Biochemical Testing - Biochemistry for Lab Technicians

Microbiology for Lab Technicians - one year diploma

1. Lab Technician by, Miss Areesha Ahmad Senior Lecturer B.Sc (H), M.Sc , M.Phil and Ph.D scholar (Microbiology) Biochemistry

2. Introduction to Chemistry Analyzer Definition : A chemistry analyzer is an instrument used to measure chemical components in biological samples (e.g., blood, serum, plasma). Purpose : Accurate quantification of biochemical parameters for diagnosis, treatment, and monitoring. Examples : Beckman Coulter, Roche Cobas , Siemens Atellica Serum and plasma both come from the liquid portion of the blood that remains once the cells are removed. Serum is the liquid that remains after the blood has clotted. Plasma is the liquid that remains when clotting is prevented with the addition of an anticoagulant.

3. Types of Chemistry Analyzers Automated Chemistry Analyzers Fully automated, high throughput Semi-Automated Chemistry Analyzers Partially manual, suitable for smaller labs Portable Chemistry Analyzers ( on-site patient testing) Point-of-care testing (POCT), quick results

4. Components of a Chemistry Analyzer Sample Loader : For introducing the specimen Reagent System : Preloaded chemicals for reactions Reaction Chamber : Area for chemical reactions Optical System : For measuring absorbance (Photometry) Data Processor : Displays and stores results

5. Principle of Operation Photometry : Measurement of light absorbance through samples End-Point Reaction : Final absorbance measured after reaction completion Kinetic Reaction : Absorbance measured at multiple time intervals ISE (Ion-Selective Electrode) : Used for electrolyte analysis

6. Routine Chemistry Tests

7. Routine Chemistry Tests Routine chemistry involves the analysis of basic biochemical parameters essential for patient diagnosis and monitoring. Such as; Blood sugar Liver function tests (LFT) Lipid Profile Cardiac Enzymes Renal Function Tests (RFTs) Hormonal test

8. 1. Blood Sugar 1) Fasting Blood Sugar (FBS) : Measures glucose levels in the blood after fasting for 8–12 hours. Used to diagnose diabetes and monitor glucose metabolism. Normal Range: 70–99 mg/ dL (3.9–5.5 mmol /L ) Prediabetes : 100–125 mg/ dL (5.6–6.9 mmol /L ) Diabetes : ≥126 mg/ dL (≥7.0 mmol /L)

9. 1. Blood Sugar 2) Random Blood Sugar (RBS) : Measures glucose levels at any time of the day, regardless of fasting. Helps identify hyperglycemia or hypoglycemia . Normal Range: 70–140 mg/ dL (3.9–7.8 mmol /L ) Diabetes Diagnosis: ≥200 mg/ dL (≥11.1 mmol /L), with symptoms of hyperglycemia or a crisis.

10. 2. Liver Function Tests (LFTs) 1) Alanine Aminotransferase (ALT) : Enzyme found in the liver; elevated levels indicate liver damage or inflammation. Normal Range: 7–56 U/L 2) Aspartate Aminotransferase (AST) : Enzyme present in the liver and other tissues; high levels suggest liver or heart damage. Normal Range: 10–40 U/L 3) Alkaline Phosphatase (ALP) : Enzyme linked to bile ducts; elevated in liver disease or bone disorders. Normal Range: 44–147 U/L

11. 2. Liver Function Tests (LFTs) 4) Bilirubin (Total and Direct) : Measures the levels of bilirubin, a byproduct of red blood cell breakdown. Elevated levels indicate jaundice, liver disease, or bile duct blockage . Total Bilirubin Normal Range: 0.3–1.2 mg/ dL Direct Bilirubin Normal Range: 0.1–0.3 mg/ dL 5) Albumin and Total Protein : Evaluates liver’s synthetic function and nutritional status. Low levels indicate liver dysfunction or malnutrition. Albumin Normal Range: 3.5–5.0 g/ dL Total Protein Normal Range: 6.0–8.3 g/ dL

12. 3. Lipid Profile 1 ) Total Cholesterol : Measures overall cholesterol in the blood. High levels are a risk factor for cardiovascular disease. Normal Range: <200 mg/ dL Borderline High: 200–239 mg/ dL High : ≥240 mg/ dL 2) High-Density Lipoprotein (HDL) : “Good cholesterol” that helps remove bad cholesterol from the blood. Low levels are associated with heart disease. Normal Range : Men : ≥40 mg/ dL Women : ≥50 mg/ dL

13. 3. Lipid Profile 3) Low-Density Lipoprotein (LDL) : “Bad cholesterol” that can accumulate in arteries, leading to atherosclerosis . Normal Range: <100 mg/ dL Near Optimal: 100–129 mg/ dL Borderline High: 130–159 mg/ dL High : 160–189 mg/ dL Very High: ≥190 mg/ dL 4)Triglycerides : Type of fat in the blood. Elevated levels are linked to heart disease and metabolic syndrome. Normal Range: <150 mg/ dL Borderline High: 150–199 mg/ dL High : 200–499 mg/ dL Very High: ≥500 mg/ dL

