Using liquid chromatography tandem mass spectrometry to detect rhodamine123 in rat plasma

作     者：Ya-Bin Sun~a,Zhong-Yuan Xu~a,Guo-Feng Li~b a GCP Office,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China b Department of Pharmacy,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China 

会议名称：《第十三次全国临床药理学学术大会》

会议日期：2012年

学科分类：1006[医学-中西医结合] 10[医学] 100602[医学-中西医结合临床] 

基　　金：supported by a grant from the National Natural Science Foundations of China(N08 1073061) the Foundation for Sci & Tech Research Project of Guangdong Province,China(2010B030700014) the Foundation for Sci & Tech Research Project of Baiyun branch,Guangzhou,China(2009-sz-36) 

关 键 词：Rhodamine 123 Liquid chromatography tandem mass spectrometry Rhodamine 6G Rat plasma 

摘      要：A new,rapid and sensitive method was developed for the quantifying of rhodamine 123(R123) in rat plasma using liquid chromatography tandem mass spectrometry(LC-MS/MS).Rhodamine 6G(R6G) was used as the internal standard(IS).R123 and IS were extracted from aliquots of plasma with ethyl acetate and dichloromethane (4:1) as the solvent.Chromatographic separation was performed using a Zorbax Eclipse Plus C18 column.The mobile phase was composed of A:ammonium formate-formic acid buffer containing 5 mM ammonium formate and 0.1%formic acid and B:methanol(A:B,5:95,v/v).To quantify R123 and IS respectively,multiple reaction monitoring(MRM) transition of m/z 345.2→285.2 and m/z 443.3→415.2 were performed.The analysis lasted for 4 min,in the positive mode,the results of which were presented as a linear calibration curve over the concentration range of 1-200 ng/ml and sampling volume of 2μl.The lowest limit of quantification(LLOQ) reached 1 ng/ml.The intra and inter-day precision were 6.7%and 9.2%for the low quality control(QC) sample (2 ng/ml),respectively.Intra and inter-day precision of the medium QC sample(100 ng/ml) were both less than 3.4%.Intra and inter-day precision of high QC sample(150 ng/ml) were both less than 2.1%,while the intra and inter-day relative errors ranged between -7.4%and 9.1%for the above three QC concentration levels.The LC-MS/MS method proved to be simple,accurate,reliable and with a shorter running time and has been successfully applied to an absorption experiment in the rat.