Method of quickly screening standard substance used in quantitative detection of trifolium pratense isoflavone

A technology for the quantitative detection of red clover isoflavones, which is applied in the field of rapid reduction and screening of standard products used in the quantitative detection of red clover isoflavones, can solve problems affecting the development of the red clover industry and achieve low detection costs and fast screening , The effect of simple process

Inactive Publication Date: 2018-06-22
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no report on how to select standard products and the related errors caused by them, which will affect the further development of the red clover industry

Method used

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  • Method of quickly screening standard substance used in quantitative detection of trifolium pratense isoflavone
  • Method of quickly screening standard substance used in quantitative detection of trifolium pratense isoflavone
  • Method of quickly screening standard substance used in quantitative detection of trifolium pratense isoflavone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Effect of Different Standard Items on Spectrophotometric Determination of Red Clover Isoflavones Results

[0016] 1. Preparation of red clover samples: Three red clover samples were randomly collected, air-dried and pulverized, and named samples 1, 2 and 4. Take 0.2 g of naturally air-dried red clover leaves, soak them in 8 mL of methanol, and extract them by ultrasonic wave for 30 minutes. After centrifugation for 10 minutes, the supernatant was taken and filtered through a 0.45 micron organic membrane to prepare the test solution.

[0017] 2. Preparation of standard products:

[0018] Weigh 5mg of formononetin reference substance and place it in a 100mL volumetric flask, add methanol to dilute to the mark to obtain a formononetin reference substance solution with a concentration of 0.05mg / mL.

[0019] Weigh 10mg of genistein standard substance into a 100mL volumetric flask, and then add methanol to calibrate to the scale to obtain a genistein standard solu...

Embodiment 2

[0030] Embodiment 2 Thin-layer chromatography screening standard method

[0031] Using a capillary tube, spot the mixed samples of the four standard products and the samples of red clover 1, 2 and 4 to be tested on the prepared silica gel GF254 thin-layer chromatography plate, put them into a developing bottle, and use chloroform:methanol= 15:1 is the developing agent for developing. When the developing agent reaches 1-2 mm from the edge of the TLC plate, take out the chromatographic plate, dry it, and observe it under a 254nm ultraviolet lamp.

[0032] Such as figure 2 As shown, in the figure B represents garbanzoin A; F represents formononetin; G represents genistein; D represents daidzein. The spots of samples 1 and 2 formononetin (standard product F) are more obvious, while the spots of sample 4 chickpein A (standard product B) are more obvious.

[0033] Based on the results, it is suggested that samples 1 and 2 use formononetin or daidzein as the standard, and sample ...

Embodiment 3

[0034] Embodiment 3 Thin-layer chromatography screens standard substance method

[0035] Because the standard substances chickpein A and formononetin have strong ultraviolet absorption, only these two standard substances can be used to complete the screening.

[0036] Using a capillary tube, the mixed sample mixed with the standard chickpein A and formononetin and the red clover 1, 2 and 4 samples to be tested were spotted on the prepared silica gel GF254 thin-layer chromatography plate to Chloroform:methanol=20:1 for development. When the developing agent reaches 1-2 mm from the edge of the TLC plate, take out the chromatographic plate, dry it, and observe it under a 254nm ultraviolet lamp.

[0037] Such as figure 2 As shown, the spots of formononetin (standard substance F) in samples 1 and 2 are more obvious, while the spots of chickpein A (standard substance B) in sample 4 are more obvious.

[0038] Based on the results, it is suggested that samples 1 and 2 use formonon...

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Abstract

Trifolium pratense isoflavone is an important phytoestrogen, has an important clinical medicinal value, is an important biochemical ingredient of a plant to cope with adversity stress, and is closelyrelated to various factors such as an environment, a growth cycle and plant after-treatment. Accurate quantitation of isoflavone is a critical foundation of research and applications; however, standard substances used for a quantitation process of trifolium pratense isoflavone are inconsistent at present; relevant studies on errors arising from the inconsistent standard substances are absent. Theinvention discloses a method which adopts 2-4 compounds for thin-layer chromatography calibration of a plant sample on a basis of summarized ultraviolet absorption rules of major compounds of trifolium pratense isoflavone so as to quickly screen the standard substances used in the plant sample to be determined. Via verification by high performance liquid chromatography, the method is scientific and reliable, simple, mature and reliable in technology, low in detection cost, and easy to realize outside a laboratory, saves time and labor, and has low requirements on a detection environment, goodextensibility and huge application and development prospects.

Description

technical field [0001] The invention belongs to the field of medicine and food quality inspection, in particular to a method for rapidly reducing and screening standard substances used in the quantitative detection of red clover isoflavones. Background technique [0002] Red clover (Trifolium pratense L.), also known as red clover, red clover, etc., is an excellent legume forage with a wide distribution. Because red clover contains high content of isoflavones, it is internationally recognized as a kind of herbal medicine with relatively accurate curative effect on women's climacteric syndrome, and its sales volume has been ranked among the top 10 herbal medicines in the United States for many years. In addition, studies have shown that excessive red clover isoflavones can also lead to hormonal disorders in eating livestock. [0003] The total isoflavone content of red clover is mainly carried out by ultraviolet spectrophotometry. Generally, 1-2 standard products are used t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/90
CPCG01N30/90
Inventor 刘权蔺永和李晓蓓
Owner LANZHOU UNIVERSITY
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