14. 4. Cardiac Enzymes 1) Creatine Kinase-MB (CK-MB) : Enzyme specific to the heart muscle. Elevated levels indicate myocardial infarction (heart attack). Normal Range: 0–5 ng /mL (varies by lab) 2) Troponin-I or Troponin-T : Proteins released into the blood when heart muscle is damaged. Highly sensitive and specific markers for heart attack. Normal Range:Troponin-I : <0.04 ng /mL Troponin-T : <0.01 ng /mL 3) Lactate Dehydrogenase (LDH) : Enzyme involved in energy production. Increased levels can indicate tissue damage, including myocardial or liver injury. Normal Range: 140–280 U/L

15. 5. Renal Function Tests (RFTs) 1) Urea Definition : Waste product from protein metabolism, excreted by the kidneys. Purpose : Measures kidney function; high levels indicate impaired kidney function or dehydration . Normal Range: 7–20 mg/ dL (2.5–7.1 mmol /L) 2) Creatinine Definition : Byproduct of muscle metabolism comes from the digestion of protein in your food filtered by the kidneys. Purpose : Evaluates kidney efficiency; elevated levels suggest kidney dysfunction . Normal Range: Men : 0.6–1.2 mg/ dL (53–106 µ mol /L ) Women : 0.5–1.1 mg/ dL (44–97 µ mol /L)

16. 5. Renal Function Tests (RFTs) 3) Electrolytes Sodium (Na⁺) : Maintains fluid and electrolyte balance; abnormal levels indicate dehydration or hormonal imbalances. Normal Range: 135–145 mEq /L Potassium (K⁺) : Regulates heart and muscle function; imbalances can cause serious cardiac issues. Normal Range: 3.5–5.0 mEq /L Chloride ( Cl ⁻) : Maintains acid-base balance; abnormal levels suggest metabolic disorders. Normal Range: 98–106 mEq /L

17. 5. Renal Function Tests (RFTs) 4) Blood Urea Nitrogen (BUN) Definition : Measures urea nitrogen in the blood, a byproduct of protein metabolism. Purpose : Assesses kidney function; high levels indicate kidney issues or high protein intake . Normal Range: Adults: 7–20 mg/ dL Children: 5–18 mg/ dL Older Adults: Slightly higher values may be normal due to decreased kidney function with age .

18. 6. Hormonal test Serum Prolactin Definition : Hormone produced by the pituitary gland ( located at the base of your brain), primarily responsible for lactation . It also plays a role in reproductive health for both males and females. Purpose : Evaluates pituitary function. High levels ( hyperprolactinemia ) indicate pituitary tumors, hypothyroidism, or stress. Normal Range: Men: 2–18 ng /mL Non-pregnant Women: 2–25 ng /mL Pregnant Women: 10–209 ng /mL

19. 6. Hormonal test 2. Follicle-Stimulating Hormone (FSH) Definition : Hormone produced by the pituitary gland, essential for reproductive processes. Purpose : In women: Regulates ovarian follicle development and estrogen production. In men: Stimulates sperm production. Abnormal levels indicate infertility, menopause, or pituitary disorders. Normal Range : Women : Follicular phase: 3–10 mIU /mL Mid-cycle (ovulation): 4–25 mIU /mL Luteal phase: 1.5–9 mIU /mL Postmenopausal : 40–250 mIU /mL Men : 1–10 mIU /mL Children (prepubescent): <4 mIU /mL

20. 6. Hormonal test 3. Luteinizing Hormone (LH) Definition : Pituitary hormone that triggers ovulation in females and testosterone production in males. Purpose : In women: Assesses ovulatory function and menopause. In men: Evaluates testosterone production and testicular function. Abnormal levels suggest infertility or pituitary dysfunction. Normal Range: Women : Follicular phase: 1.9–12.5 IU/L Mid-cycle (ovulation): 8.7–76.3 IU/L Luteal phase: 0.5–16.9 IU/L Postmenopausal: 15.9–54.0 IU/L Men: 1.5–9.3 IU/L Children: 0.1–6.0 IU/L

21. 6. Hormonal test 4. Progesterone Definition : Hormone produced by the ovaries (and placenta during pregnancy), crucial for maintaining pregnancy. Purpose : Evaluates ovulation and luteal phase ( second part of your menstrual cycle), the follicle that burst and released the egg (during ovulation) in women. Monitors early pregnancy. Low levels may indicate luteal phase defects or risk of miscarriage. Normal Range : Women (Non-pregnant): Follicular phase: <1 ng /mL Luteal phase: 2–25 ng /mL Postmenopausal: <1 ng /mL Pregnant Women: First trimester: 11–44 ng /mL Second trimester: 25–83 ng /mL Third trimester: 58–214 ng /mL Men: <1 ng /mL

22. Special Chemistry

23. Special Chemistry Special chemistry involves advanced biochemical testing that is not routinely performed, often requiring specialized equipment or methods. Such as, HBA1C Serum Amylase Serum, insulin T hyroid profile Serum iron S erum ferritin Total Iron Binding Capacity (T.I.B.C) Glucose Challenge Test (GCT) Glucose Tolerance Test (GTT) Alpha-Fetoprotein (AFP)

24. Special Chemistry Tests HbA1c ( Glycated Hemoglobin) Definition : Reflects average blood glucose levels over the past 2–3 months. Purpose : Monitors long-term glucose control in diabetic patients. Normal Range: Normal : <5.7% Prediabetes : 5.7%–6.4% Diabetes: ≥6.5 % 2. Serum Amylase Definition : Enzyme secreted by the pancreas and salivary glands. Purpose : Elevated levels help diagnose pancreatitis and other pancreatic disorders. Normal Range: Adults : 30–110 U/L Children: Slightly higher levels may be normal.

