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Blood sample collection

Blood specimen preparation Glucose analysis Liver function test Neutral sugar and sialic acid determination ( 由 吳明道 教授講解 ). Blood sample collection. Heparin. Heparin: D irect thrombin inhibitors. Blood glucose determination. Hormonal regulation.

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Blood sample collection

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  1. Blood specimen preparationGlucose analysisLiver function testNeutral sugar and sialic acid determination(由吳明道教授講解)

  2. Blood sample collection

  3. Heparin Heparin: Direct thrombin inhibitors

  4. Blood glucose determination

  5. Hormonal regulation • Insulin: Release from b-cells islet of Langerhans of pancreas in response to high blood glucose • Increase uptake of glucose • Increase synthesis of glycolytic enzymes, glucokinase, PFK, & PK • Inhibits gluconeogenesis • Glucagon:a-cells islets of Langerhans of pancreas in response to low blood glucose • Stimulate liver (not muscle) to release glucose • Involved in long term glucose maintenance • Epinephrine (adrenaline): released by adrenal glands • Active on liver and muscle • Stimulates glycogen breakdown and glycolysis

  6. The typical human adult uses 160 g of glucose per day, with 120 g being used by the brain. The available reserve of glucose is adequate for about one day (about 20 g of circulating glucose and 190 g of glycogen). After the reserve becomes depleted, how would a starved body obtain more glucose? In animals, many precursors lead to the synthesis of glucose (gluconeogenesis). In humans, the principal precursors are glycerol from triacylglycerols and glucogenic amino acids from proteins.

  7. Blood glucose determination 2H2O + Glucose + O2 → gluconic acid + H2O2 2H2O2 + 4-aminoantipyrine (AAP) → 4H2O + Quinoneimine Dye (OD505, pink) Glucose oxidase peroxidase

  8. 血糖的正常值 2003年下修 正常人空腹時的血糖值為 70-110 mg/dl 將空腹血糖異常值從110 mg/dL降到了100 mg/dL，可能使前驅前糖尿病的診断增加约20% 預料國內將多出近20萬名代謝症候群病患

  9. Liver function testGOT and GPT determination 肝功能測試:決定你人生是黑白或彩色

  10. Transaminase:GOT(AST), GPT(ALT) • Having the function of transferring amino group of amino acids from alpha-amino acids to alpha-keto acids, therefore, named transaminase.

  11. GOT(Glutamyl oxaloacetic transaminase, Aspartate aminotransferase, AST) • L-Aspartic Acid + Ketoglutaric Acid → Oxaloacetate + L-Glutamate • GPT (Glutamyl pyruvic transaminase, Alanine aminotransferase, ALT) • L-Alanine + Ketoglutaric acid→ Pyruvate + Glutamate

  12. Transaminase: GOT(AST), GPT(ALT) • One of the thousands kinds of liver enzymes, and a kind of transferase. • Storage mostly in liver etc. • Low serum concentration. • Large amount of transaminase is released into blood mostly on liver cell damages. -Thus, detection of serum level tells the existence of liver cell damage.

  13. Concentration in organs and tissues (Karmen unit/1 gm wet weight)

  14. Chapter 16 TCA cycle

  15. Malate aspartate shuttle

  16. METHOD PRINCIPLE • In this method ALT catalyzes the reaction of L-alanine and - ketoglutaric acid to form pyruvate and glutamate under controlled conditions (37 C) and pH 7.4 + 0.05. The addition of acidic 2,4-dinitrophenylhydrazine stops the reaction and forms the 2,4-dinitrophenylhydrazone. So that it may be measured at 505 + 5 nm.

  17. 測定血液中的GOT及GPT活性 • Reitman & Frankel 比色法 (酮酸-DNP呈色法) • a-Keto acid • DNP (2,4-dinitrophenylhydrazine) (金黃色) • 2,4-dinitrophenylhydrazone (紅棕色) NO2 NO2 O COOH H+ R C COOH ＋ NH2NH NO2 R C N N NO2 (Pyruvate or OXA) OH- Transaminase (GOT or GPT) Aspartic acid or alanine

  18. GOT及GPT的正常值 S. F. Unit (Sigma-Frankel Unit):在25°C，pH 7.5的狀態下，每分鐘能產生4.82×10-4mmol glutamate的酵素活性。 I. Unit (International Unit):在特殊狀態下，每分鐘能轉變1 mmol受質的酵素量。 1 SF U/ml = 0.48 IU/L

  19. GOT及GPT的比較

  20. Thank you

  21. (GOT) (GPT)

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