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HPLC-FLD detection method of radix pseudostellariae homogeneous polysaccharide

A technology of HPLC-FLD and detection method, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of unclear absorption, transport, delivery and other mechanisms of polysaccharide molecular forms, and detection difficulties, etc., to achieve accurate methods, The method is simple and has broad application prospects

Pending Publication Date: 2022-04-05
福建省中医药科学院
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Problems solved by technology

[0003] Due to the detection difficulties caused by the lack of chromophores of polysaccharides (very weak absorption under UV conditions), the research on carbohydrate molecules has been unable to break through in the field of biochemistry. The molecular form of polysaccharides in the body is absorbed, transported, and delivered. very clear

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  • HPLC-FLD detection method of radix pseudostellariae homogeneous polysaccharide
  • HPLC-FLD detection method of radix pseudostellariae homogeneous polysaccharide
  • HPLC-FLD detection method of radix pseudostellariae homogeneous polysaccharide

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Embodiment Construction

[0014] In order to make the content of the present invention easier to understand, the technical solutions of the present invention will be further described below in conjunction with specific embodiments, but the present invention is not limited thereto.

[0015] 1. Pseudostellaria heterogenea polysaccharide (PHP) 2-AB fluorescent labeling

[0016] 1. Preparation of 2-AB-PHPs

[0017] Take 350 μL of DMSO to dissolve 350 mg of TBAF, and add 120 μL of Ac. After mixing, add 5mg of 0.5M polysaccharide, vortex for 30min, dissolve at 65°C for 1h, until clear; add 20mg of 2-AB, mix; add 10mg of sodium cyanoborohydride, mix well, and incubate at 65°C for 4h. TLC see figure 1 , see the chemical structure figure 2 a.

[0018] 2. 2-AB-dextran preparation

[0019] Take 350 μL of DMSO and add 150 μL of Ac to mix. Dissolve 25mg of 2-AB with the mixed solution, add 30mg of sodium cyanoborohydride, mix well, add 5mg of dextran, and incubate at 65°C for 4h; treatment of labeled polysac...

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Abstract

The invention discloses an HPLC-FLD (High Performance Liquid Chromatography-FLD) detection method of radix pseudostellariae homogeneous polysaccharide, which comprises the following steps: carrying out fluorescence labeling on the radix pseudostellariae homogeneous polysaccharide by using 2-aminobenzidine, quantitatively analyzing the radix pseudostellariae homogeneous polysaccharide by using glucan as a standard substance and HPLC-FLD, and determining the radix pseudostellariae homogeneous polysaccharide in a biological sample cell culture solution. The method is simple, sensitive, accurate and wide in application prospect.

Description

technical field [0001] The invention belongs to the technical field of fluorescence detection of polysaccharides, and in particular relates to an HPLC-FLD detection method of heterogeneous polysaccharides from Radix Pseudostellariae. Background technique [0002] Polysaccharides are one of the three major substances that constitute life sciences and are widely found in traditional Chinese medicine. Polysaccharides have various biological activities such as immune regulation, anti-tumor, hypoglycemic, hypolipidemic, anti-oxidation, anti-aging, anti-virus, etc., and pharmacological studies have been widely confirmed. [0003] Due to the detection difficulties caused by the lack of chromophores of polysaccharides (very weak absorption under UV conditions), the research on carbohydrate molecules has been unable to break through in the field of biochemistry. The molecular form of polysaccharides in the body is absorbed, transported, and delivered. very clear. [0004] The detec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/74
Inventor 胡娟庞文生杨斌栗园陈锦龙阚永军
Owner 福建省中医药科学院