Dua Virendra K, Gupta N C, Sethi Prerana, Edwards G, Dash A P
National Institute of Malaria Research, Field Unit, Sector-III, BHEL, Hardwar 249403, India.
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。
