An Improved High‐Performance Liquid Chromatographic Method for the Simultaneous Measurement of Halofantrine and Desbutylhalofantrne in Human Serum
作者:
Duangsuda Keeratithakul,
Paktiya Teja-Isavadharm,
G. Shanks,
H. Webster,
Michael Edstein,
期刊:
Therapeutic Drug Monitoring
(OVID Available online 1991)
卷期:
Volume 13,
issue 1
页码: 64-68
ISSN:0163-4356
年代: 1991
出版商: OVID
关键词: Halofantrine;Desbutylhalofantrine;Malaria;High-performance liquid chromatography
数据来源: OVID
摘要:
A rapid, accurate, and sensitive high-performance liquid chromatographic (HPLC) method, with ultraviolet detection, for simultaneous measurement of halofantrine (HAL) and desbutylhalofantrine (BHAL) in human serum is described. Sample preparation involved protein precipitation, followed by a solid-phase clean-up and a liquid-liquid extraction. Chromatographic separation was carried out on an Ultrasphere C8 column (25 cm x 4.6 mm I.D., 5 μm particle size) using a mobile phase of acetonitrile:water, 75:25, (vol/vol), with 0.2% (w/vol) sodium dodecyl sulfate and 0.2% (vol/vol) glacial acetic acid. The serum sample volume used was 0.5 ml, with concentrations normalized to 1 ml. Retention times for BHAL, HAL, and the internal standard were 5.5, 8.3, and 11.5 min, respectively, with a total run time of 13.5 min. The average extraction recovery for HAL was 85.6% and 100.5% for BHAL. Inter- and intra-assay precisions of HAL and BHAL were $7.5%, with an accuracy of $10.8% over three concentrations (0.02, 0.5, 2.0 μg/ml). Detection limits of HAL and BHAL were 5 and 3 ng/ml, respectively. The new HPLC method resulted in cleaner chromatograms and a 1.7-fold faster run time than previous HPLC methods. Application of the method with clinical specimens was demonstrated.
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