Introduction to Biochemistry

Biochemistry is the branch of science that explores the chemical

processes within and related to living organisms. It is a laboratory-based
science that combines biology and chemistry. By using chemical
knowledge and techniques, biochemists can understand and solve
biological problems.

Biochemistry focuses on:

 The structure and function of biomolecules like proteins,

carbohydrates, lipids, nucleic acids, and enzymes.

 Cellular processes such as metabolism, signal transduction, and

energy production.

 The molecular basis of diseases, which helps in developing

treatments and medicines.

Importance of Biochemistry:

 Helps in understanding how cells and organisms function.

 Crucial for medical, agricultural, and nutritional advancements.

 Forms the basis for modern biotechnology and genetic engineering.

Biochemistry is used in a wide range of fields, including medicine,

pharmacology, agriculture, and environmental science.

NAME OF THE INSTRUMENT

1.ATELLICA (SIEMENS HEALTHINERS)

 NAME OF THE TEST
The Atellica® instrument by Siemens Healthineers is an advanced,
automated laboratory analyzer primarily used in clinical biochemistry and
immunoassay testing. It is part of the Atellica Solution, a modular and
flexible system for high-throughput diagnostics.

Common Tests Performed Using the Atellica Instrument:

1. Biochemistry Tests (Atellica CH Analyzer):

 Glucose (GLU)

 Urea / Blood Urea Nitrogen (BUN)

 Creatinine (CRE)

 Cholesterol (CHOL)

 Triglycerides (TRIG)

 High-Density Lipoprotein (HDL)

 Low-Density Lipoprotein (LDL)

 Aspartate Aminotransferase (AST)

 Alanine Aminotransferase (ALT)

 Alkaline Phosphatase (ALP)

  Total Bilirubin

 Total Protein

 Albumin

 Electrolytes (Sodium, Potassium, Chloride)

2. Immunoassay Tests (Atellica IM Analyzer):

 Thyroid Panel (TSH, Free T3, Free T4)

 Vitamin D

 Ferritin

 Prostate-Specific Antigen (PSA)

 Beta-hCG (Pregnancy test)

 Insulin

 Cortisol

 Hepatitis Markers (HBsAg, Anti-HCV, etc.)

 HIV Antigen/Antibody Test

 Tumor Markers (CEA, CA 125, AFP, etc.)

3. Specialized Tests:

 Cardiac Markers (Troponin I, CK-MB, BNP)

 Diabetes Monitoring (HbA1c)

 Infectious Disease Markers

 Hormone Tests (LH, FSH, Estradiol, Progesterone, etc.)

The Atellica® Solution by Siemens Healthineers is an advanced,

integrated platform designed for clinical laboratories, combining
immunoassay and clinical chemistry analyzers with innovative automation
and intelligent software.

Principle of Atellica Biochemistry Analyzer

The Atellica CH Analyzer operates on automated photometric,
turbidimetric, and ion-selective electrode (ISE) methods to analyze
biochemical components in clinical samples like serum, plasma, or urine.

Key Principles:

1. Photometric Analysis

o Based on Beer-Lambert Law: Absorbance of light by a

colored solution is proportional to the concentration of the
analyte.

o Used for: Glucose, urea, creatinine, liver enzymes,

cholesterol, etc.

2. Turbidimetric Method

o Measures reduction in light due to particles (e.g., antigen-

antibody complexes) in the sample.

o Used for: CRP, proteins, lipoproteins, etc.

3. Ion-Selective Electrode (ISE) Method

o Measures the activity of specific ions via electrodes.

o Used for: Electrolytes like sodium (Na⁺), potassium (K⁺),

chloride (Cl⁻).

4. Automation and Sample Handling

o Uses a robotic arm and magnetic transport (Magline®)

for precise sample movement.

o Automates sample identification, mixing, reaction,

measurement, and data reporting.

Clinical Significance

The Atellica Biochemistry Analyzer plays a vital role in clinical diagnostics

due to its speed, accuracy, and broad test menu:

Clinical Relevance:

 Rapid Diagnosis: Delivers results quickly for time-critical tests

(e.g., cardiac enzymes, glucose).

 High Throughput: Capable of performing up to 1800 tests/hour,

ideal for large hospitals or diagnostic labs.
  Wide Range of Tests: Supports assays for liver function, renal
profile, lipid profile, thyroid panel, diabetes monitoring, and more.

