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Derivatization combined LC-MS method for determining concentration of allantoin in cell matrix

A LC-MS, cell matrix technology, applied in the field of analytical chemistry, can solve the problems of inability to detect endogenous substances and low sensitivity, and achieve the effect of avoiding matrix effects, high sensitivity, and making up for poor sensitivity.

Active Publication Date: 2019-12-17
WUXI JIMIN KEXIN SHANHE PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Another method is derivatization combined with HPLC method, which can perform quantitative detection at 360nm, but this method has low sensitivity and cannot detect trace amounts of endogenous substances

Method used

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  • Derivatization combined LC-MS method for determining concentration of allantoin in cell matrix
  • Derivatization combined LC-MS method for determining concentration of allantoin in cell matrix
  • Derivatization combined LC-MS method for determining concentration of allantoin in cell matrix

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Experimental program
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Effect test

Embodiment 1

[0059] In order to confirm the accuracy of determining the concentration of allantoin in cells by derivatization combined with LC-MS, the present invention firstly prepared a series of calibration samples and quality control samples for the standard curve, and investigated whether the inventive method meets the requirements of analytical methodology. requirements, and to determine the content of endogenous glyoxylate in the cell matrix.

[0060] Reagents used: standard allantoin (China Institute for Food and Drug Control, purity>99%); 2,4-dinitrophenylhydrazine (purity≥99%, Adamas-beta); 5-nitroindole-2 - formic acid (purity ≥ 98%, source leaf organisms); acetonitrile, methanol (chromatographically pure, Tedia Corporation, USA).

[0061] Prepare allantoin, internal standard, and derivatization reagent solutions respectively:

[0062] Allantoin standard curve working solution and quality control sample working solution: Accurately weigh 15.81mg of allantoin into a 100ml measur...

Embodiment 2

[0070] The apparent permeability coefficient (P app ).

[0071] Allantoin aluminum is dissolved in water and decomposed into allantoin and aluminum hydroxide, so allantoin is used as the measurement object. It is used to treat gastric and duodenal ulcers, chronic gastritis, antral gastritis, and drug-induced gastritis; it can also be made into external preparations for treating ulcers and wounds, which can promote their healing. The structural formula of aluminum allantoin is as follows:

[0072]

[0073] Reagents used: aluminum allantoin raw material medicine (provided by Wuxi Jimin Kexin Shanhe Pharmaceutical Co., Ltd., purity > 99%); allantoin raw material medicine (China Institute for Food and Drug Control, purity > 99%);

[0074] 2,4-Dinitrophenylhydrazine (purity≥98%, Adamas-beta); 5-nitroindole-2-carboxylic acid (purity≥98%, source leaf organisms).

[0075] DMEM high glucose medium (Gibco); high inactivated fetal bovine serum (Gibco); penicillin / streptomycin (Jian...

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PUM

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Abstract

The invention provides a derivatization combined LC-MS method for determining concentration of allantoin in cell matrix. According to the method, glyoxylic acid generated after alkaline hydrolysis andacid hydrolysis of allantoin makes condensation reaction with 2,4-dinitrophenylhydrazine to generate a stable deviation glyoxylic-2,4-dinitrophenylhydrazone. The deviation has relatively high mass spectrum electrospray ionization efficiency, takes 5-nitroindole-2-formic acid as an internal standard substance, and thus, can quantitatively analyze allantoin in a cell sample.

Description

technical field [0001] The invention belongs to the field of analytical chemistry methods, and in particular relates to a method for determining the concentration of allantoin in cells by using derivatization combined with LC-MS technology. [0002] technical background [0003] Aluminum allantoin, alias dihydroxyalantoin aluminum; ureahydrazol; Weichangning; is an antacid and gastric mucosal protective drug. The marketed dosage form is tablet: 100mg, containing 55mg of allantoin and 45mg of aluminum hydroxide. Pharmacological effects of allantoin aluminum: it can accelerate the growth of normal granulation tissue at the injury site and promote the regeneration of mucosal epithelial cells. Indications for allantoin aluminum: digestive tract mucosa protection drug. For gastric ulcer, duodenal ulcer, chronic gastritis. In allantoin aluminum, the structural formula of allantoin is as follows: allantoin is a derivative of urea, Agriculture and light chemical industry. Allant...

Claims

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Application Information

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IPC IPC(8): G01N30/89G01N30/06G01N30/14G01N30/72
CPCG01N30/89G01N30/06G01N30/14G01N30/72G01N2030/062
Inventor 吴凌云於江华王晓霞彭帅
Owner WUXI JIMIN KEXIN SHANHE PHARMA