1ST SEMESTER
Clinical chemistry 1 / LECTURE
Overview and lab mathematics
INTRODUCTION TO CLINICAL CHEMISTRY
Clinical Greek word “Kline” meaning bed.
Chemistry The science that deals with the
elements, their compounds and the
chemical structure and interaction of
matter.
Clinical Chemistry Is the area of chemistry that is generally
concerned with quantitative and
qualitative analysis of bodily fluids for
diagnostic and therapeutic purposes.
• The discipline originated in the late 19th century with the use of
simple chemical reaction tests for various components of blood
and urine.
• In the many decades since, other techniques have been applied
as science and technology have advanced, including the use
and measurement of Enzyme Activities, Spectrophotometry,
Electrophoresis, and Immunoassay.
• There are now many blood tests and clinical urine tests with
extensive diagnostic capabilities.
CLINICAL CHEMISTRY
• A quantitative science that is concerned with measurement of
amounts of biologically important substances (called analytes) in
body fluids.
• A science, a service, and an industry.
- AS A SCIENCE, it links the knowledge of general
chemistry, organic chemistry, and biochemistry with an
understanding of human physiology.
- AS A SERVICE, it produces objective evidence from which
medical decisions may be made.
- AS AN INDUSTRY, clinical laboratories are businesses,
which operate under the regulations and practices.
• Generally, the primary purpose of doing clinical chemistry is to
use it to perform analytic procedures that yield accurate and
precise information; and that will aid in the patient diagnosis and
treatment.
COMMON ANALYTES IN THE
CLINICAL CHEMISTRY LABORATORY
• Ions, Salts, and Minerals
- Sodium, Potassium, Calcium, Chloride, CO2, Lead, Iron
• Small Organic Molecules
- Metabolites (Glucose, Cholesterol, Uric Acid)
- Therapeutic Drugs (Vancomycin, Theophylline, Digoxin)
- Toxicology (Alcohol, Salicylate, Acetaminophen)
- Drugs of Abuse (Cocaine, Barbiturates)
• Large Molecules
- Transport Proteins (Albumin, Transferrin, Haptoglobin)
- Enzymes (Enzymes, Amylase, Creatinine, Kinase)
- Specific Proteins (Immunoglobulins, C-reactive proteins,
Complement)
- Diabetes Marker (HbA1c or Hemoglobin A1c)
FLUIDS TYPICALLY USED FOR
CLINICAL CHEMISTRY TESTS
• Blood (main – serum; whole blood for HbA1c), Urine,
Cerebrospinal Fluid, Amniotic Fluid, Saliva, Synovial Fluid,
Pleural Fluid, Pericardial Fluid, & Peritoneal Fluid
REGULATORY GUIDELINES THAT DEFINE THE
PRACTICE OF CLINICAL CHEMISTRY
• Clinical Laboratory Improvement Amendments (CLIA) of
1988: quality standards for all clinical laboratories to ensure
accuracy, reliability and timeliness of patient test results
regardless of where the test was performed
• Two Levels of Regulations
a. Waived Tests
o simple laboratory examinations and procedures that
are cleared by the U.S. Food and Drug Administration
(FDA) for home use. (ex. pregnancy test)
b. Nonwaived Tests
o moderately and highly complex tests as defined by the
requirements for operator skill, reagent preparation,
SY 2023-2024 CCHM321
automation and the difficulty of interpretation of
results.
o these are regulated under guidelines that cover quality
standards for proficiency testing (PT), patient test
management, quality control, personnel qualifications,
and quality assurance.
** Proficiency testing (PT) is a method of monitoring
accurate outcome. There are test samples coming
from outside source that will be analyzed and results
will be compared to those of reference laboratories
and scored for accuracy.
• Occupational Safety and Health Act (OSHA): provide
guidelines for safe operation of testing processes regulations
include guidelines for operating safety equipment and
identifying, handling and storing chemical hazards.
PURPOSE OF CLINICAL CHEMISTRY
MLS / MEDICAL TECHNOLOGIST
• Perform analytical procedures that yields accurate and precise
information, aiding in patient diagnosis and treatment.
• Correctly use basic supplies and equipment.
• Possesses an understanding of fundamental concepts critical to
any analytic procedure.
SCOPE OF PRACTICE:
General Services Provided by
Clinical Laboratory Scientists / Medical Technologists
1. Assuring reliable test results that contribute to the
prevention, diagnosis, prognosis, and treatment of
physiological and pathological conditions.
• This assurance requires:
A. Producing accurate test results. (through PT)
B. Correlating and interpreting test data.
C. Assessing and improving existing laboratory test
methods.
D. Designing, evaluating, and implementing new
methods.
** C and D go hand in hand.
2. Designing and implementing cost-effective administrative
procedures for laboratories, including their services and
personnel.
3. Designing, implementing, and evaluating processes for
education and continued education of laboratory personnel.
4. Developing and monitoring a quality assurance system to
include:
A. Quality control of services.
B. Competence assurance of personnel.
5. Promoting an awareness and understanding of the services
they render to the consumer, the public, and other health-
care professionals.
LABORATORY MATHEMATICS
• UNIT OF MEASUREMENT
1. Number related to the actual test value
2. A label identifying the units
**Ex. 85 mg/dL – 85(number/value); mg/dL (label/unit)
INTERNATIONAL SYSTEM OF UNITS (SI)
• Adopted internationally in 1960.
• Preferred in scientific literature and clinical laboratories.
• Standard method of describing physical quantities.
• Based on the metric system.
(Ex. kilo, hecto, deca, gram, milli, micro, nano, pico)
• The only system employed in many countries.
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Cchm321 LEC 1st SEMESTER | overview and lab mathematics
• Basic Unit REPORTING OF REFERENCE RANGES
Base Quantity Name Symbol • Newborn Infant
Length Meter m • Infants
Mass Kilogram kg • Pediatric
Time Second s
Electric current Ampere A IMPORTANT NOTE: ROUND OFF ALL ANSWERS TO THE
Thermodynamic Kelvin K NEAREST TENTHS, MEANING ONLY ONE (1) DECIMAL PLACE.
temperature ADD ZERO IF WHOLE NUMBER **21.0
Amount of Mole mol
substance PERCENT SOLUTION
Luminous intensity Candela cd • Parts per 100