 Standardization: Ensures reproducible, comparable results across

labs within a hospital network.

 Error Reduction: Full automation minimizes manual handling,

improving result reliability and biosafety.

Common Tests Performed:

 Metabolic Panel: Glucose, urea, creatinine, uric acid

 Liver Panel: ALT, AST, ALP, bilirubin

 Renal Function: BUN, creatinine

 Cardiac Markers: CK-MB, LDH

 Electrolytes: Na⁺, K⁺, Cl⁻

Method to Use the Fully Automated Atellica

Biochemistry Analyzer (Atellica® CH Analyzer)
Here’s a step-by-step standard operating procedure (SOP)-style
overview for using the Atellica biochemistry analyzer in a
clinical lab setting:

1. System Preparation
 Turn on the system and ensure that the Atellica Solution
software is loaded properly.
 Check reagent levels, water supply, and waste containers.
 Confirm that calibrators and controls are available and
properly stored.

2. Sample Loading
  Label patient samples with barcodes (if available).
 Load sample tubes into the Atellica Sample Handler (ASH).
 The system automatically scans barcodes and assigns
tests based on the LIS (Lab Information System) or manual
entry.

3. Test Request and Selection

 Select tests through the Atellica user interface or they are
automatically pulled from the LIS.
 Tests may include:
o Glucose, Urea, Creatinine, ALT, AST, Electrolytes (Na⁺,
K⁺, Cl⁻), etc.

4. Running the Tests

 The system automatically:
o Aspirates samples.
o Mixes with reagents.
o Measures using photometric, turbidimetric, or ISE
methods.
 Real-time monitoring of each process is available on the
software screen.

5. Quality Control
 Run internal QC samples daily (or as per lab policy).
 Verify QC results fall within acceptable limits before
releasing patient results.

6. Result Validation
  Results are automatically calculated and displayed.
 Pathologist/technician reviews flags or critical values.
 Once verified, results are either:
o Printed, or
o Uploaded to the LIS/HIS for patient reporting.

7. Maintenance (Daily/Weekly)
 Daily tasks:
o Clean sample/reagent probes.
o Check and refill system fluids.
 Weekly/Monthly tasks:
o Perform deeper cleaning.
o Update calibration.
o Check photometer/ISE calibration performance.

Troubleshooting
 The system provides alerts or error codes for clots, sample
errors, reagent shortage, etc.
 Follow on-screen prompts or refer to the operator manual
for resolution.

Shutdown (if needed)

 If shutting down:
o Perform a cleaning cycle.
o Turn off software, then power down hardware.
o Dispose of waste as per biosafety protocol.
2.SEMI AUTOMATED BIOCHEMISTRY ANALYZER

Name of Common Tests Done on a Semi-

Automated Biochemistry Analyzer
1. Glucose
2. Urea
3. Creatinine
4. Total Cholesterol
5. Triglycerides
6. SGOT/AST & SGPT/ALT
7. Alkaline Phosphatase (ALP)
8. Total Protein & Albumin
9. Bilirubin (Total & Direct)
10.Uric Acid

Principle of a Semi-Automated Biochemistry

Analyzer
 Photometric (Colorimetric) Principle
 o Based on Beer-Lambert’s Law, which states that the
concentration of a substance is directly proportional
to the absorbance of light at a specific wavelength.
o The analyzer uses light filters to select the
wavelength and measures the absorbance of the
colored complex formed during the biochemical
reaction.
 Manual Sample Loading: The technician adds the sample
and reagents to cuvettes.
 Automated Reading: The machine reads the absorbance
and calculates the result based on programmed
calibration.

Clinical Significance
 Early Disease Detection: Helps identify metabolic, liver,
kidney, and cardiac disorders.
 Affordable and Accessible: Ideal for smaller laboratories
and resource-limited settings.
 Flexible Use: Can perform a wide variety of tests without
the need for full automation.
 Educational Use: Widely used in teaching labs for student
training in clinical biochemistry.

