基于气相色谱质谱联用法对金黄色葡萄球菌中6种有机酸的测定  

Determination of six organic acids in Staphylococcus aureus by gas chromatography mass spectrometry

作　　者：叶霜 张斌[1] 胡小爱 杨晨[1] 鲁兰[1] 龙姗姗 刘琦珺 颜军[1] Ye Shuang;Zhang Bin;Hu Xiao-ai;Yang Chen;Lu Lan;Long Shan-shan;Liu Qi-jun;Yan Jun(Antibiotics Research and Re-evaluation Key Laboratory of Sichuan Province,Sichuan Industrial Institute of Antibiotics,School of pharmacy,Chengdu University,Chengdu 610106;School of Food and Bioengineering,Chengdu University,Chengdu 610106;Gansu Jingning Hospital of Traditional Chinese Medicine,Jingning 743400)

机构地区：[1]抗生素研究与再评价四川省重点实验室,四川抗菌素工业研究所,药学院,成都大学,成都610106 [2]成都大学食品与生物工程学院,成都610106 [3]甘肃省静宁县中医医院,静宁743400

出　　处：《中国抗生素杂志》2022年第12期1267-1273,共7页Chinese Journal of Antibiotics

基　　金：国家自然科学基金(No.81803812);四川省国际科技创新合作项目(No.2019YFH0054和No.2020YFH0205)。

摘　　要：目的采用气相色谱质谱联用法(GC-MS)建立临床耐药金黄色葡萄球菌中琥珀酸、延胡索酸、苹果酸、草酰乙酸、α-酮戊二酸及柠檬酸的测定方法。方法6种有机酸由硅烷化试剂N,O-双(三甲基硅烷基)三氟乙酰胺(含1%三甲基氯硅烷)(BSTFA-TMCS)衍生化后,采用选择离子扫描模式(SIR)以外标法进行定量测定。结果6种有机酸在0.31~300μg/mL范围内线性良好,相关系数大于0.99,回收率在83.02%~109.28%之间。同时,利用该方法对乳酸左氧氟沙星处理后的金黄色葡萄球菌胞内6种有机酸含量进行测定,与未经乳酸左氧氟沙星处理的SA相比6种有机酸代谢含量均表达下调。结论该方法受基质影响小,重现性好,适合于复杂生物基质样本中有机酸含量的测定,同时发现乳酸左氧氟沙星是通过柠檬酸循环途径抑制金黄色葡萄球菌生长。Objective A method for the determination of succinic acid,fumaric acid,malic acid,oxaloacetic acid,α-ketoglutaric acid,and citric acid in clinical drug-resistant Staphylococcus aureus was established by gas chromatography-mass spectrometry(GC-MS).Methods The six organic acids were derivatized with N,Obis(trimethylsilane)trifluoroacetamide(containing 1%trimethylchlorosilane)(BSTFA-TMCS)as the silanization reagent and quantitated using external standard method under SIR model.Results The six organic acids showed good linearity in the range of 0.31~300μg/mL,the correlation coefficient was greater than 0.99,and the recoveries were in the range of 83.02%~109.28%.At the same time,this method was used to determine the contents of the six organic acids in the cells of Staphylococcus aureus(SA)treated with levofloxacin lactate.Compared with SA treated without levofloxacin lactate,the metabolic levels of the six organic acids were down-regulated.Conclusion This method is suitable for the determination of organic acids in complex biological matrix samples,and it is found that levofloxacin lactate inhibits the growth of Staphylococcus aureus through the citric acid cycle pathway.

关 键 词：气相色谱质谱联用法 衍生化反应 柠檬酸循环 有机酸 

分 类 号：R917[医药卫生—药物分析学]