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[高压液相色谱法测定血浆草酸]

[Determination of plasma oxalic acid by high-pressure liquid chromatography].

作者信息

Jerez E

出版信息

Rev Esp Fisiol. 1986 Dec;42(4):441-7.

PMID:3563073
Abstract

A new specific and sensitive method for determination of oxalic acid in plasma by High Performance Liquid Chromatography (HPLC) is described. The plasma sample is deproteinized by ultrafiltration. The oxalic acid in the ultrafiltrate is purified by precipitation with CaCl2, new dilution of calcium oxalate precipitate, oxalic acid extraction with diethyl-ether and total dryness of the sample. The losses of oxalic acid during this process are evaluated by the addition of oxalic acid (U-14C) before the precipitation step. The dried samples are redissolved in mobile phase (o-H3PO4, 0.05 M) and injected into a HPLC chromatograph, with reversed phase column (Lichrosorb RP-8, Merck). Oxalate peak is detected spectrophotometrically at 220 nm with a retention time of 3.20 minutes. The method shows a mean recovery value of 82.11, with an intra-run and between-run CV values of 2.54 and 6.95 respectively. The oxalic acid measured in plasma by this method is 291 +/- 89 micrograms/100 ml plasma ultrafiltrate, in 16 normal subjects.

摘要

本文描述了一种通过高效液相色谱法(HPLC)测定血浆中草酸的新的特异性和灵敏性方法。血浆样品通过超滤进行脱蛋白处理。超滤液中的草酸通过用氯化钙沉淀、新稀释草酸钙沉淀、用乙醚萃取草酸以及样品完全干燥来进行纯化。在此过程中草酸的损失通过在沉淀步骤之前加入草酸(U-14C)来评估。干燥后的样品重新溶解于流动相(邻磷酸,0.05M)中,并注入配有反相柱(Lichrosorb RP-8,默克公司)的HPLC色谱仪中。草酸盐峰在220nm处通过分光光度法检测,保留时间为3.20分钟。该方法的平均回收率为82.11,批内和批间CV值分别为2.54和6.95。在16名正常受试者中,用该方法测得的血浆中超滤液草酸含量为291±89微克/100毫升血浆。

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