Summary:
Aspect Semi-Automated Analyzer
Tests Glucose, Urea, Cholesterol, Enzymes, etc.
Principle Colorimetry using Beer-Lambert’s Law
Clinical Basic diagnostics for liver, kidney, and metabolic
Use health
Advantag Cost-effective, simple, suitable for low- to mid-
Aspect Semi-Automated Analyzer
es volume labs

Method to Use a Semi-Automated Biochemistry

Analyzer
1. Preparation
 Turn on the analyzer and allow it to warm up.
 Select the required test program from the instrument
menu (e.g., glucose, urea).
 Prepare all reagents and calibrators as per kit instructions.
 Calibrate the instrument using a blank (usually distilled
water or reagent blank).

2. Sample and Reagent Handling

 Pipette the required amount of reagent into a cuvette or
test tube.
 Add the sample (serum or plasma, usually 10–20 µL) into
the cuvette.
 Mix gently and incubate (if required) at room temperature
or using an external incubator.

3. Measurement
 Place the cuvette into the reading chamber of the
analyzer.
 Select the appropriate wavelength (e.g., 505 nm for
glucose).
 The analyzer measures absorbance and displays the
result.
  For unknown samples, it calculates the concentration
using the standard or factor method.

4. Recording Results
 Record or print the result manually or using the built-in
printer (if available).
 Repeat the process for each sample/test.

5. Post-Analysis
 Clean the cuvette and optical chamber with distilled water
or cleaning solution.
 Turn off the analyzer and store reagents properly.

Notes:
 Always run quality control samples daily.
 Use gloves and PPE to avoid contamination or biohazards.
 Ensure accurate pipetting and proper reagent handling for
reliable results.

3.CENTRIFUGE
 Principle of Centrifuge in Biochemistry
The centrifuge is a crucial laboratory instrument used to
separate components of a sample based on their density by
subjecting it to a high-speed rotational force. This method relies
on the principle of centrifugal force, which helps in the
separation of various cellular or biochemical components.

Principle:
 Centrifugal Force: When a sample is spun rapidly, heavier
particles are forced toward the bottom of the tube (or the
outer edge of the rotor) due to centrifugal force, while
lighter components move toward the top.
 The separation occurs because different components (e.g.,
plasma, red blood cells, proteins) have different densities.
 The separation process occurs in various phases:
o Sedimentation: Denser particles (e.g., red blood cells)
settle at the bottom.
o Supernatant: The liquid (e.g., plasma or serum)
remains above the settled particles.

Applications in Biochemistry:
 1. Cell Fractionation: Separates cell components like nuclei,
mitochondria, and cytoplasm for analysis.
2. Plasma Separation: Separates plasma from blood cells by
spinning blood samples.
3. Protein Purification: Helps in the separation of proteins
based on their size or density.
4. Urine and Serum Analysis: Used to concentrate analytes
for further biochemical testing.

Types of Centrifuges in Biochemistry:

 Low-Speed Centrifuges: Used for plasma/serum
separation, and general cell separation.
 High-Speed Centrifuges: Used for isolating organelles,
proteins, or nucleic acids.
 Ultracentrifuges: For very fine separation, like in protein
studies or virus particle isolation.

4.BIOCHEMISTRY URINE ANALYZER

Name of Tests in Biochemistry Urine Analyzer:

1. Urine Glucose
2. Urine Protein
 3. Urine Urobilinogen
4. Urine Bilirubin
5. Urine pH
6. Urine Specific Gravity
7. Urine Ketones
8. Urine Blood/Hemoglobin
9. Urine Nitrite
10. Urine Leukocytes
11. Urine Microalbumin

Principle of Urine Analyzer

The biochemistry urine analyzer uses various chemical and
physical methods to analyze urine samples for the presence of
specific substances and to measure parameters that help
assess kidney function, metabolism, and overall health.
Key Principles:
1. Colorimetric Reactions: Reactions between urine analytes
and specific reagents cause a color change, which is
measured by the analyzer to quantify the analyte.
2. Reflectance Photometry: The analyzer measures the
intensity of light reflected from the test strip or urine
sample to determine the concentration of substances like
glucose, proteins, and ketones.
3. Ion-selective Electrodes (ISE): Used for measuring
electrolytes like sodium, potassium, and chloride in urine.
4. Enzymatic Reactions: Some tests use enzyme-substrate
interactions to identify analytes like glucose and ketones.

