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Method for quickly determining six soy isoflavones in bean product by utilizing UPLC (Ultra Performance Liquid Chromatography)

A technology for rapid determination of soybean isoflavones, applied in the field of analytical chemistry, can solve the problems of fewer reports of qualitative and quantitative soybean isoflavones, and achieve the effects of low cost, simple pretreatment, accurate identification and evaluation

Inactive Publication Date: 2015-01-07
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the detection of soybean isoflavones is mainly concentrated in soybeans, and there are few reports on the qualitative and quantitative analysis of soybean isoflavones in soybean products

Method used

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  • Method for quickly determining six soy isoflavones in bean product by utilizing UPLC (Ultra Performance Liquid Chromatography)
  • Method for quickly determining six soy isoflavones in bean product by utilizing UPLC (Ultra Performance Liquid Chromatography)
  • Method for quickly determining six soy isoflavones in bean product by utilizing UPLC (Ultra Performance Liquid Chromatography)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Determination of soybean isoflavones in embodiment 1 soybean milk

[0019] 1. Standard samples: daidzin, daidzein, daidzein, glycitin, genistin, and genistein were purchased from Aladdin Company.

[0020] 2. Sample pretreatment: Take 0.1 mL of soybean milk and add 0.9 mL of methanol, ultrasonically extract at 53 kHz for 5 min, vortex and mix well, centrifuge at room temperature at 4000 r / min for 3 min, take the supernatant, and repeat extraction of the residue twice with methanol , combined supernatants, blown dry with nitrogen, passed through a 0.22 μm filter membrane, and tested.

[0021] 3. Chromatographic conditions: Chromatographic column: Kinetex-C18 (2.1 mm×100 mm, 1.7 μm), mobile phase is acetonitrile phase A and 0.1 % formic acid phase B, flow gradient elution program: 0-0.5 min: 10 % A+ 90% B; 0.5-10 min: 10% A+90% B - 45% A+55% B; 10-13 min: 45% A+55% B - 100% A;

[0022] The flow rate was 0.3 mL / min, the column temperature was 45 °C, and the UV detection w...

Embodiment 2

[0025] Example 2 Determination of soybean isoflavones in tofu

[0026] 1. Standard samples: daidzin, daidzein, daidzein, glycitin, genistin, and genistein were purchased from Aladdin Company.

[0027] 2. Sample pretreatment: Take 0.1g of tofu and add 1 mL of methanol, ultrasonically extract at 53kHz for 5 min, vortex and mix well, centrifuge at room temperature at 4000 r / min for 3 min, take the supernatant, and repeat the extraction of the residue twice with methanol , combined supernatants, blown dry with nitrogen, passed through a 0.22 μm filter membrane, and tested.

[0028] 3. Chromatographic conditions: Chromatographic column: Kinetex-C18 (2.1 mm×100 mm, 1.7 μm), mobile phase is acetonitrile (phase A) and 0.1% formic acid (phase B), flow gradient elution program: 0-0.5 min: 10 % A+90 % B; 0.5-10 min: 10 % A+90% B - 45% A+55% B; 10-13 min: 45% A+55% B - 100 % A; flow rate 0.3 mL / min, column temperature 45 ℃, UV detection wavelength 260 nm; injection volume 1 μL.

[002...

Embodiment 3

[0031] Determination of Soybean Isoflavones in Example 3 Vegetarian Chicken

[0032] 1. Standard samples: daidzin, daidzein, daidzein, glycitin, genistin, and genistein were purchased from Aladdin Company.

[0033] 2. Sample pretreatment: take 0.1g of vegetarian chicken and add 1 mL of methanol, ultrasonically extract at 53kHz for 5 min, vortex and mix well, centrifuge at room temperature at 4000 r / min for 3 min, take the supernatant, and repeat the extraction of the residue with methanol 2 For the second time, the supernatants were combined, blown dry with nitrogen, and passed through a 0.22 μm filter membrane for testing.

[0034] 3. Chromatographic conditions: Chromatographic column: Kinetex-C18 (2.1 mm×100 mm, 1.7 μm), mobile phase is acetonitrile phase A and 0.1 % formic acid phase B, flow gradient elution program: 0-0.5 min: 10 % A+ 90 %B; 0.5-10 min: 10 % A+90% B - 45% A+55% B; 10-13 min: 45% A+55% B - 100 % A; flow rate 0.3 mL / min, column temperature 45 ℃, UV detecti...

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Abstract

The invention discloses a method for quickly determining six soy isoflavones in a bean product by utilizing UPLC (Ultra Performance Liquid Chromatography), and belongs to the technical field of analytical chemistry. The types of the tested bean product comprise soybean milk, bean curd, soy chicken, dried tofu, bean curds, dried beancurd sticks and the like, and the six soy isoflavones such as daidzin, isoflavoues Aglycone, Glycitin, glycitein, genistin and genistein are determined. The method comprises the following steps: firstly extracting the soy isoflavones in the bean product by using a methanol solution, after collecting a supernate fluid, repeatedly extracting residual twice by using methanol, merging the supernate fluids, and carrying out UPLC detection. Quick determination for the composition and content of the six soy isoflavones mainly in the bean product are achieved by adopting UPLC. The method is simple in sample pretreatment, high in analysis speed, high in repeatability and accuracy, and provides an efficient and accurate analytical method means for evaluating and controlling the quality of the bean product.

Description

technical field [0001] The invention discloses a method for rapidly determining six kinds of soybean isoflavones in soybean products by using UPLC, belonging to the technical field of analytical chemistry. Specifically design a method of using UPLC (Ultra High Performance Liquid Chromatography) to determine daidzin, daidzein, daidzein, daidzein, genistin, Genistein 6 Methods of Soy Isoflavones. Background technique [0002] Soybean isoflavones refer to a series of compounds with 3-benzopyrone as the core, including three free aglycones and bound glycosides. Glycosidic soybean isoflavones include four types: glucoside, acetylated glycoside and malonylated glycoside. Compared with isoflavone glucoside, isoflavone aglycone has more extensive and obvious biological activity and higher absorption and utilization rate. Soy isoflavones are mainly derived from the pods of legumes, among which soybeans have the highest content. They are a kind of natural estrogen, which ha...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 张爽陈洁陶冠军秦昉曾茂茂郑宗平何志勇
Owner JIANGNAN UNIV
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