CLINICAL CHEMISTRY

• one of the busiest sections of the clinical laboratory

• measurement of the amount of chemicals or quantitative analysis of
metabolites in body fluids, particularly blood.

Clinicians request for the test:

MLS/ CLS

• accurately perform the rest

• ensure valid results

MLS/ CLS must:

• understand the principle of the tests

• understand the physiology of the body system involved
• know how to interpret the result
• realize the medical use/ relevance of the test
CONVERSIONS

INACCURATE PROCEDURE: • Smaller to larger = move decimal to the LEFT.

o Ex: 10mg = 0.0 0 0 0 1 0 = 0.00001 kg (or 1 x 10 -5 kg)
• oversight of a disease • Larger to smaller = move decimal to the RIGHT.
• mismanagement of the patient o Ex: 1kg = 1 0 0 0 0 0 0 . = 1,000,000 mg (or 1 x 10 +6 mg)

Test may be done: • Solution components are almost always expressed as homogenous mixture
of 2 or more solutions
• individually
• panels CONCENTRATIONS
o group of individual tests
▪ related to each other (belonging to one organ) = the amount or quantity of solute per volume or in a given quantity of
▪ or which demonstrate multisystem involvement solution / solvent.
▪ which screen for the specific cause among diseases
• Solute = when it is dissolved in a solution
with shared or common presentations
• solvent = dissolving matrix

> two most common expression of concentrations are based on:

CLINICAL LABORATORY CALCULATIONS & MATHEMATICS a) molarity
b) concentration based on %
Metric System
1) % w/v (weight/volume) solid/liquid
• most commonly used in scientific measurements 2) % v/v (volume/volume) liquid/liquid
• decimal based system 3) % w/w (weight/weight) w of solution

If solutions are not prepared accurately:

a) experiments may fail DEFINITIONS:

b) a waste of time and money.
1. Avogadro's number: It is generally defined as 6.022 x 1023 molecules of a
Biological experiments are particularly sensitive to alterations in chemical substance, and is the number of molecules in one mole of a substance.
composition of solutions.
= forms the basis of the molarity system
1) enzymatic reactions are extremely sensitive to alterations in pH,
2. The gram-molecular weight (as depicted in the Periodic Table of Elements)
and salt concentrations;
is the weight (in grams) of one mole of a substance.
2) incorrectly prepared media may inhibit bacterial growth;
3) DNA migration in agarose gels can be altered by inexact
calculation.
MOLARITY
= units of measurement of molarity
STANDARD METRIC UNITS FOR EXPRESSION:
moles/liter = mol/L or M
• weight = gram
• length = meter • 1 mole of a compound = gram molecular weight of that compound
• volume = liter
• time = second = obtained by adding the atomic weights of the atoms comprising the
compound
The metric System has the advantage that fractional parts of measurements
can be derived by multiplying the standard by some power of 10
 BASE OR SALTS
MOLALITY
= equivalent weight is the quantity of substance that will react with one
• expression of concentration that differs from molarity replaceable hydrogen.
• designates the amount of solute per 1000g (1kg) of solvent rather
than final solution as used in molarity
• weight / weight measurement rather than weight / volume, and is
thus independent of temperature variation, making it more
accurate concentration measurement than molarity.
• units of measurement of molality:
o moles/1000g
• less convenient to work with, and is not commonly used in clinical
laboratory.
• Since most solutions used in clinical laboratory are aqueous
solutions, there is very little difference between molality &
molarity.
NORMALITY

• the number of equivalent weights (or equivalents) of solute per

liter of solution,
• units of measurement:
o equivalents /liter
• expressed as N

CONCENTRATION BASED ON %

HYDRATES:

NORMALITY • Water of hydration

• water molecules attached to each molecule of salt, in

• similar to molarity except that concentration is based on
manufactured chemical compound.
equivalent weight instead of molecular weight
o anhydrous = without water
EQUIVALENT WEIGHT o monohydrate = 1 H2O
o pentahydrate = 5 H2O
• the mass of an element or compound that will combine with or • Ex. of prescribed hydrate of salt: copper sulfate
replace 1 mole of hydrogen (H). o CuS04 = 160g
• dependent on the total charge of the positive ion, or the valence o CuS04 . H2O = 178g
of the element. o CuS04 . 5 H2O = 250g

ACID COMPOUND

= an equivalent is the quantity of substance that contain one replaceable

hydrogen.

