Method for measuring aescin A, B, C and D in human plasma through utilization of LC-MSMS method and application thereof

Abstract

The invention discloses a method for measuring aescin A, B, C and D in human plasma through utilization of an LC-MSMS method. The method comprises the following steps of carrying out acidizing; carrying out high speed centrifugation; carrying out drying through utilization of a termovap sample concentrator; and carrying out redissolving and swirling. According to the method, a sample is processedthrough precipitation of protein by methanol, and after nitrogen drying and redissolving are carried out, the sample is injected. Compared with solid phase extraction, the method has the advantages that a recovery rate is stable, processing time is greatly reduced, cost is clearly reduced, four compositions are measured, quantitation lower limits are clearly separated, the quantitation lower limits of aescin A, B, C and D are 20pg / mL, 20pg / mL, 40pg / mL and 40pg / mL, and an oral pharmacokinetic sample measurement requirement is satisfied. Plasma is added to acidizing stabilizer. ISR analysis qualification rates of the four compositions are more than 95%. The method is applied to aescin oral preparation clinical pharmacokinetic study. A pharmacokinetic behavior can be drawn completely.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for measuring aescin A, B, C and D in human plasma by LC-MSMS and an application thereof. Background technique [0002] The molecular structure of aescin is composed of saponins and sugars connected by glycosidic bonds. The molecular polarity is relatively large, and it is difficult to dissolve in low-polar organic solvents. It is difficult to extract it from biological samples by conventional liquid-liquid extraction methods. Therefore, most of the samples are processed by methanol precipitation or solid phase extraction. There is a lack of chromophore in the molecular structure of the components of aescin, and conventional analysis methods such as HPLC-UV cannot provide sufficient sensitivity, so the analysis of its concentration in vivo has become a bottleneck limiting the study of the pharmacokinetics of aescin. In 1991, Kunz et al. used immunoassay (RIA) for th...

AI Technical Summary

Problems solved by technology

There is a lack of chromophores in the molecular structure of the components of aescin, and conventional analysis methods such as HPLC-UV cannot provide sufficient sensitivity, so the concentration analysis in vivo has become a bottleneck limiting the pharmacokinetics of aescin

[0003] Solid-phase extraction is used for biological samples, the steps are cumbersome, and the cost of sample processing is high, which seriously affects the analysis efficiency

The four components of aescin A, B, C, and D in the biological matrix were only partially determined or the four components did not reach the baseline separation. The lower limit of quantification of the current detection method is too high to meet the requirements for the analysis of biological samples after oral administration. Aescin Saponins A and C, B and D are interconverted, no stabilization measures have been taken

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