Mendez-Alvarez E, Soto-Otero R, Sierra-Paredes G, Aguilar-Veiga E, Galan-Valiente J, Sierra-Marcuño G
University of Santiago, Department of Biochemistry and Molecular Biology, School of Medicine, Santiago de Compostela, Spain.
Biomed Chromatogr. 1991 Mar;5(2):78-82. doi: 10.1002/bmc.1130050207.
A rapid and sensitive high performance liquid chromatographic method is described for the simultaneous determination of benzylpenicillin, ampicillin, phenoxymethylpenicillin, cloxacillin, dicloxacillin and nafcillin in small samples of human serum. The chromatographic system involves the use of a Spherisorb ODS reversed phase column and a gradient elution with 1 mM ammonium acetate buffer/acetonitrile (from 90:10 to 75:25 in 15 min). Detection and quantification are monitored by UV absorption at 208 nm. The compounds are extracted with dichloromethane, using tetrabutylammonium hydrogen sulfate neutralized with sodium hydroxide and buffered with borate as an ion pairing reagent; beta-hydroxyethyltheophylline is added as an internal standard. Our results show that the method is accurate and reproducible, allowing quantification of serum levels of assayed penicillins (0.5-50 micrograms/mL) without interference from other drugs commonly used in therapy. Recoveries were generally greater than 79.4%.
本文描述了一种快速灵敏的高效液相色谱法，用于同时测定人血清小样本中的苄青霉素、氨苄青霉素、苯氧甲基青霉素、氯唑西林、双氯西林和萘夫西林。色谱系统包括使用Spherisorb ODS反相柱和用1 mM醋酸铵缓冲液/乙腈进行梯度洗脱（在15分钟内从90:10变为75:25）。通过在208 nm处的紫外吸收进行检测和定量。化合物用二氯甲烷萃取，使用用氢氧化钠中和并用硼酸盐缓冲的硫酸氢四丁铵作为离子对试剂；加入β-羟乙茶碱作为内标。我们的结果表明，该方法准确且可重复，能够对所测青霉素的血清水平（0.5 - 50微克/毫升）进行定量，而不受治疗中常用的其他药物干扰。回收率一般大于79.4%。
