A simple LC-MS/MS method for determination of kynurenine and tryptophan concentrations in human plasma from HIV-infected patients

作者：Huang Yong^*; Louie Alexander; Yang Qiyun; Massenkoff Nicholas; Xu Connie; Hunt Peter W; Gee Winnie

来源：Bioanalysis, 2013, 5(11): 1397-1407.

DOI：10.4155/BIO.13.74

Background: Indoleamine 2,3-dioxygenase, catalyzing tryptophan (Trp) metabolism through the kynurenine (Kyn) metabolic pathway, plays important roles in immune suppression and the CNS. In this article, we report a simple, rapid and specific LC-MS/MS method for accurate determination of Kyn and Trp concentrations in human plasma from HIV-infected patients. Results: The human plasma sample (100 mu l) was mixed with Kyn-d4 and Trp-d5 internal standards and then precipitated with trifluoroacetic acid. The supernatant was directly analyzed by LC-MS/MS. The assay using surrogate matrix calibrators was validated for precision, accuracy, matrix effect, extraction efficiency and stability. Some assay validation issues for endogenous substance bioana-lysis using an LC-MS/MS method are discussed. Conclusion: A simple, specific and reproducible LC-MS/MS method has been developed and validated for measuring Kyn and Trp in human plasma samples.

• 出版日期2013-6