Simultaneous Determination of Seven B Vitamins in Rat Plasma by High-performance Liquid Chromatography-tandem Mass Spectrometry
10.3870/j.issn.1004-0781.2023.12.009
- VernacularTitle:高效液相色谱-串联质谱法同时测定大鼠血浆中7种B族维生素的含量
- Author:
Xiaxia YU
1
;
Yutong ZENG
;
Jiahong YANG
;
Jing WU
;
Yan XU
;
Yin HUANG
Author Information
1. 东南大学附属中大医院药学部,南京 210009
- Keywords:
Vitamin Bs;
High-performance liquid chromatography-tandem mass spectrometry;
Quantitative analysis;
Method validation
- From:
Herald of Medicine
2023;42(12):1796-1801
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a high-performance liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for the simultaneous determination of seven vitamin Bs(VBs)in rat plasma.Methods The plasma samples were pretreated using the protein precipitation method.The chromatographic separation was achieved using an Agilent ZORBAX SB-Aq column(2.1 mm×150 mm,3.5 μm),with a mobile phase composed of 0.01%formic acid(A)and methanol(B)in a gradient elution mode.The flow rate was set at 0.3 mL·min-1 and the column temperature was maintained at 30℃.Mass spectrometric detection was performed using positive ion mode in multiple reaction monitoring.2,4,5,6-Deuteronicotinamide was used as the internal standard.Results Methodological validation results showed that the lowest limits of quantitation(LOQs)for the seven VBs ranged from 7.5 to 300 ng/mL,while the highest LOQs ranged from 1 000 to 20 000 ng·mL-1.Within a certain concentration range,all compounds showed a good linearity(r2>0.992 2).The accuracies ranged from 86.0%to 117.6%,and both intra-and inter-batch precision were less than 20%.Additionally,all analytes demonstrated extraction recoveries greater than 86.1%.No significant matrix effects were observed,and the stability was good.The established validation method was successfully applied to the pharmacokinetics study of single intragastric administration of VB solution in SD rats.Conclusion The analytical method established in this study is simple,rapid,sensitive,and exclusive for the simultaneous determination of VB in real plasma samples.This LC-MS/MS method provides a new analytical tool for further exploring the physiological and pathological effects of VB.
