期刊
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
卷 879, 期 32, 页码 3943-3948出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2011.11.008
关键词
Rhodamine 123; Liquid chromatography-tandem mass spectrometry; Rhodamine 6G; Rat plasma
资金
- National Natural Science Foundation of China [81073061]
Rhodamine 123 (R123), as a typical of P-gp substrate, was widely used to quantify P-glycoprotein (P-gp) functional efflux activity in vivo. A new, rapid and sensitive method was developed for quantifying R123 in rat plasma using liquid chromatography-tandem mass spectrometry (LC-MS/MS). R123 and Rhodamine 6G (R6G, the internal standard. IS) were extracted from aliquots of plasma with ethyl acetate and dichloromethane (4:1) as the solvent and 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 -> 4 415.2 was performed. The analysis time was 4 min in positive mode: the calibration curve was linear in the concentration range of 1-200 ng/ml. The lowest limit of quantification (LLOQ) reached 1 ng/ml. The intra and inter-day precision were less than 9.2% for the low quality control (QC) level, and 3.4% for other QC levels, respectively, while the intra and inter-day relative errors ranged between -7.4% and 9.1% for 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 evaluate the functional activity of P-glycoprotein in an absorption experiment in the rat. (C) 2011 Elsevier B.V. All rights reserved.
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