高效液相色谱法测定感染敏感和耐药恶性疟原虫疟疾患者血浆中的周效磺胺和N - 乙酰周效磺胺。
High-performance liquid chromatographic assay for the determination of sulfadoxine and N-acetyl sulfadoxine in plasma from patients infected with sensitive and resistant Plasmodium falciparum malaria.
作者信息
Dua Virendra K, Gupta N C, Sethi Prerana, Edwards G, Dash A P
机构信息
National Institute of Malaria Research, Field Unit, Sector-III, BHEL, Hardwar 249403, India.
出版信息
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Dec 15;860(2):160-5. doi: 10.1016/j.jchromb.2007.10.016. Epub 2007 Oct 22.
A reversed-phase high-performance liquid chromatographic method using a mobile phase of acetonitrile-methanol-trifluoroacetic acid-water (16.1:7.2:0.1:76.6, v/v/v/v) at a flow rate of 1.0 ml min(-1) on a LiChrospher RP-18 column with UV (254 nm) detection has been developed for the separation of sulfadoxine and its metabolite N-acetyl sulfadoxine in plasma. No interferences due to endogenous compounds or common antimalarial drugs were noticed. The limit of detection for sulfadoxine and N-acetyl sulfadoxine was 0.01 microg ml(-1) with a signal-to-noise ratio of 5:1 while the limit of quantification was 2.5 microg ml(-1). Intra-day mean relative standard deviations (RSD's) for sulfadoxine and N-acetyl sulfadoxine were 2.6 and 2.8%, respectively, while mean inter-day RSD's for sulfadoxine and N-acetyl sulfadoxine were 2.4 and 2.8%, respectively. Extraction recoveries averaged 90.6% for sulfadoxine and 86.9% for N-acetyl sulfadoxine. The method was applied for the assay of sulfadoxine and its metabolite N-acetyl sulfadoxine in plasma from Plasmodium falciparum malaria patients. Mean plasma sulfadoxine concentrations on day 2 (51 h) from samples collected from sensitive and resistant P. falciparum patients treated with three tablets of Fansidar were 62.8 and 60.5 microg ml(-1), respectively. Mean ratio of N-acetyl sulfadoxine to sulfadoxine was 9.1% for responders and 13.9% for non-responders which revealed that higher amounts of the metabolite N-acetyl sulfadoxine were present in non-responders. The method described should find an application in the therapeutic monitoring of malaria patients.
已开发出一种反相高效液相色谱法，该方法使用乙腈 - 甲醇 - 三氟乙酸 - 水（16.1:7.2:0.1:76.6，v/v/v/v）的流动相，流速为1.0 ml min⁻¹，在LiChrospher RP - 18柱上进行分离，并采用紫外（254 nm）检测，用于分离血浆中的磺胺多辛及其代谢物N - 乙酰磺胺多辛。未发现内源性化合物或常见抗疟药物的干扰。磺胺多辛和N - 乙酰磺胺多辛的检测限为0.01 μg ml⁻¹，信噪比为5:1，定量限为2.5 μg ml⁻¹。磺胺多辛和N - 乙酰磺胺多辛的日内平均相对标准偏差（RSD）分别为2.6%和2.8%，而日间平均RSD分别为2.4%和2.8%。磺胺多辛的提取回收率平均为90.6%，N - 乙酰磺胺多辛的提取回收率平均为86.9%。该方法用于测定恶性疟原虫疟疾患者血浆中的磺胺多辛及其代谢物N - 乙酰磺胺多辛。从服用三片 Fansidar 治疗的敏感和耐药恶性疟患者收集的样本中，第2天（51小时）血浆磺胺多辛的平均浓度分别为62.8和60.5 μg ml⁻¹。反应者中N - 乙酰磺胺多辛与磺胺多辛的平均比值为9.1%，无反应者为13.9%，这表明无反应者中存在较高量的代谢物N - 乙酰磺胺多辛。所描述的方法应可应用于疟疾患者的治疗监测。
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