DILUTIONS

• weaker solution is made from stronger solution,

• diluent, such as water, added to an aliquot of a stronger solution,
to produce lesser concentration.
 • A 1:10 dilution can also be expressed as = 1 to 10 or 1/10

which means:

Examples:

• Another formula for dilution: V1 x C1 = V2 x C2

• use to know what volume or concentration of one substance will
neutralize another known substance.

SERIAL DILUTIONS

• titering
• useful in serologic tests when estimates of the volume of antibody
is necessary.
• “Dilution fold” is constant in all tubes
• volume transferred is constant to each successive tube.

• total volume = volume being transferred + volume of diluent

already in the tube.
• the last volume to be transferred is discarded from the last tube.

TEMPERATURE: (3 scales)

Water Celsius C Fahrenheit F Kelvin K

Freezing point 0C 32 F 273.15 K

Boiling point 100 C 212 F 373.15 K
Number of scales 100 units 180 units Same C

SPECIFIC GRAVITY

• used when working with concentrated acids or base

KELVIN
• used for very high or very low temp.
• an absolute, thermodynamic temperature scale using as its null
point absolute zero, below which temperatures do not exist.
• the weight of 1 ml of any liquid
• method of measuring density
• ratio of mass / volume [g/ml or ml/g]
• concentrated commercial liquids: check labels for specific gravity
& percent purity (assay)
 INTERNATIONAL SYSTEM OF UNITS

• Systeme International d’Unites (SI), adopted in 1960

• established, whereby all quantitative measurements could be
expressed in standard units, although conventional Metric System
are still frequently used today.

RADIOISOTOPES

• Half life – time required for a given amount of radioactivity to

decrease to one-half its original value
o The amount of radioactivity decreases by a factor of 2
for every half-life period
o If an isotope has a half-life of 8 hours and an activity of
16 mCi (millicuries), its activity will drop to 2 mCi in 24
hours.
• 1 mCi activity = 3.7 X 107 disintegration/second (1000 mCi = 1 Ci)

ELEPHANT FORMULARY

IONIC STRENGTH • Conversion of Conventional Chemistry Units to SI Units

• Bicarbonate, Chloride, CO2 (total), Potassium, & Sodium do not
• First step in calculations is calculation of so-called ionic strength, require conversion from conventional to SI units.
using following formula:
Albumin g/dl x 10 = g/L
Ammonia mg/dl x 0.5872 = mmol/L

Bilirubin mg/dl x 17.10 = mmol/L

• where Ci is a molar concentration of ith ion present in the solution
Calcium mg/dl x 0.2495 = mmol/L
and zi is its charge. Summation is done for all charged molecules
present in the solution.
 Cholesterol mg/dl x 0.02586 = mmol/L

CO2 pressure, pCO2 mmHg x 0.1333 = kPa

Creatinine mg/dl x 88.4 = µmol/L

Glucose mg/dl x 0.05551 = mmol/L

Lactate mg/dl x 0.111 = mmol/L

Magnesium mg/dl x 0.4114 = mmol/L

O2 pressure, PO2 mmHg x 0.1333 = kPa

Phosphorus mg/dl x 0.3229 = mmol/L

Protein g/dl x 10 = g/L

Urea Nitrogen mg/dl x 0.7140 = mmol/L

To convert blood glucose readings between the two units:

• Divide a mg/dL figure by 18 (or multiply by 0.055) to get mmol/L.

• Multiply a mmol/L figure by 18 (or divide by 0.055) to get mg/d

How to convert meq/L in to mg/L?

• You must multiply meq/L by mg/meq to get mg/L.

• mg/meq is simply the inverse of the equivalent weight.

CALCULATING pH
PIPETS/ PIPETTES
• pH is only meaningful when applied to aqueous (water-based)
solutions. To calculate the pH of an aqueous solution you need to • used to transfer or measure aliquots of a liquid.
know the concentration of the hydronium ion in moles per liter • clinical laboratorian should be aware when selecting type of pipet
(molarity). The pH is then calculated using the expression: to use
• “Mouth pipeting” should not be done
o pH = - log [H3O+]. • Accessories:
▪ pipet aid
• Ex: Find the pH of a 0.0025 M HCl solution. The HCl is a strong acid ▪ pipet bulb
and is 100% ionized in water. The hydronium ion concentration is ▪ pipet tips
0.0025 M. Thus:
According to the NBS (National Bureau of Standards), Class A pipets are more
o pH = - log (0.0025) = - (- 2.60) = 2.60 accurate than Class B pipets. In the clinical chemistry laboratory, the
o [press 0.0025; press log; then press equal sign] volumetric glassware must all be Class A.