• Derived Unit PERCENT BY MASS (mass-mass percent or %w/w)

Base Quantity Name Symbol Is the mass of solute in a solution divided by the total mass of
Selected Derived solution, multiplied by 100 (to put the value in terms of
Frequency Hertz Hz percentage).
Force Newton N
Celsius Degree Celsius °C w/w% = mass of solute x100
temperature mass of solution -------> mass of solute + mass of solvent
Catalytic activity Katal kat
** derivative or a mathematical function describing one of the PERCENT BY VOLUME (volume-volume percent or %v/v)
basic units Is the volume of the solute in a solution divided by the total
volume of solution, multiplied by 100.
• Non-SI
Base Quantity Name Symbol v/v% = volume of solute x100
volume of solution -------> volume solute + volume solvent
Selected Accepted Non-SI
Minute (time) (60 s) min
MASS-VOLUME PERCENT (%w/v)
Hour (3,600 s) h
Is the mass of solute in a solution (in grams) divided by the
Day (86,400 s) D
total volume of solution (in milliliters), multiplied by 100.
Liter (volume) (1 dm3 = 10-3 m3) L
Angstrom (0.1 nm = 10-10 m) A w/v% = mass of solute x100
** also includes gram and plane angles volume of solution (mL) -> volume solute + volume solvent

