
causes the electrons to move to higher
energy orbitals
➢ Upon return to ground state, light is
emitted from the fluorescing analyte and
that light passes through a secondary filter.
➢ The secondary filter and the detector are
placed at a right angle to the light source to
prevent incident light from being
measured by the detector
➢ Fluorometers use filters,
spectrofluorometers use prisms or
diffraction gratings as monochromators
➢ Advantage – fluorometry is about 1000x
more sensitive than absorption techniques
and has increased specificity because
optimal wavelengths are chosen both for
absorption (excitation) and for monitoring
emitted fluorescence
➢ Limitations – changes from the established
protocol that affect pH, temperature, and
solvent quality; self-absorption; quenching
➢ Fluorometers are designed so that the
path of exciting light is at a right angle
to the path of the emitted light and
this design prevents excitation light
from reaching the detector
➢ Requires primary and secondary
monochromator
➢ Primary advantage of performing
fluorometric over absorption
spectrometric methods of analysis is
increased specificity and increased
sensitivity
- Phosphorescence – emission of light
produced by certain substances after
theyabsorb energy
➢ It is similar to fluorescence except
that the time delay is longer (greater
than 10-4 sec) between absorption of
radiant energy as photons of light
• Types of luminescence where excitation does not
require absorption of radiant energy
- Chemoluminescence – chemical energy of a
reaction produces excited atoms and upon
electron return to ground state, photons of
light are emitted.
➢ Advantage – has excellent sensitivity and
dynamic range
➢ Ruthenium is the substance used to
generate light signal
➢ Source lamp is not needed in
chemiluminescent immunoassay, has
monochromator, photodetector and wash
solution
- Bioluminescence – enzyme-catalyzed chemical
reaction produces light emission
➢ This may occur in the presence of the
enzyme luciferase because of oxidation of
the substrate luciferin
➢ Luminometer is a generic term for the type
of instrument that is used to measure
chemiluminesce andbioluminescence
Chromatography
• Technique where solutes in a sample are separated
for identification based on physical differences
that allow their differential distribution between a
mobile phase and a stationary phase
• Mobile phase may be an inert gas or liquid
• Stationary phase may be silica gel bound to the
surface of a glass plate or plastic sheet; may be
silica or a polymer that is coated or bonded within a
column
Thin Layer Chromatography
- Type of planar chromatography
- The stationary phase may be silica gel that is
coated onto a solid surface such as a glass plate
or plastic sheet
- The mobile phase is a solvent, where solvent
polarity should be just enough to achieve clear
separation of the solutes in the sample
- Used clinically for urine drug screening
- The mobile phase moves through the
stationary phase by absorption and capillary
action
- The solute components move at different rates
because of solubility in the mobile phase that
retard solute movement
- These two phases work together to provide solute
resolution and separation
 ➢ Solute will stay with the solvent front if
solvent is too polar for the solute
➢ Solute will remain at origin if solvent is
insufficiently polar
- Basic steps in performing TLC include
➢ Sample extraction using a liquid-liquid or
column technique
➢ Concentration of the extracted sample
➢ Sample application by spotting onto the
silica gel plate
➢ Development of the solute in the sample
using stationary and mobile phases
➢ Solute detection using chromogenic
sprays, UV light, fluorescence, and heat
➢ Interpretation of chromatographic results
utilizing Rf values of solutes in comparison
to aqueous standards
- In TLC, Rf is the distance the solute migrates
divided by the distance the solvent migrates
- Rf values are affected by chamber saturation,
temperature, humidity, and composition of the
solvent
Gas-Liquid Chromatography
• Components include
- A carrier gas with a flow-control device to
regulate the gas flow
- A heated injected injector
- Chromatographic column to separate the solute
- Heated column oven
- Detector
- Computer to process the data and control the
operation of the system
• Gas-liquid chromatography is a technique used to
separate volatile solutes
High-Performance Liquid Chromatography (HPLC)
• HPLC components include
- Sample-introduction system
- Solvent reservoirs (solvent-delivery) system
- One or more pumps to propel the solvent(s)
- Injector
- Chromatographic column
- Detector
- Computer to process data and control the
operation of the system
• HPLC is a type of liquid chromatography
where the mobile phase is a liquid that is
passed over the stationary phase of the
column
- The separation of solutes in a sample is
governed by the selective distribution of
the solutes between the mobile and the
stationary phase
- Solvents commonly used for the mobile phase
include acetonitrite, methanol, ethanol,
isopropanol and water
➢ Isocratic elution – strength of solvent
remains constant during separation
➢ Gradient elution – strength of solvent
continually increases (%/min) during
separation
- Stationary phase is an organic material
covalently bonded to silica that may be polar or
nonpolar in composition
➢ Normal-phase liquid chromatography –
polar stationary phase and nonpolar
mobile phase
➢ Reversed-phase liquid chromatography –
nonpolar stationary and polar mobile phase
- The solvent-delivery system utilizes a solvent
reservoir from which the pump can punch the
mobile phase through the column
➢ The sample is introduced through a loop
injector
➢ A pre-column and guard column function to
maintain the integrity of the column and are
positioned prior to the sample reaching the
main column
➢ The column, which functions as the stationary
phase, generally operates at room temperature
➢ The effluent from the column passes to a
detector system
➢ The solutes are introduced to the detector in the
order that each was eluted
- The detector produces a signal for identification and
quantification of the solutes
➢ Commonly used detectors include
spectrophotometer, photodiode array,
fluorometer, electrochemical (glassy carbon
electrode)
➢ It separates solutes in a sample based on the sign
and magnitude of the ionic charge
Mass Spectrometry
• A mass spectrometer is an instrument that uses the
principle of charged particles moving through a
magnetic or electric field with ions being separated
from other charged particles according to their mass- to-
charge ratios (parameter used to identify a compound)
- In this system, electrons bombard a sample, ionizing
the compound into fragment ions, which are separated
by their mass-to-charge ratios
- The mass spectrum produced is unique for a
compound (identification), and the number of ions
produced relates proportionally to concentration
 (quantification)
• Mass spectrometry is a high-quality technique for
identifying drugs or drug metabolites, amino acid
composition of proteins, and steroids
- In addition, mass spectrometry has applications in
the field of proteomics
- The eluate gas from a gas chromatograph may be
introduced into a mass spectrometer that
functions as the detector system, or the liquid
eluate may be introduced from a high-liquid
chromatograph
• Instrumentation – mass spectrometer
components include
- Ion source – samples enter the ion source and are
bombarded by the ionization beam
➢ When the sample is in gas form and
introduced from a gas chromatograph, the
ion source may be electron or chemical
ionization
➢ Other types, such as electrospray
ionization and sonic spray ionization, may
be used when a high-performance liquid
chromatograph is used in conjunction with
spectrometer
- Vacuum system – prevents the collision of ions
with other molecules when electronic or magnetic
separation is occurring
- Analyzer – beam-type and trapping-type
➢ Beam-type is a destructive process,
where ions pass through the analyzer
one time and then strike the detector
➢ Quadropole is a beam-type analyzer,
where mass-to-charge ratios are
scanned during a prescribed time
period to form a mass spectrum
- Detector usually detects ions using
electron multipliers, such as discrete
dynode and continuous dynode electron
multipliers
- Computer and software convert the
detector’s signal to a digital form
➢ Sample identification is achieved
because each compound produces a
unique spectrum, which is analyzed by
a database for matching to a
computerized reference library
• To further improve selectivity and sensitivity,
a system known as tandem mass
spectrometers can be employed, where a gas
chromatograph is connected to two mass
spectrometers (GC MS/MS) or (HPLC/MS/MS)
- In these systems, ions of a specific mass-to-
charge ratio are allowed to continue to the
second mass spectrometer where additional
fragmentation occurs and final analysis is done
- The sample injected into the injector component of the
instrument where the sample is vaporized because the
injector is maintained approximately 50°C higher than
the column temperature
- An inert carrier gas (mobile phase) carries the
vaporized sample into the column. Carrier gases
commonly used include hydrogen, helium, nitrogen,
and argon.
➢ The carrier gas flow rate is critical to maintaining
column efficiency and reproducibility of elution
times.
➢ The elution order of volatiles is usually based
upon the boiling point
- The types of columns (stationary phase) used are
designated as packed or capillary
➢ When the volatile solutes carried by the gas over
the stationary phase of the column are eluted,
the column effluent is introduced to the
detector
➢ The solutes are introduced to the detector in the
order that each was eluted
➢The detector produces a signal for
identification and quantification of the
solutes Commonly used detectors include
flame ionization, thermal conductivity,
electron capture and mass spectrometer
- Separator of solutes is a function of the relative
differences between the vapor pressure of the
solutes and the interactions of the solutes with the
stationary column
➢ The more volatile a solute, the faster it will
elute from the column; the less interaction
of the solute with the column, the faster it
will elute
- Identification of a solute is based on its
retention time, and quantification is based on
peak size, where the amount of solute present is
proportional to the size of the peak (area of
height of the sample peak, is compared to the
known standard)
➢ In addition, GLC…
o Separation depends on the sample and
solubility in the liquid layer of the
stationary phase
o Stationary phase is a liquid layer
absorbed on the column packing
Reflectance Spectrophotometry
• Associated with unabsorbed, reflected light
detected by the photodetector as it relates to the dry
 reagent slide technique
• Measures the concentration of glucose in the
blood by using dry film technology
• Associated with plane-polarized light for
sample excitation
Fluorescence Polarization
• When the sample (fluorophore) is excited, it
emits polarized light among the same plane as
the incident light if the fluorophore does not
rotate in solution (i.e., it is attached or bound to
a large molecule)
• In contrast, a small molecule emits
depolarized light because it will rotate out of
the plane of polarization during its excitation
lifetime
• This technique is widely used for the
detection of therapeutic and abuseddrugs
• In the procedure, the sample analyte is
allowed to compete with a fluorophore-
labeled analyte for a limites antibody to
analyte
• The lower the concentration of the sample
analyte, the higher the macromolecular
antibody-analyte- fluorophore formed and
lower the depolarization of the radiant light
Electrochemistry
• When chemical energy is converted into an
electrical current (flow of electrons) in a
galvanic cell, the term electrochemistry is used
• Electrochemical reactions are characterized by
a loss of electrons (oxidation) at the
positive pole (anode) and a simultaneous
gain of electrons (reduction) at the negative
pole (cathode)
• The galvanic cell is made up of two parts called half-
cells, each containing a metal in a solution of one of its
salts
• These methods involve the measurement of
electrical signals associated with chemical systems
that are within an electrochemical cell
• In the clinical lab, electroanalytical methods are used
to measure ions, drugs, hormones, metals and gases
• Methods are available for the rapid analysis of
analytes present in relatively high concentrationsin
blood and urine, such as blood electrolytes (Na+, K+,
Cl-
, HCO3-), and other analytes present in very low
concentrations, such as heavy metals and drug
metabolites
• Many types of electrochemical analyses are used
including potentiometry, amperometry,
coulometry, and polarometry
• The two basic electrochemical cells involved in these
analyses are galvanic and electrolytic cells
• Polarography – analytical procedure measuring
concentration of a substance based on the amount of
current flow resulting from ion transfer when
substance go into chemical reaction; employs an
electrochemical cell
- Gradually increasing the voltage applied between two
electrodes of the cell in contact with a solution
containing the analyte
- Current measured – voltage change versus plotted to
produce a polarogram
- Voltage at which sharp rise in current occurs
characteristic of the electrochemical reaction involved
- Amount of increase in current (i.e., wave height)
proportional to the concentration of analyte
• Anodic voltammetry is based on polarography
- Negativepotentialappliedtooneoftheelectrodes
- Trace metal ions in the solution reduced and plated
onto anodic electrode;pre-concentrating step
- Plated electrode used as anode in polarographic cell;
metal stripped off anode
- Current flow during stripping provides polarogram
that identifies and quantifies the analyte being
measured (trace metals)
- Used to assay heavy metals such as lead in blood
• Potentiometry – technique used to determine the
concentration of a substance in solution employing an
electrochemical cell that consists of two half-cells,