Pipets may be classified as to their:

• Design - (To Contain or To Deliver)

CALCULATING THE HYDRONIUM ION CONCENTRATION FROM pH
• The hydronium ion concentration can be found from the pH by the reverse
of the mathematical operation employed to find the pH.
[H3O+] = 10-pH or [H3O+] = antilog (- pH)
TO CONTAIN PIPETS.
Example: What is the hydronium ion concentration in a solution that has a pH
of 8.34?
• Drainage Characteristics - (Blow-out or Self-draining)
• Function - (Measuring/Graduated or Transfer)
o M/G - e.g., serologic, Mohr, bacteriologic, Ball-Kolmer-
Kah, micropipette;
o T - e.g., volumetric, Ostwald-Folin, Pasteur pipets,
automated macro- or micropipet).
• These are also called rinse-out pipets. They contain an exact
amount of liquid which must be completely transferred for
accurate measurement.
• Examples are Micro-Folin, dual purpose, Sahli hemoglobin, Kirk
Micro, WhiteBlack lambda, transfer micro, measuring micro, and
Lang-Levy. None of these met the
• Class A specification of NBS.
• calibrated by introducing the exact wt of Hg equivalent to the
aqueous volume desired
TO DELIVER-BLOW-OUT PIPETS. 2. Disposable capillary micropipette:

• They transfer or deliver an exact amount of the liquid and are not
rinsed out.
• Examples are Ostwald-Folin (recognized by its bulb), serological
pipets, serological long tip, and serological large tip.
• These pipets are readily identified by the two forested or etched
bands near the mouthpiece. The diameter of the pipets is uniform
and their volume graduations extend up to the delivery tip. The
last blown-out drop of the liquid is included in the delivered
volume.
TO DELIVER-NON-BLOW-OUT PIPETS.
• These pipets are filled and allowed to drain by gravity. The pipets
must be held vertically and the tip placed against the side of the
accepting vessel.
• Examples are as follows:
o Volumetric pipets:
▪ These are the most accurate. They are Class
A pipets and can be used for diluting
standards, calibrators or quality control
materials.
▪ They hold and deliver specific volumes
indicated at the upper end of the pipet.
After draining, the amount transferred is
equal to the stated value.
o Mohr pipets:
▪ These are also Class A pipets. Their
graduations are made at uniform intervals
but well away from the tapered delivery tip.
These pipets are calibrated to deliver-in
between.
▪ The listed accuracy is for the full volume;
the smaller the volume used, the less
accurate the volume delivered. Note that
these pipets are never blown out since they
deliver volumes point to point.
• it consists of capillary tubing with a line marking a specific volume.
It is filled to the line by capillary action. The liquid is delivered by
positive pressure (blowout) through a medicine dropper or an
equivalent device. It is calibrated to contain therefore it requires
rinsing.
3. Pasteur (glass) & Disposable (plastic) Transfer Pipets
• used to transfer liquids from 1 receptacle to another
• may be used to remove serum from a clot tube or plasma from
anti-coagulated tube
4. Semi-automated Pipets
• single or multichanneled
• no pipetting bulb nor washing needed
• a plunger or trigger is used to aspirate the liquid into the pipet
• techniques used:
o Air-Displacement Techniques
▪ uses suction to draw up the fluid
o Positive Displacement
▪ uses a mechanical device such as piston to
displace the liquid to be drawn up.
▪ like the movement of the barrel in
hypodermic syringe
• use plastic tips usually polypropylene (retain < film)
• e.g. Eppendorf pipets
• Electronic Pipetters provide programmable application
parameters:
o pipetting mode:
▪ blowout, manual, rinsing
o fixed-volume mode:
o dispensing mode
▪ provides repetitive dispensing of constant
volume.

DISPENSERS

• used to add repeated volumes of reagent or diluent to serial

solutions or receptacles.
• consist of a reagent bottle to which a plunger with a valve system
is attached.

MICROPIPETS.
• They contain or deliver volumes ranging from 1 to 500 µL. they
are also called lambda pipets. A lambda is equal to a volume of 1
µL. Examples of micropipets are as follows:

1. Lang-Levy pipet:

• It is the original micropipette. It is filled to the constriction.

Probably, it is the most accurate for micro-volumes, but no longer
commonly used today.