Clinical Significance of Urine Analyzer

Urine analysis is an essential diagnostic tool for detecting and
monitoring various health conditions, including:
1. Kidney Function Assessment: Tests for protein, creatinine,
and microalbumin help assess kidney health, identifying
early signs of renal disease.
2. Diabetes Monitoring: Urine glucose and ketones provide
crucial information about blood sugar control and ketosis
in diabetic patients.
3. Urinary Tract Infections (UTIs): Leukocytes and nitrites are
tested to identify signs of infection.
4. Liver Disorders: Bilirubin and urobilinogen levels can
indicate liver dysfunction or hemolysis.
5. Hydration Status: Urine specific gravity helps assess
hydration levels and kidney concentration ability.
6. Acid-Base Balance: Urine pH can indicate metabolic
acidosis or alkalosis.
7. Monitoring Metabolic Disorders: Tests for ketones and
protein help monitor metabolic states like diabetic
ketoacidosis and proteinuria.

Method to Use a Biochemistry Urine Analyzer

Here is a step-by-step procedure to use a biochemistry urine
analyzer:

1. Preparation
 Turn on the Urine Analyzer and allow it to calibrate (if
necessary).
 Prepare test strips or urine sample according to the
analyzer's instructions.
 Ensure necessary reagents are available and ready for
use.
  Label sample containers with patient details (if not using a
barcode system).

2. Sample Collection
 Collect a midstream urine sample in a clean, dry container
to minimize contamination.
 Mix the sample gently (if sediment is present) to ensure
uniformity.

3. Loading the Sample

 Dip a reagent test strip into the urine sample, ensuring it’s
submerged for the time specified by the manufacturer
(typically 1-2 seconds).
 Remove the strip and blot the excess urine on a clean
tissue to avoid contamination or interference.
 Insert the test strip into the analyzer's sample chamber or
reagent disc as per the model's design.

4. Running the Test

 Start the test using the analyzer’s interface.
 The analyzer will automatically measure parameters such
as glucose, protein, pH, specific gravity, etc., based on
colorimetric or photometric methods.
 The results will be processed, and the analyzer will provide
quantitative values or qualitative results
(positive/negative).

5. Reading and Recording Results

 Once the analysis is complete, read the results displayed
on the screen.
  The analyzer will often automatically generate printouts or
digital reports.
 Review results for abnormal findings, such as elevated
glucose, protein, or signs of infection.

6. Post-Analysis Procedures
 Dispose of test strips and clean the analyzer's components
as required.
 Ensure proper waste disposal in line with safety protocols.
 Store samples or reagents for further analysis, if
necessary.

7. Quality Control
 Run quality control tests periodically (e.g., with known
standards or controls) to ensure the accuracy of the
results.
 Ensure regular calibration as instructed by the
manufacturer.

Additional Notes:
 Safety: Always use gloves and appropriate personal
protective equipment (PPE) when handling urine samples.
 Sample Handling: Ensure that urine is processed quickly to
prevent degradation or loss of analytes.

Advantages of Biochemistry:
1. Understanding Biological Processes:
o Biochemistry helps in understanding cellular and
molecular processes, offering insights into how life
functions at a molecular level.
2. Disease Diagnosis:
o It plays a critical role in diagnosing metabolic
disorders, infections, and genetic conditions by
analyzing biomarkers in blood, urine, or tissues.
3. Drug Development:
o Pharmacology and biochemistry are key to
developing drugs and vaccines, enhancing the
understanding of how drugs interact with the body.
4. Nutritional Science:
o Biochemistry helps in understanding nutrition,
identifying how nutrients affect the body, and
contributing to dietary recommendations.
5. Forensic Applications:
o It's used in forensic science for analyzing bodily fluids
(blood, urine, etc.), identifying toxins, and
establishing cause of death.

Disadvantages of Biochemistry:
1. Complexity of Biological Systems:
o Biological systems are complex, and biochemistry
often faces challenges in fully understanding these
processes, as many reactions are interdependent.
2. High Cost of Equipment:
o Advanced biochemical instruments like mass
spectrometers, NMR machines, and chromatography
systems are expensive and may not be available in
smaller labs.
3. Need for Skilled Technicians:
o Biochemical analysis requires highly skilled
professionals to interpret results, as even small errors
in handling or measurement can affect the outcome.
4. Ethical Concerns:
o Some biochemical research, especially involving
genetic testing and gene therapy, raises ethical
issues regarding privacy, consent, and manipulation
of genetic material.
5. Time-Consuming:
o Certain biochemical techniques, like protein
purification or enzyme assays, can be time-
consuming, requiring significant preparation and
analysis time.