where the potential difference between an indicator
electrode and a reference electrode is measured
- Half-cell, also called an electrode, composed of single
metallic conductor surrounded by solution of
electrolyte
- In an electrolytic cell, the half cell where reduction takes
place is a cathode
- Two different half-cells connected to make complete
circuit; current flows because of potential difference
between two electrodes
- Salt bridge connection between two metallic
conductors and between two electrolyte
solutions
- Comparison made between the voltage of one
half-cell compared to another half-cell
- Half-cell potentials compared to potential
generated by standard electrode
- Universally accepted standard half-cell is the
standard hydrogen electrode, arbitrally
assigned a potential E0 of 0.000 volt
- Desirable to use one half-cell (reference electrode)
with known and constant potential, not
sensitive to composition of material to be
 analyzed
- Calomel electrode – type of reference
electrode, consisting of mercury covered
by a layer of mercurous chloride in contact
with saturated solution of potassium
chloride
- Other half-cell (indicator electrode)
selected on basis of change in its potential
with change in concentration of analyte to
be measured
- Silver-silver chloride (Ag/AgCl) electrode;
common type of reference electrode
• A pH/blood gas analyzer employs a pH-
sensitive glass electrode for measuring blood
pH, and employs PCO2 and PO2 electrodes for
measuring gases in blood
- For measuring pH, the pH electrode is a
functioning glass electrode that is
dependent on properties of pH-sensitive
glass
- When a pH-sensitive glass electrode is not
actively in use, it should be kept in a
medium recommended by the
manufacturer
- To calibrate the pH electrode, it is
necessary that calibrating gases, two
buffer solutions of known pH be used
➢ Glass electrode made by sealing thin
piece of pH-sensitive glass at the end
of glass tubing and filling tube with
solution of HCl saturated with AgCl
➢ Silver wire immersed in tube’s
solution with one end extending
outside the tube for external
connection, silver-silver chloride
reference electrode sealed within
tube with pH-sensitive glass tip
➢ pH sensitive glass must be saturated
with water. Surface of the glass
develops a hydrated lattice, allowing
exchange of alkaline metal ions in the
lattice for hydrogen ions in the test
solution
o A potential is created between the
inside and the outside of the
electrode, and that potential is
measured
➢ Glass electrode calibrated by
comparison with two primary
standard buffers of known pH
➢ Because pH readings are temperature
sensitive, the calibration must be
carried out at a constant temperature
of 37°C
- In a pH/blood gas analyzer, the PCO2 electrode
for measuring the partial pressure of carbon
dioxide (PCO2) in blood is actually a pH electrode
immersed in a bicarbonate solution
➢ The bicarbonate solution is separated from
the sample by a membrane that is
permeable in gaseous CO2 but not to
ionized substance such as H+ ions
➢ When CO2 from the sample diffuses across the
membrane,itdissolves,formingcarbonicacid and
thus lowering the pH
o The measurement of CO2 in blood b means
of PCO2 electrode is dependent on the
change in pH because of increased carbonic
acid in the electrolyte surrounding the
electrodes
o The pH is inversely proportional to the log of
the PCO2. Hence, the scale of meter can be
calibrates directly in terms of PCO2
- The ion-exchange electrode is a type of
potentiometric, ion-selective electrode
➢ The ion-selective electrode universally used is the
pH electrode
o Consists of liquid ion-exchange membrane
made of inert solvent and ion- selective
neutral carrier material
➢ Colloidon membrane may be used to separate
membrane solution from sample solution
➢ K+ analysis – antibiotic valinomycin, because of
its ability to bind K+ used as a natural carrier for
K+ selective membrane
➢ NH4+ analysis – antibiotics nonactin and
monactin used in combination as neutral carrier
for NH4+-selective membrane
- Sodium analysis – ion-selective electrodes based
on principle of potentiometry
➢ Unlike glass membrane, electrodes with
selective capability
➢ Constructed from glass that consists of silicon
dioxide, sodium oxide and aluminum oxide
➢ Ion-selective electrode analysis of sodium
o Uses a glass membrane
o Error occur from protein build-up on the
membrane
o Principle is based on potentiometry
o Membrane not coated with valinomycin
• Amperometry – electrochemical technique that
measures the amount of current produced through the
oxidation or reduction of the substance to be measured
at an electrode held at fixed potential
- In a pH/blood analyzer, the electrode for measuring
the PO2 in the blood is an electrochemical cell
consisting of a platinum cathode and a Ag/AgCl anode
 connected to an external voltage source
- The cathode and anode are immersed in the
buffer. A polyropylene membrane selectively
permeable to gases separates the electrodes and
buffer from the sample
- When there is no oxygen diffusing into the buffer,
there is practically no current flowing between the
cathodeand the anode because they are polarized
- When oxygen diffuses into the buffer from a
sample, it is reduced from the platinum cathode
- The electrons necessary for this reduction are
produced at the anode. Hence, aa current flows;
the current is directly proportional to the
PO2 in the sample
• Coulometry
- A chloride coulometry employs a
soulometric system based on Faraday’s law,
which states that in an electrochemical
system, the number of equivalent weight of
a reactant oxidized or reduced is directly
proportional to the quantity of electricity
used in the reaction
- The quantity of electricity is measured in
coulombs. Coulomb is the unit of electrical
quality. I coulomb of electricity flowing per
minute constitutes a current of I ampere
- If the current is constant, the number of
equivalent weights of reactant oxidized or
reduced depends only on the duration of
the current
- In the chloride coulometer, the
electrochemical reaction is the generation
of Ag ions by the passage of a direct current
across a pair of silver electrodes immersed in
a conducting solution containing the sample
to be assayed for chloride
➢ As the Ag+ are generated, they are
immediately removed from solution
by combining with chloride to form
insoluble AgCl
➢ When all the chloride is precipitated;
further generation of Ag+ causes an
increase in conductivity of the solution
- The endpoint of the titration is indicated by
the increase in conductivity of the solution
Coulometric chloridometers and Anodic stripping
voltammetry
• Chloride ISEs have largely replaced
coulometric titrations for the determination
of chloride in body fluids
• Anodic stripping voltammetry was widely
used for the analysis of lead and is best
measured by electrothermal (graphic furnace),
atomic absorption spectroscopy or, preferably ICP-
M
Electrophoresis
• Used clinically to separate and identify proteins,
including serum, urine and CSF proteins,
lipoproteins, isoenzymes and so on
• Movement of charged molecules in a medium when
an electric field is applied
• Zone electrophoresis is defined as the movement
of charged molecules in a porous supporting
medium where the molecules separate as distinct
zones
• Support medium provides a matrix that allows
molecules to separate (e.g., agarose gel, starchgel,
polyacrylamide gel and cellulose acetate
membranes)
• Movement of charged particles through a medium
depends on the nature of the particle, including net
charge, size and shape, the character of the buffer
and supporting medium, temperature and the
intensity of the electric field
- Nature of the charged particle – proteins are
amphoteric and may be charged positively or
negatively depending on the pH of the buffer solution
- The pH at which negative and positive charges are equal
on a protein is a protein’s isoelectric point
• Buffer solutions of pH 8.6 are generally used for serum
protein electrophoresis. Using agarose gel or cellulose
acetate at this alkaline pH, serum proteins take on a net
negative charge and will migrate toward the anode (+)
- Albumin migrates the fastest toward the anode and
the gamma globulins remain closer to the cathode (-)
• Visualizing the separated analyte – following
electrophoresis, treat the support medium with
colorimetric stains or fluorescent chemicals
- Amido black B, Ponceau S and Coomassie brilliant
blue stains are used for visualizing serum proteins
- Silver nitrate is used for CSF proteins, fat red 7B and oil
red O are used for lipoproteins and nitrotetrazolium
blue is used for lactate dehydrogenase
• Detection and quantification of the separated protein is
accomplished using densitometer
• Commonly encountered problems in electrophoresis
- Holes in staining pattern – analyte present in too high
concentrations
- Very slow migration – voltage too low
- Sample precipitates in support – pH too high or too
low; excessive heat production
• Isoelectric focusing is a type of zone electrophoresis in
which protein separation is based on the isoelectric
point (pI) of the proteins
 -This method utilizes polyacrilamide or agarose gel
containing a pH gradient formed by ampholytes in
the medium
- When exposed to an electric field, the ampholytes
migrate based on their pI to their respective
positions in the gradient. In turn, the serum
proteins will migrate in the gel to the position
where the gel’s pH equals the pI of the respective
protein
• Capillary electrophoresis is based on electrosonic
flow (EOF) – when an electric field is applied, the flow
of liquid is in the direction of the cathode. Thus, EOF
regulates the speed at which solutes move through
the capillary
Automation Parameters/Terminology
• Centrifugal analysis – centrifugal force moves
samples and reagents into cuvet areas for
aimultaneous analsis
• Discrete analysis – each sample reaction is
compartmentalized
This may relate to an analyzer designed to
assay only one analyte (e.g. glucose) or an
analyzer capable of forming multiple
tests where the sample and reagents are
in a separate cuvet/reaction vessel for
each test
• Random access – able to perform individual
tests or panels, and allows for stat samples to
be added to run ahead of other specimens
• Batch analysis – samples processed as a group
• Stand alone – instrument from a single disciple
with automated capability
• Automated stand alone – instrument from a
single discipline with internal automated
capability (e.g. auto-repeat and auto-dilute)
• Modular workcell – at least two instruments
from a single discipline with additional
automated capability
• Multiple platform – instrument able to
perform tests from at least two disciplines
• Integrated modular system – at least two
analytical modules supported by one sample
and reagent processing and delivery system
• Pneumatic tube system – transports
specimens quickly from one location to
another
• Throughput – maximum number of tests
generated per hour
• Turnaround – amount of time to generate one result
• Bar coding – mechanism for patient/sample
identifications; used for reagent identification
by an instrument
• Dead volume – amount of serum that cannot
be aspirated
• Carry-over – the contamination of a sample by a
previously aspirated sample
• Reflex testing – use of preliminary test results to
determine if additional tests should be ordered or
cancelled on a particular specimen; performed
manually or automated
• Total laboratory automation – automated systems
exist for laboratories where samples are received,
centrifuged, distributed to particular instruments
using a conveyor system, and loaded into the
analyzer without operator assistance
- This kind of automation is seen in large medical
center laboratories where the volume of testing
is high
Principles of Automation
• Automated instruments use robotics and fluidics
to replicate manual tasks
• Specimen handling – some instruments have level-
sending probes that detect the amount of serum or
plasma in the tube
- Some systems have a reading device that
allows bar-coded sample tubes to be loaded
onto the instrument
- Although not as common, other instruments
require the operator to manually enter the
position of the patient sample
• Reagents
- Dry reagents can be packaged a lyophilized
powder or tablet form that must be
reconstituted with a buffer or reagent-grade
water
- Reconstituting of reagents may need to be done
manually and then the reagents placed on an analyzer
for use, or reconstituting the reagents as employed by
Dimension® analyzer
- Dry reagents can be spread over a support material
and assemble into a single-use slide. This technique is
employed by Vitros® analyzer
- Liquid reagents are pipette by the instrument and
mixed with the sample
• Testing Phase
- Mixing sample and reagents occurs in a vessel called a
cuvet. Come instruments have permanent,
nondisposable cuvets made of quartz glass. Other
cuvets are made of plastic and are disposable
- Reaction temperatures and times vary for each
analyte. The most common reaction temperatures are
37°C and 30°C
- Kinetic assays – determination of sample
concentration is based on change in absorbance over
 time instrument. The program must use appropriate
- Endpoint/colometric assays – incubated for a standards and controls, statistical analyses, and a
specific time, absorbance determined, absorbance proficiency testing system
related to calibrators for calculation of sample - This information must be documented
concentration
- A spectrometer is built within the system to read
absorbances for kinetic and colometric assays
➢ These systems may use a diffraction
grating or aa series of high quality fibers.
Some automated analyzers incorporate
fluorometry or nephelometry
• Data Management
- The computer module of most automated
instruments has a data management system that
allows analysis of quality control (QC) materials
and assessment of patient values (e.g. delta check)
before releasing patient results
- Instruments/laboratory information systems
(LISs) also archive patient results and QC values.
These achieved results are stored by the
laboratory for various lengths of time

