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A rapid and efficient method for determining the content of cysteine ​​and glutathione in horsetail

A technology for glutathione content and cysteine, which is applied in the field of analysis and determination, can solve the problems of high cost and long detection time, and achieve the effect of reducing consumption cost and improving detection efficiency

Inactive Publication Date: 2017-11-28
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to solve the problems of high cost and long detection time in the existing Cys and GSH determination methods, the present invention provides a method for fast and efficient qualitative and quantitative analysis of Cys and GSH contents in S.

Method used

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  • A rapid and efficient method for determining the content of cysteine ​​and glutathione in horsetail
  • A rapid and efficient method for determining the content of cysteine ​​and glutathione in horsetail
  • A rapid and efficient method for determining the content of cysteine ​​and glutathione in horsetail

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Experimental program
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Effect test

Embodiment 1

[0031] (1) Sample processing and extraction: Iris seedlings with a plant height of 15-20 cm were subjected to 0, 100, 500 mg·L -1 Pb(NO 3 ) 2 After 7 days of treatment, Cys and GSH were extracted from the leaves and roots of Iris and derivatized. The details are as follows: Weigh 0.3 g of fresh Iris sativa tissue, quickly freeze it in liquid nitrogen, place it in a mortar, add 1.75 ml of 0.1% TFA (containing 6.3 mmol·L -1 DTPA, pH <1) and a small amount of quartz sand, fully ground and centrifuged on an ice bath (4 ℃, 12000g, 20min), and the supernatant was refrigerated for analysis and determination of Cys and GSH.

[0032] (2) Standard solution configuration: Accurately weigh 0.1g of each of Cys and GSH, dissolve them in 1ml of 0.1% TFA, use them as standard stock solutions, and store them at -20°C for later use; before use, dilute them with 0.1% TFA to 1, 5, 10, 20, 100ng·µl -1 Standard series, store at 4°C.

[0033] (3) Derivatization: Take 25µl standard solution and ...

Embodiment 2

[0041] (1) Sample processing and extraction: Iris seedlings with a plant height of 15-20 cm were treated with 0, 10, 80 mg·L -1 CdCl 2After 7 days of treatment, Cys and GSH were extracted from leaves and roots and derivatized. The details are as follows: Weigh 0.3 g of fresh Iris sativa tissue, quickly freeze it in liquid nitrogen, place it in a mortar, add 1.75 ml of 0.1% TFA (containing 6.3 mmol·L -1 DTPA, pH <1) and a small amount of quartz sand, fully ground and centrifuged on an ice bath (4 ℃, 12000g, 20min), and the supernatant was refrigerated for the analysis and determination of Cys and GSH.

[0042] (2) Standard solution configuration: Accurately weigh 0.1g of each of Cys and GSH, dissolve them in 1ml of 0.1% TFA, use them as standard stock solutions, and store them at -20°C for later use; before use, dilute them with 0.1% TFA to 1, 5, 10, 20, 100ng·µl -1 Standard series, store at 4°C.

[0043] (3) Derivatization: Take 25µl standard solution and plant tissue supe...

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Abstract

The invention discloses a method for quickly and efficiently measuring the content of cysteine ​​and glutathione in horsetail, which belongs to the technical field of analysis and determination. In this study, ultra-high performance liquid chromatography (UPLC) was used to qualitatively and quantitatively analyze cysteine ​​(Cys) and glutathione (GSH) in the extract of Iris japonica: Vydac C18 column was selected, and the mobile phase was 0.1% TFA (A) and 100% ACN (B); column temperature 30°C, flow rate 0.5ml min-1, injection volume 1μl; gradient elution program: 12%~25% ACN (3min), 25% ~100% ACN (0.5min); column wash: 100% ACN (1.5min); column equilibration: 100%~0% ACN (0.5min) and post-column flow time 1.5 min. The conditions of the fluorescence detector are: λex (excitation wavelength) 380 nm, λem (emission wavelength) 470 nm. The external standard method is used to quantify the peak area, and Cys and GSH can be detected and analyzed simultaneously within 7 minutes. Compared with the traditional high-performance liquid chromatography (HPLC), the method greatly shortens the analysis process, has good separation effect, greatly improves the detection efficiency and saves the mobile phase cost.

Description

technical field [0001] The invention discloses a method for quickly and efficiently measuring the content of cysteine ​​and glutathione in horsetail, which belongs to the technical field of analysis and determination. Background technique [0002] Cysteine ​​(Cys) and glutathione (GSH) are an important class of non-protein sulfhydryl compounds. Due to the sulfhydryl (-SH) contained in the compound structure, Cys and GSH play an important role in the detoxification of heavy metals. Ma Lin ( Iris lactea var chinensis ) is a perennial herb with wide adaptability, strong resistance, large biomass, and extensive cultivation and management. It has been widely used in landscaping, saline-alkali land improvement, water and soil conservation, and remediation of heavy metal-contaminated soil. Iris has a strong ability to accumulate and tolerate heavy metals, and the synthesis of non-protein sulfhydryl Cys and GSH under heavy metal stress is an important mechanism for its detoxifica...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/88
Inventor 原海燕黄苏珍杨永恒
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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