SIGNIFICANT FIGURES
• Minimum number of digits needed to express a particular value PERCENT SOLUTIONN – weight / weight
in a scientific notation without losing its accuracy.
• RULES: • Problem 1.
1. All nonzero numbers are significant. A saline solution with a mass of 355 g has 36.5 g of NaCl
814.2 = 4 dissolved in it. What is the mass/mass percent concentration of
2. All zeros between nonzero numbers are significant. the solution?
1001 = 4 36.5 g_ x100 = 10.3%
3. All zeros to the right of the decimal are not significant when 355 g
followed by a nonzero number.
0.000641 = 3 • Problem 2.
4. All zeros to the left of the decimal are not significant. What is the percent by mass of rubbing alcohol in a solution that
(leading zeros – aka “place holders”) contains 275 g of rubbing alcohol in 500 g of solution?
5. Zeros following a decimal point are considered significant. 275 g_ x100 = 55.0%
(trailing zeros) 500 g
6. Trailing zeros to the right of the decimal ARE significant.
7. Trailing zeros in a whole number with the decimal shown
ARE significant. PERCENT SOLUTIONN – volume / volume
540. = 3 • Problem 3.
8. Trailing zeros in a whole number with no decimal shown are What is the percent by volume in a solution that contains 350 Ml
NOT significant. of rubbing alcohol in a 500 mL of solution?
540 = 2 350 mL_ x100 = 70.0%
500 mL

PERCENT SOLUTIONN – weight / volume

• Problem 4.
What is the % w/v of a solution that has 7.5 g of sodium chloride
diluted to 100 mL with deionized water?
7.5 g_ x100 = 7.5%
100 mL

MOLARITY
• PRACTICE: • Molarity, also known as molar concentration, is defined as the
1. 0.004 = 1 1. 0.003 = 1 number of moles of solute present in a given number of litres of
2. 0.500 = 3 2. 0.900 = 3 solution, or moles per litre.
3. 0.20 = 2 3. 0.50 = 2 • Remember that 1 mol of a substance is equal to the gmw (gram
4. 0.00330 = 3 4. 0.00120 = 3 molecular weight) of that substance.
5. 1.237 = 4 5. 2.547 = 4
6. 1110. = 4 6. 6780 = 4 M (mol/L) = n (moles of solute) x
V (volume of solution) IN LITERS
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Cchm321 LEC 1st SEMESTER | overview and lab mathematics
• Problem 5. VOLUME, CONCENTRATION
Calculate the molarity of a sugar solution if 4 liters of the solution
contains 8 moles of sugar? V 1 x C1 = V 2 x C 2
8 mol_ = 2.0 M or 2.0 mol/L
4L • Problem 11.
What volume is needed to make 500 mL of a 0.1 M solution of
• Problem 6. Tris buffer from a solution of 2M Tris buffer?
What is the molarity of a solution containing 5 moles of solute in *Tris amino methane – buffering component to maintain pH
250 milliliters of solution?
5 mol_ = 20.0 M or 20.0 mol/L Identify the known values:
0.25 L Concentration of initial substance (C1) = 2M
Volume of the product (V2) = 500 mL
• Problem 7. Concentration of the product (C2) = 0.1M
Calculate the molarity of a solution prepared by dissolving 23.7
grams of KMnO4 into enough water to make 750 mL of solution. V 1 = V 2 x C2 = 500 mL x 0.1M = 25.0 mL
C1 2M
K = 39.1 g/mol molar mass of KMnO4 = 158.0 g/mol
+ Mn = 54.9 g/mol moles of solute = g of solute___ DILUTION
O = 16.0 g/mol x 4 molecular weight • Represents the ratio of concentrated or stock material to the
= 23.7 g / 158.0 g/mol total final volume of a solution.
= 0.15 mol = 0.2 mol • Consists of the volume or weight of the concentrate plus the
volume of the diluent.
M = 0.2 mol_ = 0.3 M or 0.3 mol/L
0.75 L DILUTION FACTOR
• Ratio of concentrated or stock solution to the total solution
volume.
NORMALITY • The relationship of the dilution factor to concentration is an
• Eq/L or mmol/L inverse one. If DF increases, concentration decreases.
• Often used in chemical titration, reagent classification, and as a • To determine the dilution factor:
unit to express electrolytes like sodium, potassium, and chloride. Simply take the concentration needed and divide by the
• Number of gram equivalent weights per 1 L of solution. stock concentration, leaving it in a reduced fraction form.
• Equivalent weight is equal to the gmw of a substance divided
by its valence. (GMW/Valence) DF = Vf / Vi
• Valence is the number of units that can combine with or replace
1 mole of hydrogen ions for acids and hydroxyl ions for bases DILUTION = volume of solute:volume of solution
and the number of electrons exchanged in oxidation–reduction
reactions. NOTE: DF IS A WHOLE NUMBER, WHILE DILUTION IS
**It is the number of replicate hydrogen ions for acids and EXPRESSED AS A RATIO (DF SERVES AS ITS DENOMINATOR)
hydroxyl ions for bases. DO NOT ADD A DECIMAL.

