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A biological detection method for quality evaluation and quality control of heat-clearing and detoxifying traditional Chinese medicines

A heat-clearing, detoxifying, and quality-evaluating technology, applied in biochemical equipment and methods, microbial determination/inspection, measuring devices, etc., can solve the problems of non-medicinal ingredients and low bioavailability, and achieve high-quality and effective results

Active Publication Date: 2019-11-19
GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the quality standard of Grass Cortex is to control its quality through a single index of isopyridine, but isopyridine is obviously not its main active ingredient; The content determination of several isoquinoline alkaloids such as palmatine is used to control their quality, but the general oral bioavailability of Coptidis alkaloids is extremely low. Especially good at clearing the fire of the Shangjiao", the medicinal properties of "Apparently cannot be explained clearly by these few alkaloids; the research and development of a quality control system that meets the needs of clinical medication and can closely link the safety and effectiveness of clinical medication is precisely the The Urgent Task of Modernization of Traditional Chinese Medicine

Method used

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  • A biological detection method for quality evaluation and quality control of heat-clearing and detoxifying traditional Chinese medicines
  • A biological detection method for quality evaluation and quality control of heat-clearing and detoxifying traditional Chinese medicines
  • A biological detection method for quality evaluation and quality control of heat-clearing and detoxifying traditional Chinese medicines

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Preparation of reference substance solution: Take appropriate amount of quercetin, quercetin and gallic acid reference substance, weigh them accurately, add methanol to make a mixed solution containing 15 µg of quercetin, 15 µg of quercetin and 20 µg of gallic acid per 1 ml, Instantly.

[0033] Preparation of the test sample: Take an appropriate amount of the test sample, mix it evenly, take about 0.1g, weigh it accurately, put it in a 25mL measuring bottle, add 10mL water to dissolve it (ultrasound can be used to help dissolve it if necessary), add methanol to dilute to Scale, filter (0.45μm microporous membrane), take the subsequent filtrate, that is.

[0034] 1. Chromatographic conditions

[0035] Mobile phase: A: B = acetonitrile: 0.1% phosphoric acid aqueous solution (gradient elution program as shown in Table 1), column temperature: 35 degrees, flow rate: 1.0ml / min, wavelength: 254nm.

[0036]

[0037] Accurately draw the mixed reference solution (quercetin 1...

Embodiment 2

[0040] Example 2 Characterization of anti-influenza virus activity of heat-clearing and detoxifying traditional Chinese medicines by anti-rabbit erythrocyte aggregation method

[0041] Alsever's solution: Take 0.5g of citric acid monohydrate, 8.0g of trisodium citrate dihydrate, 20.5g of glucose, and 4.2g of sodium chloride, add water to dissolve and dilute to 1L (pH about 6.2), dispense Bacteria, let cool, and store at 4°C for later use.

[0042] Preparation of 1% erythrocyte suspension: Take 8 mL of rabbit blood, mix it with an equal volume of Alfred’s solution, wash with an appropriate amount of PBS for 3 times, and wash at 2000 r min for the last time. -1 Centrifuge for 10 minutes, take 1 mL of packed red blood cells, make 100 mL of 1% red blood cell suspension with PBS, and store at 4°C for later use.

[0043] Phosphate buffered saline (PBS): Take 1.10g of anhydrous disodium hydrogen phosphate, 0.32g of anhydrous sodium dihydrogen phosphate, and 8.50g of sodium chloride...

Embodiment 3

[0057] Example 3 The method of resisting splenomegaly in mice to characterize the immune and anti-inflammatory activity of traditional Chinese medicines for clearing away heat and detoxification

[0058] Weighed 48 SPF grade C57 female mice (13-20 g), shaved their back hair with a shaver to form a skin area with a size of about 2 cm × 3 cm, and raised them in single cages. According to body weight, they were randomly divided into 6 groups, 8 in each group, respectively blank group (NG), model group (MG), high-dose group of alcohol extract of Angelica dahurica (BH, 0.112g / mL), low dose of alcohol extract of Angelica dahurica group (BL, 0.056g / mL), high-dose monkey earring water extract group (HH, 0.256g / mL) and monkey earring water extract low-dose group (HL, 0.128g / mL).

[0059] The blank group smeared 80 mg of Vaseline on the back of the mice regularly every day, and gavaged the mice with normal saline at the same time (the dosage was 0.04 mL·g -1 ), the model group applied ...

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Abstract

The invention provides a biological detection method used for quality evaluation on and quality control over heat-clearing and detoxifying traditional Chinese medicines. The biological detection method comprises the steps of firstly adopting a method of anti-rabbit erythrocytea condensation outside the body to represent anti influenza virus activity of heat-clearing and detoxifying traditional Chinese medicines, then adopting a method of anti-mouse spleen swelling method inside the body to represent immune anti-inflammatory activity of heat-clearing and detoxifying traditional Chinese medicines, comparatively completely representing activity of anti-pathogenic microorganisms and anti-inflammatory-immune regulation of heat-clearing and detoxifying medicinal materials through the combination of identification experiments inside and outside the body, and identifying the quality of the medicinal materials through a biological evaluation method. The biological detection method used for the quality evaluation on and the quality control over the heat-clearing and detoxifying traditional Chinese medicines provides a more objective quantitative evaluation method for the clinical effectiveness of the medicinal materials, and guarantees high quality and effectiveness of the medicinal materials.

Description

technical field [0001] The invention relates to the technical field of quality control of traditional Chinese medicines, in particular to a biological detection method for quality evaluation and quality control of heat-clearing and detoxifying traditional Chinese medicines. Background technique [0002] The development of identification methods of Chinese herbal medicines is synchronized with the development of traditional Chinese medicine, which is reflected in the ancient medical classics and herbal records. After the introduction of western pharmacognosy identification methods into China, four identification method systems of basic, morphology, microscopic and physical and chemical identification of traditional Chinese medicine have been formed. Then our country innovatively applied powder identification in the microscopic identification method. At present, TLC method has become the mainstream of physical and chemical identification methods of traditional Chinese medicin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02
CPCG01N33/5008G01N33/5044
Inventor 潘卫松潘超美
Owner GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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