Point-of-Care Testing (POCT)

• Definition – performing diagnostic tests outside
the main laboratory and at or near patient care areas
• Applications – POCT is designed to provide
immediate laboratory test results for immediate
patient assessment and determination of
appropriate treatment
- POCT may be used in neonatal intensive care,
coronary care, intensive care or the emergency
department
• Operators
- Only waived laboratory tests can be performed
using POC instruments
- Clinical laboratory technicians and clinical
laboratory scientists must operate instruments
that perform complex or high-complexity
laboratory tests
• POC instrument evaluation
- All POC instruments must be evaluated in
accordance with the CLIA ‘88
- The values obtained from POC instruments
must correlate with values obtained from
larger laboratory instruments
- Linearity testing, calculation of control
ranges, correlations of sample data and
reference ranges must be done for each
instrument
• Training – all POC instrument operators must
be trained, and training must be documented
• Quality control – all effective quality control
systems must be set up for each POC
 • Qualified personnel
- Only properly certified personnel can perform
nonwaived assays
- They must be able to perform QC activities,
maintain instruments, and keep accurate and
systematic records of reagents and control
specimens, equipment maintenance, and
patient and analytical data
- They must have in-training service classes and
opportunities to attend a continuing education
program
Quality Assurance and Quality Nonanalytical Factors
- Periodic opportunities for personal upgrading
Control of technical skills and for obtaining new
Quality Assurance relevant information should be made available
to all persons working in the lab.
- A way of ensuring that - Personnel performance should be monitored
with periodic evaluations and reports.
quality requirements will • Established laboratory policies
- an updated manual contains a comprehensive
be fulfilled – the product listing of approved policies, acceptable
will be “fit for purpose” practices and precautions, including standard
blood and body fluid precautions
and made “right the first - should be included in a lab reference manual that is
available to all hospital personnel.
time”. - Each lab must have an up-to-date safety manual.
Contains comprehensive listing of approved
Quality Control
policies, acceptable practices, and standard
precautions (blood and body fluids).
- A process of fulfilling - OSHA and CDC
• Laboratory procedure manual
quality requirements by - this manual must be reviewed regularly by the
thoroughly reviewing the supervisor and updated, as needed.
- Should follow a specific pattern in how the
quality of all factors procedures are organized (CLSI).
- Minimal components:
involved in production. Title of the assay
Principle of the procedure & statement of clinical
Quality Assessment
application
• Currently, the majority of laboratory errors are
Protocol for specimen collection and storage
related to pre-analytical or post-analytical
Nonanalytical Factors
phases of testing. Specimen-related errors • QC information
continue to be a major problem, leading to • Reagents, supplies and equipment
unnecessary costs to hospitals • Procedural protocol
• To reduce and potentially eliminate laboratory • “Normal” reference ranges
errors, a quality assessment program is • Technical sources of error
• Limitations of the procedure
mandated
• Proper procedures for specimen collection and storage
• Two major components of quality assessment
program
- Non-analytical factors
- Analysis of quantitative data (quality control)

