Method for simultaneously detecting uridine and ergosterol and application thereof

A technology for ergosterol and uridine, which is applied in the field of component detection of traditional Chinese medicinal materials, can solve problems such as peak shape asymmetry, and achieve the effect of detecting a wide range of applications.

Active Publication Date: 2019-04-16
DONGGUAN HEC CORDYCEPS R&D CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a method for simultaneously detecting uridine and ergosterol, which overcomes the problems of peak collapse and peak shape

Method used

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  • Method for simultaneously detecting uridine and ergosterol and application thereof
  • Method for simultaneously detecting uridine and ergosterol and application thereof
  • Method for simultaneously detecting uridine and ergosterol and application thereof

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Experimental program
Comparison scheme
Effect test

preparation example Construction

[0044] 4. Preparation of reference solution

[0045] Weigh 20 mg of uridine and 65 mg of ergosterol respectively, weigh them accurately, put them in a 50 mL volumetric flask, add an appropriate amount of anhydrous methanol to sonicate, and after the reference substance is completely dissolved, dilute to the mark with anhydrous methanol, shake well, and A mixed stock solution of the reference substance was obtained. Before use, this reference substance stock solution was diluted 1 to 100 times with anhydrous methanol according to the needs of the experiment to make a series of mixed reference substance solutions, which were passed through a 0.45 μm organic filter membrane before use. Except for the linear investigation, the reference substance solutions used in other investigations were the dilutions of the mixed reference substance solution of uridine 0.340 mg / mL and ergosterol 0.408 mg / mL.

[0046] 5. Preparation of the test solution

[0047] Take about 0.5g of S3 sample po...

Embodiment 1

[0048] Embodiment 1: the establishment of the high-performance liquid chromatography method of the HPLC characteristic spectrum of Dictyostelium obstetricus

[0049] 1.1 Investigation of different gradient elution

[0050] Waters HSST3 chromatographic column (4.6mm×150mm, 5μm), the mobile phase is 0.1% formic acid water (A)-methanol (B), gradient elution, flow rate 1mL / min, detection wavelength 260nm, column temperature 30°C, 3 gradients For elution, inject 5 μL, 10 μL and 20 μL respectively, and the gradient elution conditions are as follows:

[0051] Gradient elution 1: 0~15min, 15%B; 15~16min, 15%→100%B; 16~30min, 100%B; 30~35min, 100%→15%B;

[0052] Gradient elution 2: 0~10min, 0%→15%B; 10~20min, 15%→30%B; 20~30min, 30%→100%B; 30~35min, 100%B; 35~40min, 100% → 0% B;

[0053] Gradient elution 3: 0~5min, 0%B; 5~15min, 0%→4%B; 15~45min, 4%→100%B; 45~60min, 100%B.

[0054] Get mixed reference substance dilution and need testing solution, analyze by above-mentioned chromato...

Embodiment 2

[0059] Embodiment 2: the extraction method investigation in the preparation of need testing solution

[0060] 2.1 Investigation of extraction solvent

[0061] Preparation of the test article:

[0062] Take about 0.5g of S3 sample powder, accurately weigh 3 parts, put it in a 10mL centrifuge tube, add 5mL ultrapure water, 50% methanol and anhydrous methanol respectively, ultrasonically extract for 30min, let cool to room temperature, take the supernatant and pass through 0.45 μm organic filter membrane, take the subsequent filtrate, that is.

[0063] Another 2 mL of anhydrous methanol extract was taken into an evaporating dish, evaporated to dryness in a water bath at 80°C, and prepared 2 parts in parallel. After evaporation, 2 mL of ultrapure water and 50% methanol were added to redissolve, and the reconstituted solution was passed through a 0.45 μm organic filter membrane. Take the continued filtrate, that is.

[0064] Chromatographic conditions:

[0065] Waters HSST3 chr...

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Abstract

The invention belongs to the field of composition detection of traditional Chinese medicinal materials, and particularly relates to a method for simultaneously detecting uridine and ergosterol and application thereof. The method comprises the following steps that a test solution is prepared; a mixed reference solution is prepared; and HPLC measurement is carried out, and specifically High Performance liquid Chromatography detection is carried out on the test solution and the mixed reference solution, wherein the sample feeding amount of the test solution is 0.5-2.5 microliters. The method overcome the defects of peak collapse, asymmetric peak shape and the like which occur when the uridine is extracted and analyzed by using an organic solvent, simultaneous detection of the uridine and theergosterol is realized, and meanwhile, the detection method is stable, reliable and wide in application range.

Description

technical field [0001] The invention belongs to the field of detection of components of traditional Chinese medicinal materials, in particular to a method for simultaneously detecting components of uridine and ergosterol. Background technique [0002] As the types of medicinal fungi continue to increase, my country has become a veritable country of edible and medicinal fungi. Edible and medicinal fungi have important medicinal value because they contain polysaccharides, polypeptide amino acids, steroids, nucleosides, alkaloids, triterpenes, minerals, vitamins and other active ingredients. So far, the research on the active ingredients of edible and medicinal fungi has mainly focused on the research on a single type of ingredients such as polysaccharides, triterpenes, nucleosides and steroids, and there are relatively few reports on the simultaneous study of multiple types of active ingredients. At present, there have been studies on the simultaneous determination of nucleos...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 钱正明黄琦李春红谢美霞朱志钢丰其娥
Owner DONGGUAN HEC CORDYCEPS R&D CO LTD
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