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Blood specimen preparationGlucose analysisLiver function test
Neutral sugar and sialic acid determination( 由吳教授講解 )
Blood sample collection
Heparin
Heparin: Direct thrombin inhibitors
Blood glucose determination
Hormonal regulation• Insulin: Release from -cells islet of Langerhans of pancreas in r
esponse to high blood glucose– Increase uptake of glucose– Increase synthesis of glycolytic enzymes, glucokinase, PFK, & PK– Inhibits gluconeogenesis
• Glucagon: -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
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.
Blood glucose determination
Glucose oxidase
2H2O + Glucose + O2 → gluconic acid + H2O2
2H2O2 + 4-aminoantipyrine (AAP) → 4H2O + Quinoneimine Dye (OD505, pink)
peroxidase
血糖的正常值
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
2003 年下修
Liver function testGOT and GPT determination
肝功能測試 :決定你人生是黑白或彩色
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.
• 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 → Pyruvate + Glutamate
•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.
Transaminase: GOT(AST), GPT(ALT)
Organs GOT GPT
Myocardium
Liver
Kidney
Pancreas
Spleen
Lung
Red blood cell
Serum
156,000
142,000
99,000
28,000
14,000
10,000
300
20
7,100
44,000
4,800
2,000
1,200
720
110
16
Concentration in organs and tissues (Karmen unit/1 gm wet weight)
Chapter 16 TCA cycle
Malate aspartate shuttle
• 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.
測定血液中的 GOT 及 GPT 活性
• Reitman & Frankel 比色法 ( 酮酸 -DNP 呈色法 ) -Keto acid– DNP (2,4-dinitrophenylhydrazine) (金黃色 )– 2,4-dinitrophenylhydrazone (紅棕色 )
R C COOH
O
NH2NH
NO2
NO2+ R C
COOH
N
NO2
NO2NH+
OH-
(Pyruvate or OXA)
Aspartic acid or alanine
Transaminase (GOT or GPT)
GOT 及 GPT 的正常值
Serum GOT(SF Units/ml)
Serum GPT(SF Units/ml)
正常值 0~28 0~21
臨界值 28~40 21~35
S. F. Unit (Sigma-Frankel Unit):在 25°C , pH 7.5 的狀態下,每分鐘能產生 4.82×10-4 mol glutamate 的酵素活性。
I. Unit (International Unit):在特殊狀態下,每分鐘能轉變 1 mol 受質的酵素量。
1 SF U/ml = 0.48 IU/L
GOTGOT 及及 GPTGPT 的比較 的比較 Characteristic GOT GPTPresent in tissues
other than liverMore in heart than in
liver; also in skeletal muscle, kidney, brain
Relative low conc. in other tissue
Location in hepatocyte Mitochondria and cytoplasm
Cytoplasm only
Reference range in adult blood
10-40 IU/liter 5-35 IU/liter
Half-life in blood 12-22 hr 37-57 hr
Change with acute inflammatory damage
Moderately sensitive Extremely sensitive
Change with primary or secondary neoplasm
Substantial rise Moderate or no rise
Change with cirrhosis Moderate rise Mild or moderate rise
Change with myocardial infarct
Substantial rise Mild or moderate rise
(GOT) (GPT)