Non-analytical Factors
 regulations.
- Test request procedures, patient identification,
specimen procurement and labeling; specimen
transportation and processing procedures; lab
personnel performance; lab instrumentation, reagents
• Test questioning
and analytical test procedures; turn around times, and
- Lab test can be requested by a primary care
the accuracy of the final results.
provider or patient.
• Proficiency Testing (PT)
- the request, either hard copy or electronic,
- Identical samples are sent to a group of laboratories
must include all information about the test
participating in the PT program; each lab analyzes the
request, patient’s data accompanied by the
specimen, reports the results to the agency, and is
specimen to be tested
evaluated and graded on those results in comparison
- The information on the accompanying
to the results from other laboratories.
specimen container must match exactly the
• Interpretation of the Results of the Proficiency Testing
patient identification on the test request – (PT):
included in a database or handbook. - Each analyte has a define performance criteria
• Patient Identification, Specimen Procurement and
(example, +/-3SD of peer mean), where laboratories
Labeling using the same method are evaluated by comparing
- current information about obtaining appropriate
them with the group.
specimens, special collection requirements of - In external QC , difference of greater than 2SD in the
various types of tests, ordering tests correctly results indicates that a laboratory is not in agreement
and transporting and processing of specimens with the rest of the laboratories included in the
appropriately should be included in the database. program.
• Specimen Transportation and Processing
- If in case a clinical laboratory failed to identify or
- Some assays require special handling conditions,
resolve an error or discrepancy in the test process, the
such as placing the specimen on ice immediately facility is at risk of continuous operation and may be
after collection. recommended for closure.
• Accuracy in Reporting Results and Documentation:
- Specimens should be tested within 2 hours after - Many laboratories have established critical values or
collection to produce accurate results. the Delta check system to monitor individual patient
results.
- The documentation of specimen arrival times in - Highly abnormal individual test values and significant
the lab as well other specific test request data is differences from previous results in the Delta check
an important aspect of the quality assessment system alert the technologist to a potential problem.
process. - The ongoing process of making certain that the correct
• Preventive Maintenance of Equipment lab result is reported for the right patient in a timely
- Monitoring of the temperatures of the heat manner and at the correct cost is known as continuous
blocks and refrigerators is important in the quality improvement (CQI).
quality of test performance.
- Microscopes, centrifuges and other equipment
need regularly to be cleaned and checked for
accuracy.
- Mandatory recalibration of instrument systems.
• Appropriate Testing Methods
- Each lab must have an assessment routine for all
lab procedures, performed on a daily, weekly or
monthly basis to detect problems such as trends
and shifts in the established mean values.
- When such problems are indicated, they must be
corrected as soon as possible.
• Quality Assessment Procedures
- The documentation of an ongoing quality Quality Control (QC)
assessment program is mandated by CLIA
 • A system of ensuring accuracy & precision in the
lab by including quality control reagents in every
series of measurements.
• A process of ensuring that analytical results are
correct by testing known samples that resemble
patient samples.
• It involves the process of monitoring the
characteristics of the analytical processes and
detects analytical errors during testing and
ultimately prevent the reporting of inaccurate
patient test results.
• It is one component of the quality assurance
system and is part of the performance
monitoring test that occurs after a test has
been established.
• Consists of procedures used to detect error that
result from test system failure, adverse
environmental conditions, and variance in operator
performance, as
well as procedures to monitor the accuracy and
precision of test performance over time
• Accrediting agencies require monitoring and
documentation of quality assessment records
• QC activities include monitoring the performance
of laboratory instruments, reagents, other testing
products and equipment
• A written record of QC activities for each procedure
or function should include details of deviation from
the usual results, problems, or failures in functioning
or in the analytical procedure and any corrective
action taken in response to these problems
• Documentation of QC includes preventive
maintenance records, temperature charts, and QC
charts for specific assays
• All products and reagents used in the analytical
procedures must be carefully checked before
actual use in testing patient samples
• Use of QC specimens, proficiency testing and
standards depends on the specific requirements of
the accrediting agency
Parameters
• Sensitivity – ability of an analytical method to
measure the smallest concentration of the analyte of
interest.
• Specificity – ability of an analytical method to
measure only the analyte of interest
 - varies from sample to sample.
- basis for varying differences between repeated
measurements – variations in technique.
- due to instrument, operator, and environmental
conditions (variations in technique) such as pipetting
error, mislabeling of samples, temperature
fluctuation, and improper mixing of sample and
reagent.

2.Systematic Error
- error that influences observations consistently in one
direction (constant difference).
- detected as either positive or negative bias – often related
to calibration problems, deterioration of reagents and
control materials, improperly made standard solutions,
contaminated solutions, unstable and inadequate reagent
• Practicability - is the degree by which a method
blanks, leaky ion selective electrodes, failing instrumentation
is easily repeated.
and poorly written procedures.
• Reliability – the ability of an analytical method
to maintain accuracy and precision over an
extended period of time during which a. Constant Error
equipment , reagents, and personnel may - refers to the difference between the target value and the
change. assayed value.
Kinds of Quality Control - independent of sample concentration.
Intralab Quality Control (internal QC) - exists when there is continual difference between the
• involves the analysis of control samples together comparative method and the test method regardless of the
with the patient specimens. concentration.
• detects changes in performance between the
present operation and the “stable” operation.
• important for the daily monitoring of accuracy
and precision of analytical methods.
• detects both random and systematic errors in a
daily basis.
Interlab Quality Control (external QC)
• involves proficiency testing programs that
periodically provide samples of unknown
concentrations to participating clinical
laboratories.
• important in maintaining long-term accuracy of
the analytical methods.
• used to determine state-of-the-art
interlaboratory performance.
• the College of American Pathologist (CAP)
proficiency program is the gold standard for b. Proportional/Slope/Percent Error
clinical lab external QC testing. - results in greater deviation from the target value due to
Variations higher sample concentration.
Errors encountered in the collection, preparation and - exists when the difference between the test method and
measurement of samples, including transcription and the comparative method values is proportional to the
analyte concentration.
releasing of laboratory results.

Types of Variations 3. Clerical Error

1.Random Error - the highest frequency of clerical errors occurs with thee
- present in all measurements, it is due to use of handwritten labels and request forms.
chance.

