2. Principle: The sample is extracted with trichloroacetic acid solution - acetonitrile, purified by cation exchange solid phase extraction column, and determined by high performance liquid chromatography, and quantified by external standard method.
3. Reagents and materials: Unless otherwise stated, all reagents were of analytical grade and the water was the first grade water specified in GB/T 6682 .
3.1 methanol: chromatographically pure;
3.2 acetonitrile: chromatographically pure;
3.3 ammonia water: the content is 25 % ~ 28 %;
3.4 trichloroacetic acid;
3.5 citric acid.
3.6 sodium octane sulfonate: chromatographically pure;
3.7 Aqueous methanol solution: accurately measure 50 mL of methanol and 50 mL of water, mix and reserve;
3.8 Trichloroacetic acid solution ( 1 %): Accurately weigh 10 g of trichloroacetic acid in a 1 L volumetric flask, dissolve in water and dilute to volume, mix and set aside;
3.9 ammoniated methanol solution ( 5 %): accurately measure 5 mL of ammonia water and 95 mL of methanol, mix and reserve;
3.10 ion pair reagent buffer: Accurately weigh 2.10 g of citric acid and 2.16 g of sodium octane sulfonate, add about 980 mL of water to dissolve, adjust the pH to 3.0 , and dilute to 1 L for use .
3.11 melamine standard: CAS 108-78-01 , the purity is greater than 99.0% ;
3.12 melamine standard stock solution: Accurately weigh 100 mg (accurate to 0.1 mg ) melamine standard in a 100 mL volumetric flask, dissolve it with methanol aqueous solution ( 3.7 ) and dilute to volume to prepare a standard of 1 mg/mL . Store stock at 4 °C protected from light.
3.13 Cation Exchange SPE: Mixed cation-exchange solid phase extraction column, benzenesulfonic matrix is polystyrene - divinyl benzene polymer, 60 mg, 3 mL, or equivalent person.
3.14 qualitative filter paper.
3.15 microporous membrane: 0.2 μ m , organic phase.
3.16 Nitrogen: purity greater than or equal to 99.999 %
4. Instruments and equipment
4.1 High Performance Liquid Chromatography ( HPLC ): equipped with a UV detector or a diode array detector.
4.2 Analytical balance: The sensitivities are 0.00001 g and 0.01 g .
4.3 Centrifuge: The speed is not less than 10000 r/min .
4.4 Tianjin Hengao ultrasonic extractor. HS, HU series
4.5 Tianjin Hengao solid phase extraction device. HSE-12D
4.6 Tianjin Heng Ao Nitrogen Blowing Instrument. HGC, HSC series
4.7 Tianjin Hengao vortex oscillator. HMS-350
4.8 Tianjin Hengao vacuum pump. HPD-25
4.9 Tianjin Hengao precision gas steady flow regulating valve.
4.10 stoppered plastic centrifuge tube: 50 mL .
5. Sample processing
5.1 extraction
Weigh 2 g (accurate to 0.01 g ) sample in a 50 mL stoppered plastic centrifuge tube, add 15 mL of trichloroacetic acid solution ( 3.8 ) and 5 mL of acetonitrile. Ultrasonic extraction for 10 min , and then shaking for 10 min , centrifuged at not less than 10000 r/min for 30 min . After the supernatant was filtered through a filter paper moistened with trichloroacetic acid solution, the volume was adjusted to 25 mL with a trichloroacetic acid solution , 5 mL of the filtrate was removed , and 5 mL of water was added to mix and then to be purified.
Note: If the fat content of the sample is high, it can be degreased with the hexane liquid - liquid saturated with trichloroacetic acid solution and then purified by SPE column.
5.2 Activation
A cation exchange solid phase extraction column was activated ( 3,13 ) with 3 mL of methanol and 5 mL of Antialcoholic Liver Extract
1.Hawthorn extract
Hawthorn can enhance myocardial contractility, increase cardiac output, slow down heart rate, dilate coronary blood vessels, and has the effect of lowering blood pressure and blood fat.
2.Rhubarb extract
In addition to laxative and laxative effect, rhubarb also has the effect of lowering blood pressure and cholesterol.
3, wolfberry extract
Lycium barbarum can lower blood sugar, inhibit fat deposition in liver cells, promote liver cell newborn, and have antihypertensive effect.
4, Cordyceps extract
Cordyceps can improve the immune function of the body, strengthen the ability to remove viruses, protect liver cells and indirectly inhibit the occurrence and development of liver fibrosis.
5. Salvia miltiorrhiza extract
Salvia miltiorrhiza is used in more research on anti-hepatic fibrosis treatment. It can protect hepatocytes, reduce the stimulation of stellate cells by hepatocyte injury, necrosis, etc., and inhibit the initiation and development of hepatic fibrosis; the collagen content in the liver is significantly reduced, and the urinary HPP discharge is elevated, suggesting that the drug can promote the degradation of collagen that has been formed, and the proliferating collagen is reabsorbed by hydrolysis.
Antialcoholic Liver Extract,Anti-Alcoholic Liver Extract,Liver Protection,Enhance Liver Nutrition
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