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Extracting and detecting method for flavonoid in radix sophorae subprostratae

A technology of flavonoids and detection methods, which is applied in the field of analysis and detection methods of flavonoids in root medicinal materials, can solve the problems of environmental pollution, large amount of eluent, etc., and achieve high extraction efficiency, low cost, and high efficiency.

Active Publication Date: 2017-05-31
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the matrix solid phase dispersion extraction techniques reported in the literature use florisil, diatomaceous earth, C 18 Reverse silica gel, alumina, etc. are used as adsorbents, and the amount of medicinal materials, adsorbents, and eluents is relatively large, especially if the amount of eluent is more than 10mL, it will cause pollution to the environment

Method used

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  • Extracting and detecting method for flavonoid in radix sophorae subprostratae
  • Extracting and detecting method for flavonoid in radix sophorae subprostratae
  • Extracting and detecting method for flavonoid in radix sophorae subprostratae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1: Sample pretreatment

[0068] 1. High performance liquid chromatography conditions: Agilent Technologies 1290 Infinity. Column: EclipsePlus C 18 RRHD, 1.8μm, 2.1×50mm, detection wavelength: 280nm, column temperature: 40℃, injection volume: 0.8uL, flow rate: 0.4mL min -1 , mobile phase: A: acetonitrile, B: 0.1% formic acid in water. Gradient elution: 0~30min, 10%~40%A; 30~40min, 40%~50%A; 40~50min, 50%~56%A; 50~60min, 56%~76%A; 60~ 70min, 76%~95%A; 70~75min, 95%~95%.

[0069] 2. Optimization of extraction conditions: investigate different adsorbents C 18 、Al 2 o 3 , silica gel, diatomaceous earth, florisil, ZSM-5, TS-1, SAPO-11, MCM-48, SBA-15, the ratio of different mountain bean root medicinal powder and molecular sieve is 2:1, 3:2, 1:1, 1:2, 1:3, different grinding time 90s, 120s, 150s, 180s, 210s, different eluents methanol, ethanol, acetonitrile, acetone, 50% methanol-50% acetonitrile, 50% methanol-50 Effects of % ethanol and different volumes o...

Embodiment 2

[0077] Embodiment 2: Separation condition optimization

[0078]1. Capillary Electrokinetic Chromatography Conditions: Agilent Technologies 7100Capillary Electrophoresis, with UV detector; Column: Uncoated Quartz Capillary (56cm×50μm, i.d., Hebei Yongnian Ruifeng Chromatographic Device Co., Ltd.). The operating voltage of reverse microemulsion capillary electrokinetic chromatography is -15~-30kv (preferably -20kv), the detection temperature is 20~35°C (preferably 25°C), the conventional voltage sampling conditions: -20kv, 5s; the detection wavelength is 280nm. Microemulsion consists of: SDS 0.8% ~ 3.0% (w / v), co-surfactant 6.6% (w / v), oil phase 0.5% (w / v), organic additives 15% (w / v), hand Resolving agent 0~20mM, buffer 20mM pH 2NaH 2 PO 4 aqueous solution. More preferably: 1% (w / v) SDS, 6.6% (w / v) n-butanol, 0.5% (w / v) ethyl acetate, 15% (w / v) acetonitrile, 10 mM sulfobutyl-β - Sodium cyclodextrin, 20mM NaH 2 PO 4 (pH 2) buffer, form a microemulsion after ultrasonication...

Embodiment 3

[0092] Example 3: Optimization of FASI-MEEKC online enrichment conditions

[0093] 1. The influence of the pH value and concentration of the sample matrix

[0094] The degree of dissociation of the analytes will significantly affect the enrichment effect of FASI when electrodynamic sampling is used. Flavonoids are difficult to dissociate under neutral or acidic conditions, and the enrichment effect is poor. Therefore, an alkaline sample matrix is ​​selected. The present invention investigates the borax buffer solution with pH 7.5-9.5 as the sample matrix, and finds that the peak shape is good and the enrichment effect is the best at pH 8.5.

[0095] The concentration of borax buffer affects the ionic strength and conductivity of the sample matrix. The influence of the sample matrix concentration from 0.1 to 10mol / L pH 8.5 on the enrichment effect was investigated. The results show that the detection sensitivity of the sample is obviously improved in the lower concentration so...

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Abstract

The invention provides an extracting and detecting method for flavonoid in radix sophorae subprostratae. The flavonoid to be detected is lupinifolin and radix sophorae subprostratae chroman flavanone C which are respectively called flavone 1 and flavone 4 for short. The extracting and detecting method disclosed by the invention creatively combines a matrix solid-phase dispersion extraction technology with a reverse micro-emulsion electrokinetic chromatography technology, utilizes combination of an offline enrichment technology and an online enrichment technology to achieve secondary enrichment of the flavonoid in the radix sophorae subprostratae and is beneficial to quantitative detection of the flavonoid in the radix sophorae subprostratae. Furthermore, the extracting and detecting method disclosed by the invention is implemented in a microscale system, the used medical materials and experiment drug reagents are all in a microgram level, the use amount of harmful chemical reagents is small, and safety, economical performance and environmental friendliness are achieved.

Description

[0001] (1) Technical field [0002] The invention discloses an analysis and detection method for extracting and separating flavonoids in the medicinal material of mountain bean root by combining molecular sieve matrix solid-phase extraction technology (MSPD) and reversed microemulsion capillary electrokinetic chromatography (MEEKC) online enrichment technology, specifically In other words, it involves the use of molecular sieve SBA-15 as an adsorbent, combined with the off-line enrichment of matrix solid-phase extraction technology and the on-line enrichment of reverse microemulsion field scale-up sampling (FASI-MEEKC), aiming to establish a method for Sophora japonica A method for the separation and analysis of small polar free flavonoids with prenyl side chains that are abundant in plants of the genus. [0003] (2) Background technology [0004] Sophora tonkinensis is the dry root and rhizome of Sophora tonkinensis Gagnep., a vine-like dwarf shrub of the leguminous family, ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/08
CPCG01N30/06G01N30/08G01N2030/062
Inventor 李行诺郑丽琼颜继忠
Owner ZHEJIANG UNIV OF TECH
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