25. Special Chemistry Tests 3. Serum Insulin Definition : Hormone secreted by the pancreas that regulates blood sugar. Purpose : Used to diagnose insulin resistance, type 2 diabetes, or insulinoma (tumor of insulin-producing cells). Normal Range: Fasting Insulin: 2–25 µIU/mL Postprandial Insulin (after a meal): 5–50 µIU/mL 4. Thyroid Profile Definition : Includes T3 ( Triiodothyronine ), T4 ( Thyroxine ), and TSH (Thyroid Stimulating Hormone). Purpose : Assesses thyroid function to diagnose hypothyroidism or hyperthyroidism. Normal Range: T3: 2.3–4.1 pg / mL. T4: 0.8–2.0 ng / dL . TSH: 0.4–4.0 mIU /L

26. Special Chemistry Tests 5. Serum Iron Definition : Measures the amount of iron in the blood. Purpose : Identifies iron deficiency or overload conditions, such as anemia or hemochromatosis. Normal Range: Men : 65–175 µg/ dL Women: 50–170 µg/ dL Children: 50–120 µg/d 6. Serum Ferritin Definition : Protein that stores iron and releases it when needed. Purpose : Low levels indicate iron deficiency anemia; high levels suggest inflammation or iron overload. Normal Range: Men : 30–400 ng /mL Women: 20–150 ng /mL Children: 7–140 ng /mL

27. Special Chemistry Tests 7. Total Iron Binding Capacity (TIBC) Definition : Measures the blood's ability to bind iron with transferrin. Purpose : Helps differentiate between types of anemia. Elevated in iron deficiency anemia, low in chronic diseases. Normal Range: Adults : 240–450 µg/ dL Children: 250–450 µg/ dL 8. Glucose Challenge Test (GCT) Definition : Measures the body’s response to a glucose load (50g, non-fasting). Purpose : Screening test for gestational diabetes during pregnancy. Normal Range: One hour after glucose drink: Less than 140 mg/ dL (7.8 mmol /L)

28. Special Chemistry Tests 9. Glucose Tolerance Test (GTT) Definition : Assesses how the body processes glucose over time after consuming a glucose solution. Purpose : Diagnoses diabetes or gestational diabetes. Normal Ranges: Fasting (before drinking glucose): Normal: 70–99 mg/ dL (3.9–5.5 mmol /L) Impaired Fasting Glucose: 100–125 mg/ dL (5.6–6.9 mmol /L) Diabetes: ≥126 mg/ dL (≥7.0 mmol /L) 1 Hour after drinking glucose: Normal: Less than 180 mg/ dL (10.0 mmol /L) 2 Hours after drinking glucose: Normal: Less than 140 mg/ dL (7.8 mmol /L) Impaired Glucose Tolerance (Pre-diabetes): 140–199 mg/ dL (7.8–11.0 mmol /L) Diabetes: ≥200 mg/ dL (≥11.1 mmol /L) 3 Hours after drinking glucose (if a 3-hour GTT is performed): Normal: Less than 140 mg/ dL (7.8 mmol /L )

29. Special Chemistry Tests 10. Alpha-Fetoprotein (AFP ) Definition : Protein produced by the fetal liver, found in small amounts in adults. Purpose : In pregnancy: Screens for fetal abnormalities like neural tube defects. In adults: Marker for liver cancer or germ cell ( C ells that create reproductive cells called gametes) tumors . Normal Range: Adults: <10 ng /mL Pregnant Women: First trimester (weeks 1-12): 20–120 ng /mL Second trimester (weeks 13-20): 20–100 ng /mL Third trimester (weeks 21-40): 20–80 ng /mL

30. Differences b/w Special and Routine Chemistry tests

31. Serum and Plasma difference Serum is the liquid that remains liquid after the clotting of blood. Whereas, plasma is the liquid that remains when anticoagulant is added to prevent clotting.

32. Importance of Routine and Special Chemistry Helps in early diagnosis of diseases Monitors treatment and disease progression Provides critical information for clinical decision-making

33. Challenges in Chemistry Analysis Sample Handling : Contamination or hemolysis Calibration Issues : Inaccurate results due to improper calibration Reagent Quality : Stability and reliability of reagents Automation Errors : Instrument malfunction or human error

34. Case Study/Example Case Example : Patient with suspected diabetes Tests Performed : Random Blood Sugar, HbA1c Analyzer Used : Roche Cobas Result : Diagnosis of diabetes