Pre-analytical Errors – occur prior to the testing
process
- incorrect patient identification
- improper patient preparation
- incorrect specimen collection
- mislabeled specimen
- incorrect order of draw
- incorrect use of tubes for blood collection
- incorrect anticoagulant to blood ratio
- improper mixing of blood and anticoagulant
- incorrect specimen preservation
- mishandled specimen (transport and storage)
- incorrectly interpreted/ordered lab test
- incomplete centrifugation
- incorrect data log-in
Analytical Errors – occur within the laboratory during
the testing process.
- incorrect sample and reagent volume
- incorrect incubation of solution
- equipment/instrument malfunction
- improper calibration of equipment/calibration error
Post-analytical Errors – occur after a test result is
generated.
- unavailable or delayed lab results
- long turnaround time
- wrong transcription of the patient’s data and lab
results
- missing lab results
- lab results submitted to the wrong
physician/doctors who did not request
for the lab test
Predictive Values
To assess the predictive value (PV) for a test, the sensitivity,
specificity, and prevalence of the disease in the population
being studied must be known.
The prevalence of the disease is the proportion
of population who has the disease. The incidence of a
disease is the number of subjects found to have the disease
within a defined period such a year , in a population of
100,000.
A positive predictive value (PV) for a test indicates
the number of patients with an abnormal test result
who have the disease, compared with all patients
with an abnormal result, as follows
Number of patients with disease
Positive PV = & with abnormal test results
Total number of patients with
abnormal test results
Positive PV
= ________Truepositives______________
True positives + False positives
A negative predictive value (PV) for a test indicates the
number of patients with a normal test result who do
not have the disease, compared with all patients with a
normal (negative) result, as follows
Negative PV = _ ___True negatives_________
True negatives + False negatives
Statistics
It is the science of gathering, analyzing,
interpreting and presenting data.
*mean – is a measure of central tendency . It is
associated with symmetrical or normal
distribution.
mean = Σx n
*Standard Deviation (SD) – is a measure of the
dispersion of values from the mean. It helps
describe the normal curve. A measure of the
distribution range the most frequently used
measure of variation.
* Coefficient of Variation (CV) is a percentile
expression of the mean; an index of precision.
CV = SD x 100
Mean
*Variance is called the standard deviation squared;
a measure of variability. It represents the difference
between each value and the average of the data.
V = (SD)2
Quality Control Chart
It is used to observe values of control materials over
time to determine reliability of the analytical
method.
It is utilized to observe and detect analytic errors
such as inaccuracy and imprecision.
1. Gaussian Curve (Bell-Shaped Curve)
*it occurs when the data set can be accurately
described by the SD and the mean.
*it is obtained by plotting the values from multiple
analyses of a sample.
*it is a population probability distribution that is
symmetric about the mean.
*it occurs when data elements are centered around
the mean with most elements close to the mean.
*it focuses on the distribution of errors from the
analytical method rather than the values from a
healthy or patient population.
*the total area under the curve is 1.0 or 100%
2. Cumulative Sum Graph (CUSUM)
*it calculate the difference between QC results *it easily identifies random and systematic errors.
and the target means.

*Common method: V mask

*it identifies consistent bias problems; it requires

computer implementation.

*this plot will give the earliest indication of

systematic errors (trend)and can be used with
the 135 rule.

*it is very sensitive to small, persistent errors that
commonly occur in the modern, low
calibration-frequency analyzer.
*results are out of control when the slope
exceeds 45o or a decision ± 2.7 SD IS
exceeded.
Errors which can be observed in L-J chart
a. Trend
it is formed by control values that either increase or
decrease for six consecutive days.
Main cause Deterioration of reagents

3. Youden/Twin Plot b. Shift

it is formed by control values that distribute
themselves on one side or either side of the mean for
*it is used to compare results obtained on a
six consecutive days.
high and low control serum from different
laboratories.
Shift in the reference range is due to transient
instrument differences.
*it displays the results of the analyses by plotting
the mean values for one specimen on the Main cause: Improper calibration of the instrument.
ordinate y-axis and the other specimen on the
abscissa x-axis.
c. Outliers

*the points falling from a center but on the 45o

*These are control values that are far from the main
line suggest a proportional error and points
set of values.
falling from the center but not on the 45o line
suggest a constant error.
*These are highly deviating values.
4. Shewhart Levey-Jennings Chart
*These are caused by random or systematic errors.
*it is the most widely used QC chart in the
clinical laboratory.
5. Westgard Control Chart
*it allows the laboratorians to apply multiple
rules without the aid of the computer. It recognizes that the use of simple upper and lower
control limits is not enough to identify analytical
*it is a graphic representation of the problems.
acceptable limits of variation in the results of an
analytical method. In Westgard, error detection rates can increase
without increasing the false rejection rate.
Westgard used the term control rule to indicate
if the analytical process is out of control.
Quality Control Chart
*Six Westgard rules are
typically used:
three are warning rules and
three are mandatory rules.
Lean System
It is a system for reducing waste (“nonvalued
activities”)especially in production or manufacturing
processes.
It was conceptualized to improve the automobile
industry in terms of the quality and efficiency of
automobile production.
It focuses on work flow actions in performing specific
tasks, procedures, or other activities accomplished
by critically reviewing each step in the process to
determine where inefficiencies can be eliminated.

A “Lean Clinical Laboratory” utilizes fewer resources,

reduces costs, enhances productivity, promotes
staff morale, and improves the quality of patient
care

Ex. Relocating analytic equipment to an area that

would require fewer steps, thus improving
turnaround time, consolidating test panels to fewer
instruments, eliminating the expense of maintaining
multiple instruments and supplies, accesability to
instruments and materials such as pipettes, culture
tablets, reagents, etc. and relocating staff to
maximize and minimize wasteful down time.
Westgard Rules

accept the results of the testing run, if only a

warning is violated.

reject the results of the testing run, when a

mandatory rule is violated.

increase the retesting range for a particular

assay, if either a warning or mandatory rule is
violated.

Six Sigma (6σ)

It measures the degree of variability or error in

products or services through statistics and
quantitative parameters – process defects or
errors analyzed, potential causes are identified
and improvements are implemented.

