Blood sample collection

Blood specimen preparationGlucose analysisLiver function test

Neutral sugar and sialic acid determination( 由吳明道教授講解 )

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 acid→ 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

Thank you

(GOT) (GPT)