Clinical Chemistry 1

LECTURE

MODULE 1 UNIT 2: CLINICAL LABORATORY APPARATUS AND SUPPLIES

I. Types of Glassware

A. Borosilcate
main glass-forming constituent: with silica and boron trioxide

a. Thermal properties b. Chemical properties

 ↓ thermal expansion  highly resistant to:
 ↓ thermal stresses under a given temperature  ↑ chemical resistance
gradient  is shown below.
 "Strain Point” – maximum safe operating ↓ RESISTANCE ↑ RESISTANCE
temperature of borosilicate glassware.  Minute scratching  water
 Examples: of glass surface  neutral and acid solutions
o Pyrex (Corning Glass Works, Corning, NY)  When heated  concentrated acids their mixtures
o Kimax (Kimble Glass Co., Vineland, NJ) above 500 C the  chlorine
↓ RESISTANCE ↑ RESISTANCE glass may acquire  bromine
 Minute  higher temperature gradients permanent  iodine
scratching of  sudden temperature changes stresses on cooling  organic matters
glass surface  thermal shocks  hydrofluoric acid  resists extended period of reaction
 When heated  very hot  resists temperatures above 100’C
above 500 C phosphoric acid  exceeds that of most metals and other materials
the glass may  alkaline solutions  withstand repeated dry and wet sterilization
acquire without surface deterioration and subsequent
permanent contamination
stresses on  Resistance to attack of various chemicals
cooling

c. Optical properties
 Laboratory glassware made from borosilicate glass show no noticeable absorption in the visible region of the spectrum. It appears
consequently clear and colourless.

B. Alumina-Silicate Glass D. Soda-lime glass

main glass-forming constituent: aluminum oxide
 It is similar to borosilicate glass but it has:
o greater chemical durability
o can withstand higher operating temperatures
o strengthened chemically rather thermally
 Characteristics of alumino silicate glass include:
-Higher maximum operating temperature than borosilicate glass
-Particularly suitable for use as a gauge glass
-High silica content
-Comparable to fused quartz in its heat resistance, chemical
stability, and electrical characteristics
-Strengthened chemically rather than thermally
-Used for high-precision analytical work
-Radiation-resistant and can also be used for optical reflectors and
mirrors
C. Vycor glass
 main glass-forming constituent: 96% silica
 has several exceptional properties
 similar to fused quartz in its thermal properties
 may be used at much higher temperatures than borosilicate glass
 This glassware can be used continuously at 9000C, and
 known for its excellent chemical and physical properties
 products that resist the impact of chemical medium and
temperature differences momentarily and limitedly are mostly
manufactured of the soda-lime glass
 lower energy demand and longer working times make the soda-
lime glass the most inexpensive type of glass to manufacture
laboratory glassware
 can be recycled easily
 used to make pipettes
E. Low actinic glass
 is tinted dark brown or amber
 can be created from any material
 named after the colour
 done to protect light-sensitive chemical compounds from getting
altered by infrared radiation, visible light and ultraviolet radiation
 are of high thermal resistance with a red color
 density of the red color is adjusted to permit adequate visibility of
contents yet give maximum protection for light sensitive materials
such as bilirubin standards

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Clinical Chemistry 1
LECTURE

MODULE 1 UNIT 2: CLINICAL LABORATORY APPARATUS AND SUPPLIES

intermittently to 12000C.
It is utilized for high thermal, drastic heat shock and extreme
chemical treatment with acids (except hydrofluoric) and dilute
alkali.