Main Goal: To reduce the number of defects to

near zero.
Carbohydrates
Blood Sugar
• Blood sugar or blood glucose
- Refers to sugar that is transported
through the bloodstream in order to
supply energy to all the cells in our bodies
- The sugar is made from the food we eat
- The human body regulates blood glucose
levels so that they are neither too high nor
too low
• Sugar
- Is a simple, crystalline, edible
carbohydrate and comes in a variety of
forms, all of the sweet
- Our body digests carbohydrates into
glucose, a simple sugar than can easily
convert to energy
- The chemical formula for glucose is C5H12O6
• The human digestive system breaks down the
carbohydrates from food into various sugar
molecules
– one of them is glucose, the body’s principal
source of energy. The glucose goes straight
from the digestive system into the
bloodstream after we have consumed and
digested food. Glucose can only enter cells if
there is insulin in the bloodstream too.
Without any insulin the cells would starve
• After we eat, blood sugar concentrations rise,
the pancreas releases insulin automatically so
that the glucose enters cells, as more and
more cells receive glucose, blood sugar levels
come down to normal again. Excess glucose is
stored as glycogen (stored glucose) in the liver
and muscles
• After a meal blood glucose levels rise, insulin
is released from the pancreas into the
bloodstream, blood sugar levels fallback
• If you have not eaten for a while and blood
glucose concentrations keep dropping, the
pancreas releases another hormone called
glucagon
• Glucagon
- Triggers the breakdown of glycogen into
glucose, thus pushing blood glucose
levels back up to normal
Causes of High Blood Sugar
Your blood sugar may rise
• Skip or forget your insulin or oral glucose
lowering medicine
• Eat too many grams of carbohydrates for the amount of
insulin you took, or eat too much carbs in general
• Have an infection
• Are ill
• Are under stress
• Become inactive or exercise less than usual
• Take part in strenuous physical activity, especially when
you blood sugar levels are high and insulin levels are low
Symptoms (Early Signs)
• Increased thirst
• Headache
• Trouble concentrating
• Blurred vision
• Frequent peeing
• Fatigue (weak, tired feeling)
• Weight loss
• Blood sugar more than 180mg/dl
How does the body control glucose in the blood?
• The normal blood glucose level ranges between 3.5-
7.8mmol/L.
• Insulin – lowers; Glucagon – heightens
The Glucose Journey (From food to fuel via bloodstream)
1. Glucose starts its journey as a carbohydrate food
2. When the food is eaten, it passes through the mouth,
stomach and small intestine
- All of these areas help to digest (break down) the
food to glucose
3. The glucose is absorbed from the small intestine into the
bloodstream
4. The bloodstream carries the glucose to its next stop, all
the body’s cells, particularly muscles, the brain and the
liver
- Your brain won’t function well if your glucose is low
5. The glucose can only enter the cells with the help of
insulin, a hormone which is made in the pancreas
6. As the blood glucose level rises after eating, the
pancreas releases insulin into the bloodstream. Insulin
travels to the cells, where it works to allow glucose to
enter the cells
7. Glucose is also directed to the liver to be stored for
later use. Between meals and overnight our body
can draw on the stored glucose for energy.
• During a fast, the blood glucose level is kept
constant by mobilizing the glycogen stores in the
liver
• During long fasts, gluconeogenesis is required to
maintain blood glucose levels because glycogen
stores are used up in about 24 to 48 hrs
• An individual with a fasting blood glucose level
- > 100mg/dL is referred to as hyperglycemic
• An individual with a fasting blood glucose level
- < 50mg/dL is referred to as hypoglycemic.
Hormones Affecting Blood Glucose
• Insulin
- Produced by the beta cells of the
pancreatic islets of Langerhans
- Promotes the entry of glucose into the
liver, muscle, and adipose tissue to be
stored as glycogen and fat
- Inhibits the release of glucose form the liver
• Somatostasin
- Synthesized by delta cells of the
pancreatic islets of Langerhans
- Inhibits secretion of insulin, glucagon,
and growth hormone, resulting in an
increase in plasma glucose level
• Growth Hormone and the
Adrenocorticotropic Hormone (Acth)
- Hormones secreted by an anterior pituitary
that arise blood glucose levels
• Cortisol
- Secreted by the adrenal glands
- Stimulates glycogenolysis, lipolysis and
gluconeogenesis
• Epinephrine
- Secreted by the medulla of the adrenal glands
- It stimulates glycogenolysis and lipolysis
- It inhibits secretion of insulin
- Physical or emotional stress causes
increased secretion of epinephrine and an
immediate increase in blood glucose levels
• Glucagon
- Secreted by the alpha cells of the
pancreatic islets of Langerhans
- Increases blood glucose by stimulating
glycogenolysis and gluconeogenesis
• Thyroxine
- Secreted by the thyroid gland
- Stimulates the glycogenolysis and
gluconeogenesis
- Increases glucose absorption from the intestines
Renal Threshold for Glucose
• Glucose is filtered by the glomeruli, reabsorbed by
the tubules, and normally not present in the urine
• If the blood glucose level is elevated, glucose
appears in the urine, a condition known as
glucosuria
• An individual’s renal threshold for glucose varies
between 160 and 180 mg/dl
- When blood glucose reaches this level or exceeds
it, the renal tubular transport mechanism
becomes saturated, which causes glucose to
be excreted into the urine.
• In severe hyperglycemia, one has polyuria,
ketonuria and glycosuria
- Polyuria – frequenturinating
- Ketonuria – ketone bodies in the urine
- Glycosuria – glycogen in the urine
• Ingestion of ethanol, salicylate and propanol may
cause hypoglycemia
Symptoms of Diabetes
• Always tired
• Frequent urination
• Sudden weight loss
• Wounds that won’t heal
• Sexual problems
• Always hungry
• Blurry vision
• Numb or tingling hands or feet
• Always thirsty
• Vaginal infections
Diabetes
• Describes a group of metabolic diseases in which the
person has high blood glucose (blood sugar), either
because insulin production is inadequate, or because the
body’s cells do not respond properly to insulin, or both
• Pancreas produce insulin according to the blood
glucose level
- Without the pancreas, a person can die
• Pathophysiology
- General
➢ Insulin – is the principal hormone that regulates
the uptake of glucose from the blood into cells of
the body, especially liver, adipose tissue and
muscle, except smooth muscle, in which insulin
acts via the IGF-1 (insulin-like growth factor-1)
➢ Therefore, deficiency of insulin or the
insensitivity of its receptors plays a central role
in all forms of diabetes mellitus
 ➢ The body obtains glucose from 3 main places:
o The intestinal absorption of food
o The breakdown of glycogen, the
storage form of glucose found in the
liver
o Gluconeogenesis, the generation of
glucose from non-carbohydrate
substrates in the body
➢ Insulin plays a critical role in balancing
glucose levels in the body:
o It can inhibit the breakdown of
glycogen or the process of
gluconeogenesis
o It can stimulate the transport of
glucose into fat and muscle cells
o It can stimulate the storage of glucose
in the form of glycogen
➢ Insulin is released into the blood by beta cells,
found in the islets of Langerhans in the
pancreas, in response to rising levels of
blood glucose, typically after eating
o Lower glucose levels result in
decreased insulin release from the beta
cells and results in the breakdown of
glycogen to glucose
o This process is mainly controlled by the
hormone glucagon, which acts in the
opposite manner to insulin
➢ If the amount of insulin available is
insufficient, if cells respond poorly to the
effects of insulin, if the insulin itself is
defective, then glucose will not be
absorbed properly by the body cells, the
net effect is persistently high levels of
blood glucose, poor protein synthesis, and
breakdown of fat storage – Acidosis
➢ When the glucose concentration in the
blood remains high over time, the kidneys
will reach a threshold of reabsorption ->
Glycosuria -> (this increases the osmotic
pressure of the urine) -> polyuria ->
(increased fluid loss) -> lost blood volume
will be replaced osmotically from water
held in body cells and other body
compartments -> dehydration ->
polydipsia
- Type 1 Diabetes Mellitus
➢ Type 1 Diabetes Mellitus is characterized
by loss of the insulin-producing beta cells
of the islets of Langerhans in the
pancreas, leading to insulin deficiency
➢ Glucose will go to the cell to release insulin
(it serves as the key for glucose to enter the
cell)
➢ This type can be further classified as
immune-mediated or idiopathic
➢ The majority of Type 1 Diabetes is of the
immune-mediated nature, in which a T-cell
mediated autoimmune attach leads to the loss of
beta cells and this insulin
o T-cell is the one attacking the beta cells that
produces insulin
➢ Most affected people are otherwise healthy
and of a healthy weight when onset occurs
➢ Sensitivity and responsiveness to insulin are
usually normal, especially in the early stages
➢ Type 1 Diabetes can affect children or adults,
but was traditionally termed “juvenile
diabetes” because a majority of these diabetes
cases were in children
➢ Type 1 Diabetes is partly inherited, with
multiple genes, including certain HLA
genotypes, known to influence the risk of
diabetes.
o Possibility to have diabetes so it is
somewhat a marker or receptor that you
are at risk of diabetes
➢ In genetically susceptible people, the onset of
diabetes can be triggered by one or more
environmental factors, such as a viral infection
or diet
➢ Among dietary factors, gluten may lead to type
1 diabetes, but the mechanism is not fully
understood
- Type 2 Diabetes Mellitus
➢ Type 2 DM is characterized by insulin
resistance
➢ The defective responsiveness of body tissues to
insulin is believed to involve the insulin
receptor
➢ In the early stage of type 2, the predominant
abnormality is reduced insulin sensitivity
➢ Type 2 DM is due to primarily to lifestyle
factors and genetics
- A number of lifestyle factors are known to be important
to the development of type 2 diabetes, including:
➢ Obesity
➢ Lack of physical activity
➢ Poor diet
➢ Stress
- Dietary factors also influence the risk of developing
type 2 diabetes such as:
➢ Sugar-sweetened drinks
➢ Type of fats in diet
o Saturated fats and trans fatty acids
 increasing the risk
o Polyunsaturated
and monounsaturated fat • Oral Manifestations andComplications
decreasing the risk - Periodontal changes are seen in Diabetes Mellitus
➢ Eating lots of white rice also may increase the - Salivary glands
risk of diabetes ➢ Xerostomia is common, but reason is unclear
➢ A lack of exercise is believed to cause 7% of ➢ Tenderness, pain and burning
cases sensation of tongue
- Gestational Diabetes - Dental caries
➢ Gestational Diabetes Mellitus (GDM) ➢ Increase caries prevalence in adult
resembles type 2 diabetes in several aspects with diabetes (xerostomia, increase
➢ Involves a combination of relatively saliva glucose)
inadequate insulin secretion and
responsiveness Feature Type 1 Type 2
➢ It occurs in about 2-10% of all pregnancies Diabetes Diabetes
Onset Sudden Gradual
and may improve or disappear after delivery
Age at onset Mostly in Mostly in adults
➢ However, after pregnancy approx. 5-10% of
children
women with gestational diabetes are found Body Size Thin or normal Often obese
to have diabetes mellitus, most commonly Ketoacidosis Common Rare
type2 Autoantibodies Usually present Absent
➢ Gestational diabetes is fully treatable, but Endogenous Low or absent Normal,
requires careful medical supervision Insulin decreased or
throughout the pregnancy increased
➢ Management may include dietary changes, Concordance in 50% 90%
identical twins
blood glucose monitoring, and in some Prevalence ~ 10% ~ 90%
cases, insulin may be required
➢ Hyperglycemia state shows a
➢ Through it may be transient, untreated
positive association with dental caries
gestational diabetes can damage the health
➢ Carious Lesion on teeth with Xerostomia.
of the fetus or mother
May cause secondary enlargement of
➢ Risks to the baby include:
parotid glands with Sialosis
o Macrosomia (high birth weight) – high
blood glucose levels in the mother
• Comparison of Type 1 and 2 Diabetes
Brings extra glucose to baby Causes
baby to put on extra weight
