High-performance liquid chromatographic assay for the quantitation of L-arginine in human plasma
High-performance liquid chromatographic assay for the quantitation of L-arginine in human plasma
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DOI：
10.1021/ac960613n
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发表时间：
1996-10-01
影响因子：
7.4
通讯作者：
Blaschke, TF
中科院分区：
文献类型：
--
作者：
Gopalakrishnan, V;Burton, PJ;Blaschke, TF
L-精氨酸通过一氧化氮合酶代谢为一氧化氮，一氧化氮产生的异常与一些涉及脉管系统的疾病的发病机制有关，因此，人们对L-精氨酸的药理剂量对心血管和肾脏疾病患者的影响产生了兴趣。为了研究外源剂量的处置，开发了一种 HPLC 方法来分析血浆样品中的 L-精氨酸。该测定涉及用萘二甲醛和氰化物对精氨酸进行柱前衍生化，然后进行 HPLC 和 UV 检测。只需对血浆样品进行简单的脱蛋白处理，衍生化的精氨酸稳定（20小时内降解小于5%），有利于自动进样器中的批量样品处理和分析。校准曲线是在林格氏乳酸盐溶液中而不是血浆中生成的，以校正内源性血浆 L-精氨酸。与林格氏溶液 (n = 4) 相比，血浆中的回收率为 103%。平均日内检测精度 (n = 6)（以变异系数表示）为 3.4%。批间精密度 (n = 6) 为 7%。该测定法适用于对来自正常受试者的血浆样本中的 L-精氨酸进行定量，该正常受试者已接受单次口服（10 g）和单次静脉注射（30 g）的性感 L-精氨酸。
L-Arginine is metabolized to nitric oxide by nitric oxide synthase, and abnormalities in nitric oxide production have been implicated in the pathogenesis of some diseases involving the vasculature, Thus, there has been interest in the effects of pharmacologic doses of L-arginine in patients with cardiovascular and renal diseases. To study the disposition of exogenous doses, an HPLC method was developed to analyze plasma samples for L-arginine, The assay involves precolumn derivatization of arginine with naphthalenedicarboxaldehyde and cyanide followed by HPLC with UV detection. Only a simple deproteinization of the plasma samples was required, The derivatized arginine was stable (less than 5% degradation in 20 h), facilitating batch sample processing and analysis in an autosampler. Calibration curves were generated in Ringer's lactate solution instead of plasma to correct for endogenous plasma L-arginine. Recovery in plasma, compared to Ringer's solution (n = 4), was 103%. Mean intraday assay precision (n = 6), expressed as coefficient of variation, was 3.4%. Interassay precision (n = 6) was 7%. The assay was applied for the quantitation of L-arginine in plasma samples from a normal subject who had been given a single oral (10 g) and a single intravenous dose (30 g) of erogenous L-arginine.