A simple LC-MS/MS method for determination of kynurenine and tryptophan concentrations in human plasma from HIV-infected patients.
A simple LC-MS/MS method for determination of kynurenine and tryptophan concentrations in human plasma from HIV-infected patients.
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作者：
Huang Y;Louie A;Yang Q;Massenkoff N;Xu C;Hunt PW;Gee W
吲哚胺2，3-双加氧酶通过犬尿氨酸（Kyn）代谢途径催化色氨酸（Trp）代谢，在免疫抑制和CNS中起重要作用。在本文中，我们报告了一种简单，快速和特异性的LC-MS/MS方法，用于准确测定HIV感染者血浆中Kyn和Trp的浓度。将人血浆样品（100 μl）与Kyn-d4和Trp-d5内标物混合，然后用三氟乙酸沉淀。通过LC-MS/MS直接分析上清液。对使用替代基质校准品的测定法的精密度、准确度、基质效应、提取效率和稳定性进行了验证。本文讨论了LC-MS/MS法用于内源性物质生物分析的一些检测验证问题。已开发并验证了一种简单、特异性和重现性良好的LC-MS/MS方法，用于测定人血浆样品中的Kyn和Trp。
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. The human plasma sample (100 μl) 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 bioanalysis using an LC–MS/MS method are discussed. A simple, specific and reproducible LC–MS/MS method has been developed and validated for measuring Kyn and Trp in human plasma samples.
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DOI：
10.1016/j.jchromb.2012.04.030
发表时间：
2012-06-01
影响因子：
3
作者：
Moeller, Marisa;Du Preez, Jan L.;Harvey, Brian H.
通讯作者：
Harvey, Brian H.
影响因子：
2
作者：
Jemal, M;Schuster, A;Whigan, DB
通讯作者：
Whigan, DB
DOI：
10.1016/s1044-0305(96)00072-4
发表时间：
1996-11-01
影响因子：
3.2
作者：
Buhrman, DL;Price, PI;Rudewicz, PJ
通讯作者：
Rudewicz, PJ
影响因子：
17.1
作者：
Favre D;Mold J;Hunt PW;Kanwar B;Loke P;Seu L;Barbour JD;Lowe MM;Jayawardene A;Aweeka F;Huang Y;Douek DC;Brenchley JM;Martin JN;Hecht FM;Deeks SG;McCune JM
通讯作者：
McCune JM
影响因子：
7.4
作者：
Li, WL;Cohen, LH
通讯作者：
Cohen, LH