Serum alkaline phosphatase

Phosphate monoester hydrolase

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alkaline phosphatase (ALP) genus phosphoric acid Monoester hydrolase , is a specific group Phosphatase 。 This enzyme It is widely distributed in human tissues and body fluids, including bone, liver breast , small intestine and kidney, most of which are composed of bone cell Produced, a small part comes from the liver bile Discharges into the intestinal tract. ALP determination methods are mainly colorimetry and Continuous monitoring method 。

Chinese name: Serum alkaline phosphatase
Foreign name: alkaline phosphatase

Classification: Blood biochemical examination and enzyme determination
Identification method: Colorimetry, continuous monitoring
Category: Specific phosphatase

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Discrimination principle

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（1） colorimetry : ALP makes phosphoric acid Disodium benzene is hydrolyzed to produce phosphoric acid and Free phenol , the latter and 4- amino Antipyrine Function, and pass through iron Potassium cyanide Oxidation to red Quinones The color of compounds is directly proportional to the activity of ALP.

（2） Continuous monitoring method : ALP under alkaline condition nitrobenzene Phenol (4- NP P) Releasing phosphoric acid group, AMP participates in phosphoric acid acyl Transfer of, promoting enzyme reaction rate , generating free pairs Nitro phenol (4-NP), and form yellow quinone, which absorbance The increasing rate of enzyme activity Is proportional.

reagent

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colorimetry

①0.1 mol/L Carbonate buffer (pH10.0): weigh anhydrous sodium carbonate 6.36g， sodium bicarbonate 3.36g，4- amino Antipyrine 1.5g, soluble in 800ml distilled water Medium, move into 1000ml Volumetric flask Add distilled water to the scale and store it in a brown bottle.

② 20mmol/L disodium phenyl phosphate solution: weigh Disodium phenyl phosphate (AR) 2.18g, quickly add 500ml of boiled distilled water, and add after cooling chloroform 2ml antiseptic, stored in refrigerator. This is the matrix liquid.

③ Iron Potassium cyanide Solution: weigh Potassium ferricyanide 2.5g， boric acid 17g, respectively dissolved in 400ml distilled water, mixed with water to 1000ml, stored in a brown bottle away from light. appear Turquoise It should be abandoned.

④ Phenol standard storage solution (1mg/ml): weigh 1.0g of redistilled phenol and dissolve it in 0.1mol/L hydrochloric acid And add to the 1000ml scale. Standard solutions can also be purchased.

⑤ Phenol standard application solution (0.05mg/ml): take 5ml of phenol standard storage solution and add distilled water to 100ml.

Continuous monitoring method

①1.8mol/L AMP Buffer (pH10.3): Weigh 2- amino -2 methyl- 1-Propanol (AMP, MW89.14) 160g, add 1mol/L hydrochloric acid 320ml to mix, add 500ml newly boiled and cooled distilled water, adjust the pH to 10.3 ± 0.02 (30 ℃), add the above distilled water to 1000ml, place it in a tightly closed bottle and store it in the refrigerator.

②10.5mmol/L Magnesium chloride Solution: weigh magnesium chloride（ MgCl2·6H2O ， MW 20 3.31) 0.21g, dissolved in water, diluted to 100ml, and stored at room temperature.

③ 31.5mmol/L phosphoric acid nitrobenzene Phenol solution (4-NPP): phosphoric acid P-Nitrophenol Disodium salt (containing 6 molecules of water, MW 371.15）120.8mg， Dissolve in 100ml distilled water, place in a brown bottle and store in refrigerator.

④ Substrate buffer (0.84mol/L AMP, 15mmol/L 4-NPP, 0.5mmol/L MgCl2 , pH10.3) Mix AMP, 4-NPP and MgCl23 solutions at a ratio of 10 ∶ 10 ∶ 1, and preheat them at 37 ℃ for use.^[1]

Normal value

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(1) Enzyme rate method (37 ℃):

Adult: 40~160U/L;

Children:<350U/L;

(2) Disodium phenyl phosphate Law:

Adults: 3~13Kat units;

Children: 5~30kat unit;

(3) Dynamic method: adult: 20~110U/L;

teenagers:

1. Male:<750U/L;

2. Female:<500U/L;

Children:<500U/L;

Infants: 50~240U/L.

(Note: specific reference value Please determine according to each laboratory.)

Clinical significance

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(1) Raise:

① None jaundice ： cirrhosis , hepatocellular carcinoma bile duct Disease（ Cholangiocarcinoma , intrahepatic stones), localized liver damage（ Metastatic liver cancer 、 Hepatic abscess 、 granuloma nature Liver injury ）Other liver diseases（ chronic hepatitis 、 Fatty liver )、 Bone disease ( rickets Osteomalacia , or Osteosarcoma ）, Transferability cancer of bone 、 Hyperthyroidism ， Hyperparathyroidism ， Chronic renal insufficiency ， Malignant tumor (Bone and liver metastasis), etc.

② Jaundice: Viruses Hepatitis Alcoholic hepatitis 、 Drug induced liver damage 、 hepatitis virus Liver damage caused by viruses other than Primary biliary cirrhosis Other intrahepatic bile stasis Obstructive jaundice ( Cholangiocarcinoma 、 Cholangiocarcinoma 、 Carcinoma of pancreatic head , total Biliary calculus ， Sclerosing cholangitis )。

(2) Decrease: milk alkali syndrome Scurvy 、 Hypothyroidism disease 、 vitamin D Excessive intake Pernicious anemia , Severe chronic nephritis 、 Petrification 、 Cachexia Etc.

Reasons for high

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Causes of abnormal metabolism

alkaline phosphatase Mainly used for obstructive jaundice liver cancer 、 bile Examination of stasis hepatitis, obstructive jaundice Primary liver cancer 、 Secondary liver cancer Cholestasis hepatitis will cause high alkaline phosphatase.

Abnormal biliary excretion

alkaline phosphatase It needs to pass through the liver, and then bile Excretion, when there is abnormality in the process of excretion, it will cause high alkaline phosphatase.

Cause of bone tissue

because bone tissue This enzyme is also very active. Therefore, pregnant women fracture Healing period Osteomalacia 。 Rickets bone cell Cancer osteoporosis Isoserum alkaline phosphatase Will rise.

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