II. Types of Plasticware

ADVANTAGES DISADVANTAGES
a. Less expensive and more durable than glassware a. Leaching of surface-bound constituents into solutions
b. Unbreakable b. Permeability to water vapour
c. Preferred for certain analyses in which glass can be damaged by c. Some evaporation through breathing of the plastic
chemicals used in the testing. (E.g. Generally, alkali solutions must d. Absorption of dyes, stains, or proteins
be stored in plastic) e. Cannot be used for HPLC (Solvents readly attack plasticware)

Polysterene Polyethylene Polypropylene Tygon Teflon

CPE LPE
Temperature Limit 70 80 130 135 95 205
(0C)
Transparency Clear Translucent Opaque Translucent Translucent Clear to Translucent
Autoclavable No No With Yes Yes Yes
caution
Flexibility Rigid Excellent Rigid Rigid Excellent Excellent (But easily
scratched and
warped)
Usage Sample Disposable All-purpose reagent Screw-cap closure Tubings Stopcocks, Wash
plasticware bottles, Plastic test tube bottles bottles, Beakers (for
rack, Carboys, cryogenic
Droppers, Specimen experiments)
transport containers,
reagent bottles
Chemical resistance Useful with water They have excellent Has the same -- This resin possesses
and aqueous salt chemical resistance to chemical resistance excellent chemical
solutions. It is most substances, with as LPE resistance to almost
recommended for the exception of all chemicals used in
use with acids, aldehydes, amines, the clinical
aldehydes, ketones, ethers, hydrocarbons, laboratory. With
ethers, and essential oils. For unique anti-adhesive
hydrocarbons, or CPE, exceptions would properties and non-
essential oils. include lubricating oils wettable surface.
Alcohols and bases and silicones. The usage Bottles and beakers
can be used, but of any of the above- out of this is suitable
storage beyond 24 named chemical groups for cryogenic
hours is discouraged. should be limited to 24 experiments. Resists
hours at room extreme
temperature temperatures (-
2700C to 2550C)

Polycarbonate Polyallomer Polyallomer Polysulfone

-- **For tubing PVC
Temperature Limit 135 70 130 165
(0C)
Transparency Very clear and shatterproof Clear -- Translucent Clear
Autoclavable Yes No Yes Yes Yes
Flexibility Rigid (Sterilizing reduces Rigid Flexible Moderate Rigid
mechanical strength)
Usage Sample All-purpose large reagent Bottles Tubing -- --
containers, carboys, plastic
test tube rack, graduated

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Clinical Chemistry 1
LECTURE

MODULE 1 UNIT 2: CLINICAL LABORATORY APPARATUS AND SUPPLIES

cylinders, and centrifuge tubes

Chemical resistance Very susceptible to damage by -- -- -- --
most chemicals. It is resistant
to water, aqueous salts, food,
and inorganic acids for a long
period of time.

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Clinical Chemistry 1
LECTURE

MODULE 1 UNIT 2: CLINICAL LABORATORY APPARATUS AND SUPPLIES

III. Types of Reagent

CHEMICALS ORGANIC REAGENTS

 Analytic chemicals exist in varying grades of purity:  Organic reagents also have
o analytic reagent (AR) varying grades of purity that
o ultrapure differ from those used to classify
o chemically pure (CP) inorganic reagents.
o United States Pharmacopeia (USP)  These grades include a practical
o National Formulary (NF) grade with some impurities;
o technical or commercial grade. o CP, which approaches
 A committee of the American Chemical Society (ACS) established specifications for AR grade the purity level of
chemicals, and chemical manufacturers will either meet or exceed these requirements. reagent grade
 Labels on reagents state the actual impurities for each chemical lot or list the maximum allowable chemicals
impurities. o spectroscopic
 The labels should be clearly printed with the percentage of impurities present and either the initials (spectrally pure)
AR or ACS or the term o chromatographic
 For laboratory use or ACS Standard-Grade Reference Materials. grade organic
 Chemicals of this category are suitable for use in most analytic laboratory procedures. reagents, with purity
 Ultrapure chemicals have been put through additional purification steps for use in specific procedures levels attained by their
such as chromatography, atomic absorption, immunoassays, molecular diagnostics, standardization, respective procedures
or other techniques that require extremely pure chemicals. o reagent grade (ACS),
 These reagents may carry designations of HPLC (high-performance liquid chromatography) or which is certified to
chromatographic on their labels. contain impurities
 Because USP and NF grade chemicals are used to manufacture drugs, the limitations established for below certain levels
this group of chemicals are based only on the criterion of not being injurious to individuals. established by the
 Chemicals in this group may be pure enough for use in most chemical procedures; however, it should ACS. As in any analytic
be recognized that the purity standards are not based on the needs of the laboratory and, therefore, method, the desired
may or may not meet all assay requirements. organic reagent purity
 Reagent designations of CP or pure grade indicate that the impurity limitations are not stated and is dictated by the
that preparation of these chemicals is not uniform. particular application.
 It is not recommended that clinical laboratories use these chemicals for reagent preparation unless
further purification or a reagent blank is included.
 Technical or commercial grade reagents are used primarily in manufacturing and should never be
used in the clinical laboratory.
 Analytical reagent grade chemicals state the actual impurities for each lot.