o Congenital heart defects How are Diabetes and Pre-Diabetes Diagnosed?
o Central nervous system abnormalities • Blood tests are used to diagnosis diabetes and
o Skeletal musclemalformations
pre- diabetes. Lab analysis of blood is needed to
➢ Increased levels of insulin in a fetus’ blood
ensure test results are accurate
may inhibit fetal surfactant production and
• Glucose measuring devices used in a health
cause respiratory distress syndrome
➢ A high blood bilirubin level may result from
care provider’s office, such as finger-stick
devices, are not accurate enough for diagnosis
RBC destruction (hemolysis) but may be used as a quick indicator of high
➢ In severe cases, perinatal death may occur, blood glucose
most commonly as a result of poor • Patients with Type 1 Diabetes will need to take
placental perfusion due to vascular insulin injections for the rest of their life. They
impairment must also ensure proper blood-glucose levels
➢ Labor induction may be indicated with by carrying out regular blood tests and
decreased placental function. following a special diet
• Type 1 Diabetes is usually diagnosed in children
➢ A Caesarean section may be performed
and young adults. Only 10% of people with
if there is marked fetal distress or an
diabetes have this form of the disease
increased risk of injury associated with • In type 1 diabetes, the body does not produce insulin
macrosomia, such as shoulder dystocia
 Types of Diabetes
• While type 1 and type 2 are the most
common form of diabetes, there are
others that you may hear about. Impaired
Glucose Metabolism or Pre-diabetes
• There are two pre-diabetes conditions:
- Impaired Glucose Tolerance (IGT)
is where blood glucose levels are
higher than normal but not high
enough to be classified as diabetes
- Impaired Fasting Glucose (IFG) is where
blood glucose levels are escalated in the
fasting state but not high enough to be
classified as diabetes
• On the other hand, despite eating so often and a lot,
the patients lose weight
• With the usage of protein as energy source, the
patient feels themselves tired and sluggish
- If blood glucose is too high, it is tried to be
thrown away by kidneys so the patients begin
to urinate so often
- As a result, the patients feel thirsty and start to
drink a lot
What happens if there is a problem with the production of
insulin?
• Glucose in blood is not able to go into the cell
• The cells can’t meet energy needs and energy is
tried to be provided from fat and protein
• Using fat as the energy source results the increasing
of ketone in the body
Urine Analysis
• Detection of Urinary Glucose (Glucosuria)
- First-line screening test for diabetes mellitus
- Normally, glucose does not appear in urine until
the plasma glucose rises above 160–180mg/dL
- In certain individuals due to low renal threshold
glucose may be present despite normal blood
glucose levels
- Conversely renal threshold increases with age
so many diabetics may not have glycosuria
despite high blood sugar levels
- Detection of glucosuria
➢ A specific and convenient method to
detect glucosuria is the paper strip
impregnated with glucose oxidase and a
chromogen system (Clinistix, Diastix),
which is sensitive to as little as 0.1% glucose
in urine
➢ Diastix can be directly applied to the urinary
stream, and differing color responses of the
indicator strip reflect glucose concentration
➢ Benedict’s and Fehling’s test can also detect
glucosuria
• Ketonuria
- Qualitative detection of ketone bodies can be
accomplished by nitroprusside tests (Acetest or
Ketostix), Rothera’s test, etc
- These tests do not detect Beta-hydroxyl butyric
acid, which lacks a ketone group
- Ketone bodies may be resent in a normal subject as a
result of simple prolonged fasting
• Microalbuminuria
- May be defined as an albumin excretion rate
intermediate between normality (2.5-25mg/day) and
macroalbuminuria (250mg/day)
- The small increase in urinary albumin excretion is not
detected by simple albumin stick tests and requires
confirmation by careful quantization in a 24hour
urine specimen
- The importance of microalbuminuria in the
diabetic patient is that it is a signal of early
reversible renal damage
- Performing an albumin-to-creatinine ratio is
probably easiest
- Microalbuminuria is a common finding (even at
diagnosis) in type 2 diabetes mellitus and is a risk
factor for macro vascular (especially coronary
heart) disease.
Blood Chemistry
• Blood Glucose Estimation
- Choice of Sample
➢ Plasma or serum from venous blood
samples has the advantage over whole
blood of providing values for glucose that
are independent of Hematocrit and that
reflect the glucose concentration to which
body tissues are exposed
➢ Plasma and serum are more readily
measured on automated equipment, they
are used in most laboratory
➢ If serum is used or plasma is collected from
tubes that lack an agent to block glucose
metabolism (such as fluoride), samples
should be refrigerated and separated
within 1 hour after collection
➢ The glucose concentration is 10-15% higher
in plasma or serum than in whole blood
because structural components of blood
are absent
- Fasting Blood Glucose
 ➢ Is measured after an overnight fast of
10 hours
➢ FBG estimation is better than random
blood glucose
o Normal: FPG < 5.6 mmol/L (100mg/dL)
o IFG: FPG = 5.6 to 6.9 mmol/L (100 to
125 mg/dL)
o Warrants the diagnosis of DM:
FPG >7.0 mmol/L (126mg/dL)
- Random Blood Glucose
➢ Random is defined as without regard
to time since the last meal
➢ RBG measurement is required only
during emergency
➢ The current criteria for the diagnosis
of DM emphasize that the FPG is the
most reliable and convenient test for
identifying DM in symptomatic
individuals
➢ A random plasma glucose concentration
>11.1 mmol/L (200 mg/dL)
accompanied by classic symptoms of
DM (polyuria, polydipsia, weight loss)
is sufficient for the diagnosis of DM
- Glucose Tolerance Test
➢ When the fasting plasma glucose level
is 125mg/dl or higher on more than
one occasion, further evaluation of
the patient with a glucose challenge is
unnecessary
➢ However, when fasting plasma glucose is
less than 126 mg/dl in suspected cases, a
standardized oral glucose tolerance test
may be done
➢ Methodology: 75g of glucose dissolved in
300mL of water is given after an overnight
fast to a person who has been receiving at
least 15-0-200g of carbohydrate daily for 3
days before the test
➢ Data Interpretation: The Diabetes Expert
Committee criteria for evaluating the
standard oral glucose tolerance test
Normal Impaired
Diabetes
glucose glucose
Mellitus
tolerance tolerance
Fasting plasma
glucose < 110 110 – 125 > 126
(mg/dL)
2 hrs after
< 140 140 – 199 > 200
glucose load
• Glycated Hemoglobin (HbA1C)Measurements
- HbA1C comprises 4-6% of total hemoglobin A1
- Since glycohemoglobin circulate within red blood
cells whose reflect the state of glycemia over the
preceding 8-12 weeks, thereby providing an
improved method of assessing diabetic control
➢ We will know the stored sugar in the body
within 3 months so they make use this fraction
of hemoglobin to monitor diabetes
➢ Monitored for 3 months
- Any condition that shortens erythrocyte survival or
decreases mean erythrocyte age (e.g., Recovery from
acute blood loss, hemolytic anemia) will falsely
lower HbA1C irrespective of the assay method used
- Methods for measuring HbA1C include:
➢ Electrophoresis
➢ Cation-exchange chromatography
➢ Boronate affinity chromatography
➢ Immunoassays
• Serum Fructosamine Estimation
- Serum fructosamine is formed bynonenzymatic
glycosylation of serum proteins (predominantly
albumin)
➢ Pregnant women make use of fructosamine
estimation
- Serum albumin has a much shorter half-life than
hemoglobin, serum fructosamine generally reflects
the state of glycemic control for only the preceding 1-
2 weeks
- Normal values vary in relation to the serum, albumin
concentration are 1.5-2.4 mmol/L when the serum
albumin level is 5g/dL
- When abnormal hemoglobin or hemolytic states
affect the interpretation of glycohemoglobin or
when a narrower time frame is required, such as for
ascertaining glycemic control at the time of
conception in a diabetic woman who has recently
become pregnant, serum fructosamine assays offer
some advantage
• Self-Monitoring of Blood Glucose
- Capillary blood glucose measurements
performed by patients themselves, as
outpatients, are extremely useful
- In type 1 patients in whom “tight” metabolic
control is attempted, they are indispensable
- There are several paper strips (glucose
oxidase, glucose dehydrogenase, or
hexokinase) methods for measuring glucose on
capillary blood samples
• Lipid Profile
- Serum total cholesterol is elevated
- Serum triglycerides are high
- Serum HDLc is low
 - Qualitative change in LDL particles,
producing a smaller dense particle whose
membrane carries supranormal amounts
of free cholesterol
- These smaller dense LDL particles are
more susceptible to oxidation, which
renders them more atherogenic
• Additional Tests
- The patient should be screened for DM-
associated conditions (e.g., Kidney, liver
and thyroid dysfunction)
- Individuals at high risk for cardiovascular
disease should be screened for
asymptomatic CAD by appropriate cardiac
stress testing, when indicated
- The classification of the type of DM may
be facilitated by laboratory assessments
• Serum insulin or C-peptide measurements do not
always distinguish type 1 from type 1 DM, but a low C-
peptide level confirms a patient’s need for insulin
Immunological Assays
• Antibodies to insulin, islet cells, or Glutamic
acid decarboxylase (GAD) can be estimated to
differentiate between the types of diabetes
mellitus
• They are absent in type 2 diabetes mellitus
• Latent autoimmune diabetes of adults, or
LADA, is a form of slow-onset type 1 diabetes
that occurs in middle-aged (usually white)
adults
• It can be differentiated from type 2 diabetes
by measuring anti-GAD65 antibodies
Management of Diabetes Mellitus
• Eliminate symptoms related to hyperglycemia
• Reduce or eliminate the long-term
microvascular and macrovascular
complications of DM
• Allow the patient to achieve as normal a
lifestyle as possible
• Management of Type 1 Diabetes Mellitus
- Because individuals with type 1 DM
partially or completely lack endogenous
insulin production, administration of
basal, exogenous insulin is essential for
regulating glycogen breakdown,
gluconeogenesis, lipolysis and
ketogenesis
- Likewise, insulin replacement for meals
should be appropriate for the
carbohydrate intake and promote normal
glucose utilization and storage
- Insulin Preparations
➢ Insulin is indicated for type 1 diabetes as well as
for type 2 diabetic patients with insulinopenia
whose hyperglycemia does not respond to diet
therapy either alone or combines with other
hypoglycemic drugs
➢ With the development of highly purified human
insulin preparations, immunogenicity has been
markedly reduced, thereby decreasing the
incidence of insulin allergy, immune insulin
resistance and localized lipoatrophy at the
injection site
- Methods of InsulinAdministration
➢ Insulin syringes and needle
o Plastic disposable syringes are available in
1mL, 0.5mL and 0.3mL sizes
➢ Insulin pen injector devices
o Insulin pens eliminate the need for carrying
insulin vials and syringes
➢ Insulin pumps
o Insulin infusion pumps are used for
subcutaneously delivery of insulin. These
pumps are small (about the size of a pager)
and very easy to program
o It is an alternative to multiple daily injections
of insulin by insulin syringe or an insulin pen
and allows for intensive insulin therapy
when used in conjunction with blood glucose
monitoring.
➢ Inhaled insulin
o A novel method for delivering a pre- prandial
powdered form of insulin by inhalation
(Exubera) has been approved by the FDA
o The FDA approved the first inhaled version
of insulin called Exubera from Pfizer Inc.
- Complications of InsulinTherapy
➢ Hypoglycemia, insulin allergy, immune insulin
resistance and lipodystrophy at the injection site
are some of the complications of insulin therapy
➢ Besides insulin therapy, lifelong dietary and
lifestyle modifications are required to be done to
achieve euglycemia
- Islet Cell Transplantation
➢ Is minimally invasive procedure and wide
application of this procedure for the treatment
of type 1 diabetes is limited by the dependence on
multiple donors and the requirement for potent
long-term immunotherapy
- Stem Cell Therapy
➢ Is one of the most promising treatments for the
near future. It is expected that this kind of therapy
can ameliorate or even reverse some diseases

