Determination of Taurine in Biological Samples by High-Performance Liquid Chromatography Using 4-Fluoro-7-Nitrobenzofurazan as a Derivatizing Agent

Objective A highly sensitive and rapid high‐performance liquid chromatography method with pre‐column derivatization with 4‐fluoro‐7‐nitrobenzofurazan was developed for determination of taurine in biological samples,including plasma,brain,and liver.Methods The optimum derivatization reaction temperature was 70℃,and at this temperature the reaction was complete within 3 min.The derivatized taurine was separated using phosphate buffer （0.02 mol/L,pH 6.0）：acetonitrile （84：16,v/v） as the mobile phase at a flow rate of 1.0 mL/min,and a column temperature of 25℃.The taurine derivatives were separated within 20 min （tR：14.5 min） and fluorometrically detected at 530 nm with excitation at 470 nm.Results The intra‐ and the inter‐day coefficients of variation for the method were 5.3% and 7.7%,respectively.The calibration curve was linear from 0.1 μmol/L to 30.0 μmol/L with a correlation coefficient of 0.9995.Conclusion This method can be used to determine the taurine contents in plasma,brain,and liver from normal rats and human plasma.

A novel strategy for screening mutations in the voltage-gated sodium channel gene of Aedes albopictus based on multiplex PCR-mass spectrometry minisequencing technology

Background The current prevention and control strategy forAedes albopictus heavily relies on comprehensive management, such as environmental management and chemical control. However, the wide application of pyrethroids has facilitated the development of insecticide resistance, primarily via mutations in the voltage-gated sodium channel (VGSC) gene. This study aims to develop a novel strategy for detecting mutations in the VGSC gene inAe. albopictus using multiplex PCR-mass spectrometry (MPCR-MS) minisequencing technology.Methods We established a new strategy for detecting mutations in the VGSC gene inAe. albopictus using MPCR-MS minisequencing technology. MPCR amplification and mass probe extension (MPE) were first used, followed by single nucleotide polymorphism (SNP) typing mass spectrometry, which allows the simultaneous detection of multiple mutation sites of the VGSC gene in 96 samples ofAe. albopictus. A total of 70 wild-collectedAe. albopictus were used to evaluate the performance of the method by comparing it with other methods.Results Three target sites (1016, 1532, 1534) in the VGSC gene can be detected simultaneously by double PCR amplification combined with matrix-assisted laser desorption ionization-time-of-flight mass spectrometry, achieving a detection limit of 20 fg/μl. We applied this method to 70 wild-collectedAe. albopictus, and the obtained genotypes were consistent with the routine sequencing results, suggesting the accuracy of our method.Conclusions MPCR-MS minisequencing technology provides a sensitive and high-throughput approach toAe. albopictus VGSC gene mutation screening. Compared with conventional sequencing, this method is economical and time-saving. It is of great value for insecticide resistance surveillance in areas with a high risk of vector-borne disease.