N = Grams of solute . N = Molarity x Valence • Problem 12.

EW x volume (L) What is the dilution factor if you add a 0.1 mL aliquot of a
specimen to 9.9 mL of diluent?
• VALENCE **aliquot – carefully measured volumes from the stock solution
ACIDS – count the number of Hydrogen ions DF = (0.1 mL + 9.9 mL) = 10.0 mL = 100
BASES – count the number of Hydroxide ions 0.1 mL 0.1 mL
SALTS – multiply the absolute value of the ions
DILUTION = 1/100 = 1:100
ACID BASE SALT
HCl – 1 NaOH – 1 NaCl - Na(+1) x Cl(-1) = 1 • Problem 13.
HNO3 – 1 KOH – 1 CaCl2 - Ca(+2) x Cl(-1) = 2 How would you make a 1:250 dilution in 500 mL?
H2SO4 – 2 NH4OH – 1 FeCl3 - Fe(+3) x Cl(-1) = 3 DF = 250
H3PO4 – 3 Ba(OH)2 – 2 VI = Vf / DF = 500 mL/250 = 2.0 mL

• Problem 8. **Meaning, to make a 1:250 dilution in a graduate cylinder, add

Calculate the equivalent weight of the following: 2.0 mL of solute then 498 mL of solvent to make a solution of
1. H2SO4 – GMW: 98 500 mL.
= 98/2 = 49.0
2. NaCl – GMW: 58.5 TYPES OF DILUTION
= 58.5/1 = 58.5 • Simple Dilution
3. NaOH – GMW: 40 • Serial Dilution
= 40/1 = 40.0 • Doubling Dilution

• Problem 9. SIMPLE DILUTIONS

Calculate the normality of NaOH solution formed by dissolving
0.2 g NaOH to make 250 mL solution. • Problem 14.
____0.2 g____ = 0.02 N A specimen is diluted by combining 3 mL of serum with 21 mL of
(40.0) (0.25 L) Saline. What is the dilution of the serum?

• Problem 10. Solution. Read the problem carefully and calculate the dilution.
What is the normality of 0.1381 M NaOH? Note: Volume units must be the same when calculating dilutions.
0.1381 M x 1 = 0.1 N
3 mL (parts serum) Dilution = 3 mL / 24 mL
+ 21 mL (parts saline) = 1:8
24 mL (total parts)

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Cchm321 LEC 1st SEMESTER | overview and lab mathematics
SERIALS DILUTIONS
• Problem 15.
A serum specimen was successively diluted 1:2, 1:2, and 1:2
with saline. What is the final dilution of the specimen?

Solution. When determining the final dilution of a series of

dilutions use each dilution as a multiplicative factor. The resulting
product is the final dilution.

_1_ x _1_ x _1_ = _1_ or equivalently, 1:8

2 2 2 8

DF will be 8.

• Problem 16.
If a 1/8 dilution of the stock solution is made followed by a 1/6
dilution what is the final dilution.

Solution: When determining the final dilution of a series of

dilutions use each dilution as a multiplicative factor. The resulting
product is the final dilution.

_1_ x _1_ = _1_ or equivalently, 1:48

8 6 48

DF will be 48.

DOUBLING DILUTIONS
• Problem 17.
Doubling dilution 6 times.

1st dilution = 1 /2
2nd dilution = 1 /2 x 1 /2 = 1/4
3rd dilution = 1/4 x 1 /2 = 1/8
4th dilution = 1/8 x 1 /2 = 1/16
5th dilution = 1/16 x 1 /2 - 1/32
6th dilution = 1/32 x 1 /2 = 1/64

This results in a series of dilutions, each a doubling dilution of

the previous one

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