Sensitive and rapid method for amino acid quantitation in malaria biological samples using AccQ.Tag ultra performance liquid chromatography-electrospray ionization-MS/MS with multiple reaction monitoring.
Sensitive and rapid method for amino acid quantitation in malaria biological samples using AccQ.Tag ultra performance liquid chromatography-electrospray ionization-MS/MS with multiple reaction monitoring.
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DOI：
10.1021/ac901790q
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发表时间：
2010-01-15
影响因子：
7.4
通讯作者：
Shulaev, Vladimir
中科院分区：
文献类型：
--
作者：
Armenta, Jenny M.;Cortes, Diego F.;Pisciotta, John M.;Shuman, Joel L.;Blakeslee, Kenneth;Rasoloson, Dominique;Ogunbiyi, Oluwatosin;Sullivan, David J., Jr.;Shulaev, Vladimir
建立了AccQ·Tag超高效液相色谱-电喷雾串联质谱（AccQ·Tag -UPLC-ESI-MS/MS）法，用于生物样品，特别是恶性疟原虫中氨基酸的快速、重复性好、灵敏度高的定量分析。采用配备光电二极管阵列（PDA）检测器的沃茨Acquity TQD UPLC/MS系统进行氨基酸分离和检测。使用含有酸性、中性和碱性氨基酸的氨基酸标准品混合物开发并验证了该方法。对于MS分析，基于多反应监测的几种选择的AccQ·Tag氨基酸的直接输注分析，实施的最佳锥电压在29-39 V之间变化，而碰撞能量在15-35 V之间变化。使用内部和外部标准化建立校准曲线。通常，在3 × 10−3-25 pmol/μL浓度范围内观察到所有氨基酸的线性响应。对于某些氨基酸，检测的浓度极限低至1.65 fmol。保留时间的变异系数在0.08- 1.08%范围内。通过三次UPLC-MS/MS运行测定的氨基酸定量变异系数平均低于8%。AccQ·Tag-UPLC-ESI-MS/MS方法用于恶性疟原虫和人红细胞样品时显示出良好的技术和生物学重现性。本研究为AccQ·Tag衍生化的UPLC-ESI-MS/MS定量氨基酸提供了有价值的见解。
An AccQ•Tag Ultra performance liquid chromatography-electrospray ionization-tandem mass spectrometry (AccQ•Tag -UPLC-ESI-MS/MS) method for fast, reproducible and sensitive amino acid quantitation in biological samples, particularly, the malaria parasite Plasmodium falciparum is presented. The Waters Acquity TQD UPLC/MS system equipped with photodiode array (PDA) detector was used for amino acid separation and detection. The method was developed and validated using amino acid standard mixtures containing acidic, neutral, and basic amino acids. For MS analysis, the optimum cone voltage implemented, based on direct infusion analysis of a few selected AccQ•Tag amino acids with multiple reaction monitoring, varied from 29-39 V, whereas the collision energy varied from 15-35 V. Calibration curves were built using both internal and external standardization. Typically, a linear response for all amino acids was observed at concentrations ranges of 3 × 10−3-25 pmol/μL. For some amino acids, concentration limits of detection were as low as 1.65 fmol. The coefficients of variation for retention times were within the ranges of 0.08-1.08%. The coefficients of variation for amino acid quantitation, determined from triplicate UPLC-MS/MS runs, were below 8% on the average. The developed AccQ•Tag-UPLC-ESI-MS/MS method revealed good technical and biological reproducibility when applied to P. falciparum and human red blood cells samples. This study should provide a valuable insight into the performance of UPLC-ESI-MS/MS for amino acid quantitation using AccQ•Tag derivatization.
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影响因子：
2.3
作者：
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通讯作者：
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DOI：
10.1016/0378-4347(92)80385-4
发表时间：
1992-09-04
期刊：
JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS
影响因子：
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通讯作者：
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10.1016/s0021-9673(01)94449-8
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期刊：
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作者：
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通讯作者：
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DOI：
10.1016/j.jchromb.2005.12.002
发表时间：
2006-02-02
影响因子：
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作者：
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