A. Types of Chemicals

Analytic reagent (AR) grade Ultrapure chemicals USP and NF Chemically pure Technical
(CP) / Pure Grade or
chemicals commercial
grade
 Are of very high purity and are suitable for  Have been put  Used to  Aka Less pure  Used
use in most analytical procedures through additional manufacture grade chemicals primar
 Specifications for AR grade chemicals are purification steps drugs  Impurity ily in
established by the American Chemical  Uses:  Only limitation limitations are manuf
Society (ACS) chromatography, established for not stated acturin
 Labels should state the actual impurities atomic absorption this group is not  Preparation of g and
for each chemical lot or list the maximum immunoassays, being injurious to these chemicals should
allowable impurities Label: molecular diagnostics, individuals is not uniform - never
o Clearly printed with the standardization, or  Purity standards Not be
percentage of impurities present other techniques that are not based on recommended used in
o Should have the initials AR or require extremely the needs of the for reagent the
ACS pure chemicals laboratory, preparation clinical
o Should have the term For  Labels may carry therefore, may or unless further laborat
laboratory use or Standard- designations of ‘HPLC’ may not meet all purification or a ory
Grade Reference Materials or ‘chromatographic’ assay reagent blank is

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Clinical Chemistry 1
LECTURE
A. Spectroscopic grade organic
reagents
“Spectrally pure” Purity levels
attained by spectrophotometric
procedures
A. Primary standard
A primary standard is a highly
purified chemical that can be
measured directly to produce a
substance of exact known
concentration and purity. The ACS
has purity tolerances for primary
standards, because most biologic
constituents are unavailable within
these tolerance limitations.
Highly purified chemical
-Can be measured directly to
produce a substance of EXACT
known concentration and purity
-ACS purity tolerance: 100 + 0.02%
-Most biological constituents are
unavailable within these limitations
(Physiologic primary standards are
generally unavailable)
MODULE 1 UNIT 2: CLINICAL LABORATORY APPARATUS AND SUPPLIES
requirements included
B. Types of Organic Reagents
B. Chromatographic grade C. Reagent grade (ACS) D. Chemically pure (CP)
organic reagents
Minimum purity of 99% Certified to contain impurities Approaches the purity level of
determined by gas below certain levels established reagent grade chemicals
chromatography by the ACS
C. Reference materials
B. Standard Reference materials (SRM) C. Secondary standard
Standard Reference Materials are used as primary standard in the A secondary standard is a
clinical laboratory. They are relatively expensive and should be used as substance of lower purity
a reference for the comparison of commercially obtained standards whose concentration is
and reagents. determined by comparison to
a primary standard
-Developed by the National Institute of Standards and Technology -Substance of lower purity
(NIST) specifically for use in the clinical chemistry laboratory -Concentration is determined
-Assigned a value after careful analysis and has certified chemical by comparison with a primary
composition standard
- May not possess the purity equivalent of an ACS primary standard -Depends not only on its
but has been characterized for a certain chemical or physical property composition, which cannot be
-Used in place of an ACS primary standard in clinical work and often directly determined, but also
used to verify calibration or accuracy/ bias assessments on the analytic reference
-Used by many manufacturers when producing calibrator and method
standards -Manufacturers are required
* ”Traceable to NIST” to list the SRM or primary
- Calibrator and standard that are manufactured using/ basing from standard used for comparison
NIST SRM
CAMACHO, ARRIANE CYREL A. | BSMLS 2
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