• MANAGEMENT OF TYPE 2 DIABETES MELLITUS
- The goals of therapy for type 2 DM are similar
to those in type 1
- The care of individuals with type 2 DM
must also include attention to the
treatment of conditions associated with
type 2 DM (obesity, hypertension,
dyslipidemia, cardiovasculardisease)
- Detection or management of DM-related
complications
- Weight Reduction
➢ Treatment is directed toward
achieving weight reduction, and
prescribing a diet is only one means to
this end
➢ Behavior modification to achieve
adherence to the diet
➢ Increased physical activity to expend energy
– complications are micro and macro vascular
is also required
- Hypoglycemic Agents
➢ If the patient is not able to achieve
target glycemic control with weight
management and exercise, then
pharmacologic therapy is indicated
➢ Based on their mechanisms of action,
glucose- lowering agents are
subdivided into agents that increase
insulin secretion, reduce glucose
production and increase insulin
sensitivity
autoimmune destruction of insulin producing beta
cells of the pancreas
• The classical symptoms of type 1 DM are polyuria,
polydipsia, polyphagia and weight loss
• Is fatal unless treated with insulin
• Injection is the most common method of
administering insulin; insulin pumps and inhaled
insulin has been available at various times
• Diabetic keto-acidosis is the most common
complication of type 1 DM
Summary of Type 2 DM
• Is characterized by impaired insulin secretion,
insulin resistance, excessive hepatic glucose
production, and abnormal fat metabolism
• While many patients with type 2 diabetes present
with increased urination and thirst, many other
have an insidious onset of hyperglycemia and are
asymptomatic initially
• Hyperglycemic hyperosmolar state (HHS) is an
acute complication of type 2 DM. chronic
involving small and large blood vessels
respectively.
• Glucose lowering agents that either increase insulin
secretion, reduce glucose production, increase insulin
sensitivity, and enhance GLP-1 (Glucagon like peptide)
action are used to treat hyperglycemia
• The care of individuals with type 2 DM must also include
attention to the treatment of conditions associated
with type 2 DM (obesity, hypertension, dyslipidemia,
cardiovascular disease) and detection or management of
DM-related complications

• Biguanides
- Metformin is representative of this class of
agents
- It reduces hepatic glucose production
through an undefined mechanism and
improves peripheral glucose utilization
slightly
- Metformin reduces fasting plasma
glucose and insulin levels, improves the
lipid profile and promotes modest weight
loss
- The major toxicity of metformin, lactic
acidosis, can be prevented by careful
patient selection

Summary of Type 1 DM
• Type 1, IDDM, or juvenile diabetes – is a form
of diabetes mellitus that results from

Estimation of blood glucose
• Measurement of blood glucose is indicative of
current state of carbohydrate metabolism.
• Depending on time of collection:
o Fasting blood glucose-after an overnight
fast.
o Post meal or postprandial blood
glucose-2 hrs after the subject has taken
a normal meal.
o Random blood glucose - Any time of the
day.
Laboratory test for diagnosis
1. Estimation of blood glucose.
2. Oral glucose tolerance test.
Diet Plan
• Daily nutritional needs should be taken frequently
but small portions
Diabetes Myths
• People with diabetes should not exercise – NOT
TRUE!
- Exercise is important for people with diabetes,
as it is for everybody else
- Diabetic patients should discuss exercise with
their doctors before starting the exercise
• Fat people always develop type 2 diabetes
eventually – this is not true.
- Being overweight or obese raises the risk of
becoming diabetic, they are risk factors, but do
not mean that an obese person will definitely
become diabetic
• Children can outgrow diabetes – this is not true.
- Nearly all children with diabetes have type 1;
insulin-producing beta cells in the pancreas
have been destroyed. These never come back.
Children with type 1 diabetes will need to take insulin
for the rest of their lives, unless a cure in found one
day
• Only older people develop type 2 diabetes – things are
changing.
- A growing number of children and teenagers are
developing type 2 diabetes due to the explosion in
childhood obesity rates, poor diet, and physical
inactivity
• If you have diabetes, you cannot eat chocolates or
sweets – people with diabetes can eat chocolates and
sweets if they combine them with exercise or eat them as a
part of a healthy meal
• Diabetic patients cannot eat bread, potatoes or pasta –
people with diabetes can eat starchy foods. However,
they must keep an eye on the size of the portions
• Diabetes diets are different from other people’s – the
diet doctors recommend healthy nutrition; healthy for
everybody. Meals should contain plenty of vegetables,
fruit, whole grains, and they should be low in salt and
sugar, and saturated or trans-fat.
What can be done for diabetes at school?
• Brochures and films should be prepared to inform the
students about diabetes
• Students should be informed about the importance of
healthy eating and doing exercises
• School canteens should be controlled and warned
to sell healthy food and healthy drinks rather than
fast food and fizzy drinks
• Students should be informed about not eating fast
food
• Parents should be informed about healthy
nutrition and the importance of homemade food in
children’s bag
• Teachers should follow their students about their
health problems. If they have some symptoms with
any disease, they should contact the parents
• They should also inform the students about the
importance of their health.