Method for separating and identifying bitter polypeptides in yellow rice wine

A technology for bitterness and rice wine, applied in material separation, analysis materials, measurement devices, etc., can solve the problems of result interference, high material components, complexity, etc., and achieve the effect of reducing the possibility of false positives

Pending Publication Date: 2021-01-29
ZHEJIANG SHUREN COLLEGE ZHEJIANG SHUREN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, rice wine, as a wine with Chinese characteristics, has a sugar content much higher than that of red wine and sake, and its material components are more complex. The presence of sugar will often cause great interference to the result. (Wheat) is the raw material, which is rich in protein, which makes the properties of rice wine very different from red wine and sake.

Method used

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  • Method for separating and identifying bitter polypeptides in yellow rice wine
  • Method for separating and identifying bitter polypeptides in yellow rice wine
  • Method for separating and identifying bitter polypeptides in yellow rice wine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Electronic Tongue Evaluation of Bitterness Intensity

[0040] The freeze-dried components were dissolved in 100 mL of deionized water, centrifuged at 3500 rpm for 10 min, and 80 mL of the supernatant was taken for electronic tongue detection. First, wash in the cleaning solution for 90s, then wash with the reference solution for 120s, then wash with another reference solution for 120s, the sensor returns to zero at the equilibrium position for 30s, after reaching equilibrium, soak in the separated component for 30s for measurement. Do 4 cycles for each sample, remove the first cycle, and take the average value of the last 3 cycles. And complete data processing in TS-5000Z in the database of its instrument terminal system server to obtain relative bitterness intensity.

Embodiment 2

[0041] Example 2: Preliminary Separation of Yellow Rice Wine Samples

[0042] (1) Separation of rice wine samples by distillation under reduced pressure

[0043] Take 300mL of rice wine and carry out vacuum distillation (conditions of vacuum distillation: -0.1MPa, 45°C, 90rpm), when the volume of rice wine is less than 30mL, stop the rotary evaporation, and freeze-dry the remaining low-volatile components to obtain difficult Volatile component F was stored at -20°C for later use, and its bitterness intensity was evaluated by electronic tongue.

[0044] (2) ethanol alcohol precipitation to separate difficult volatile components F

[0045] Take the less volatile component F, dissolve it with 20mL of water, slowly add ethanol so that the final concentration of ethanol is 80%, and keep shaking and mixing, and let it stand at 4°C for 12h to obtain the ethanol phase, alcohol precipitation phase, and ethanol phase Distilled under reduced pressure (-0.1Mpa, 45°C, 90rpm), freeze-drie...

Embodiment 3

[0054] Example 3: Separation of F-1-1-Ⅲ by semi-preparative HPLC

[0055] F-1-1-III (200 mg) was dissolved in 20 mL of water, filtered through a membrane (0.45 μm), and separated by semi-preparative HPLC.

[0056] Column: Xbridge Prep C 18 (5μm, 10mm×250mm); mobile phase: A ultrapure water, B methanol; elution conditions: 0min A / B (80:20), 5min A / B (70:30), 15min A / B (70: 30), 16min A / B(50:50), 25min A / B(50:50), 26min, A / B(30:70), 31min A / B(30:70), 32min A / B(0 :100), 40min, A / B(0:100), 41min A / B(80:20), 45min A / B(80:20); flow rate: 4mL / min; detection wavelength: 272nm; injection volume: 600 μL. According to the chromatographic peaks, fractions F-1-1-Ⅲ were divided into 16 fractions, respectively denoted as III-1~Ⅲ-16, and the same fractions were combined, lyophilized after rotary evaporation, and stored at -20°C for later use .

[0057] Chromatographic results such as figure 2 As shown, by semi-preparing C 18 The chromatographic column divides F-1-1-III into 16 compone...

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Abstract

The invention discloses a method for separating and identifying bitter polypeptide in yellow rice wine, and belongs to the technical field of food. After electronic tongue determination is combined with ethanol precipitation, n-butyl alcohol extraction, C18SPE solid-phase extraction and semi-preparative HPLC separation, the bitter polypeptide in the yellow rice wine is analyzed, identified and separated by combining amino acid component analysis, UPLC-ESI-Q-TOF-MS / MS, pyroglutamate peptidase digested polypeptide and mass spectrum software, and finally, the polypeptide Pyr-LFNPSTNPWHSP with bitterness is identified from the yellow rice wine. According to the method, multiple methods are combined to separate and identify the bitter polypeptide in the yellow rice wine, pyroglutamic acid peptidase digestion verifies that the polypeptide is the bitter polypeptide containing pyroglutamic acid, and a theoretical basis is provided for research of a mechanical yellow rice wine bitter taste reducing technology; and through pyroglutamate peptidase digestion verification and other identification methods, the false positive possibility of determining the polypeptide sequence only by a mass spectrometry MS / MS method can be effectively reduced.

Description

technical field [0001] The invention relates to a method for separating and identifying bitter polypeptides in yellow rice wine, belonging to the technical field of food. Background technique [0002] Mechanized rice wine has been developed for more than 30 years, and its technology is becoming more and more perfect. Mechanized large-tank fermented rice wine has the advantages of small footprint, relatively stable product quality, no seasonal restrictions on production, greatly reduced labor intensity, and more hygienic and safer products. Especially in recent years, due to the widespread problem of difficult employment in rice wine enterprises, the development of mechanized production has become an inevitable choice for rice wine enterprises. [0003] Bitterness is one of the basic tastes of rice wine. Appropriate bitterness gives rice wine a strong and refreshing feeling, but too much bitterness will destroy the coordination of the wine and make the wine body impure. How...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/12
CPCG01N30/02G01N30/06G01N30/12G01N2030/062G01N2030/126
Inventor 谢广发陆健吴殿辉鲁振东胡志明许锡飚陈伟峰
Owner ZHEJIANG SHUREN COLLEGE ZHEJIANG SHUREN UNIV
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