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Method used for resolution of equol enantiomers, and measuring of content of equol enantiomers in bean products

A technology for enantiomers and soybean products, applied in the field of biochemical analysis and detection, can solve the problems of high pretreatment requirements, inability to detect a single enantiomer, complicated steps, etc., and achieve high sensitivity and enhanced ionization efficiency. , the effect of simplifying the extraction process

Active Publication Date: 2016-12-21
SHANDONG ANALYSIS & TEST CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, enzyme-labeled immunosorbent assay and gas chromatography cannot detect a single enantiomer, while high-performance liquid chromatography has high requirements for sample pretreatment, cumbersome steps, and there may be interference of false positives

Method used

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  • Method used for resolution of equol enantiomers, and measuring of content of equol enantiomers in bean products
  • Method used for resolution of equol enantiomers, and measuring of content of equol enantiomers in bean products
  • Method used for resolution of equol enantiomers, and measuring of content of equol enantiomers in bean products

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1 Experimental part

[0039] 1.1 Instruments and reagents

[0040]The main instruments used in the experiment are: Agilent-1200 type rapid separation high performance liquid chromatography (Agilent Company), 6410 type triple quadrupole mass spectrometer (Agilent Company). Methanol (chromatographically pure, Tedia Company), ammonium acetate (analytical pure, Sinopharm Group) were both chromatographically pure. The racemic compound of equol and S-equol were purchased from Sigma Company (purity greater than 98%). Chloramphenicol-D5 (internal standard IS) was purchased from AccuStandard Company (purity 99%, USA); CHIRALCEL OJ-RH chiral column (Daicel Company, Japan).

[0041] 1.2 HPLC-MS / MS conditions

[0042] 1.2.1 HPLC conditions

[0043] Chromatographic column: CHIRALCEL OJ-RH chiral column (150mm×4.6mm, 5μm, Japan Daicel Company). Mobile phase: Phase A is an aqueous solution containing 10 mmol / L ammonium acetate, and phase B is methanol. Isocratic elution: A:B=80:...

Embodiment 2

[0059] Embodiment 2 linear relationship and detection limit

[0060] Take an appropriate amount of equol racemic compound standard solution and chloramphenicol-D5 standard solution and add it to the blank fermented bean curd sample, and accurately prepare a series of samples with different mass concentrations of 5.0, 20, 100, 200, 500, and 1000 μg / kg. The method specified under "item 1.3 in embodiment 1" is processed, and under optimized conditions, the sample injection is measured respectively, and the quantitative ion peak area average value of the equol enantiomer is used for the internal standard quantitative ion peak area respectively. The ratio is the ordinate (Y), and the linear regression is carried out on the abscissa (X) with the equol enantiomer concentration and the internal standard concentration ratio respectively (the linear graph is shown in Figure 3-Figure 4 ), get the regression equation, correlation coefficient and linear range; take the concentration corre...

Embodiment 3

[0063] Embodiment 3 standard addition recovery rate

[0064] Add equol racemic standard solution to the blank fermented bean curd sample, so that the concentration of the blank spiked sample is 200 μg / kg, process according to the method specified under "1.3", repeat the measurement 5 times, calculate the recovery rate and precision, and the result It shows that the recovery rate and precision of equol enantiomers all meet the requirements of "Laboratory Quality Control Standards for Physical and Chemical Testing of Food" (GB / T 27404-2008), and the results are shown in Table 3.

[0065] Table 3. The recovery and precision test results of equol enantiomers added to blank sufu samples (n=5)

[0066]

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Abstract

The invention discloses a method used for resolution of equol enantiomers, and measuring of the content of equol enantiomers in bean products. The method comprises following steps: 1, bean products to be measured are subjected to pretreatment, wherein methanol extraction and adding of internal standards are adopted; 2, after pretreatment, high performance liquid chromatography-tandem mass spectrometry is adopted to separate and measure equol enantiomers in the bean products to be measured, wherein high performance liquid chromatography conditions are determined as following, a polysaccharide derivative reversely-coated chiral chromatographic column is taken as a chromatographic column, a mobile phase is composed of an A phase and a B phase, the A phase is an ammonium acetate aqueous solution, and the B phase is methyl alcohol. According to the method, high performance liquid chromatography-tandem mass spectrometry is adopted to separate equol enantiomers, optimal analysis conditions are determined via optimization of liquid chromatography parameters, extraction process is simplified, influences of matrix effects are avoided via adding of the internal standards, and the method is capable of avoiding false positive results effectively, and realizing rapid measuring of S-equol in bean products.

Description

technical field [0001] The invention belongs to the field of biochemical analysis and detection, in particular to a method for splitting equol enantiomers and measuring their content in soybean products. Background technique [0002] Equol, the chemical name is 7-hydroxy-3-(4-hydroxyphenyl)-chroman, the chemical structure is as follows, it is produced by the metabolism of soybean isoflavones in the human intestinal tract by certain bacteria One of the final metabolites. In recent years, studies have found that equol is the main embodiment of the physiological functions of soybean isoflavones, and has a role in diseases related to estrogen regulation, such as breast cancer, prostate cancer, menopausal syndrome, cardiovascular diseases, and osteoporosis. Preventive and therapeutic effects. However, it has been found that only 30%-50% of human individuals can metabolize soybean isoflavones into equol under the action of their intestinal microbial flora. Therefore, exogenous s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/062
Inventor 苑金鹏王珊珊李福伟王晓利赵汝松陈跃
Owner SHANDONG ANALYSIS & TEST CENT
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