Maher Hadir M, Youssef Rasha M, Khalil Riad H, El-Bahr Sabry M
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, University of Alexandria, El-Messalah, Alexandria 21521, Egypt.
J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Dec 15;876(2):175-81. doi: 10.1016/j.jchromb.2008.10.033. Epub 2008 Nov 1.
An efficient multiresidue method for the simultaneous determination of metronidazole (MET) and spiramycin (SPY) in tilapia fish muscle, based on high performance liquid chromatography with UV detection (HPLC-UV), has been developed. The drugs were extracted with 0.2% orthophosphoric acid-methanol (6:4), and the extracts were cleaned up on a solid phase extraction cartridge, C18 Sep-Pak light column. The LC separation was performed on a RP stainless-steel C-18 analytical column (150 mm x 4.6 mm, 5 microm) with a gradient elution system of 0.05 M phosphate buffer adjusted to pH 2.4-acetonitrile as the mobile phase at the flow rate of 1.0 ml min(-1). A wavelength programming was applied for the UV detection of the analytes. The method not only enabled the determination of the parent drugs, MET and SPY, but also permitted the determination of their metabolites, hydroxymetronidazole (HMET) and neospiramycin (NSPY). The calibration graphs for each drug were rectilinear in the range of 0.005-1.000 microg g(-1) for MET and HMET and 0.025-1.000 microg g(-1) for SPY and NSPY. With this method, the cited drugs with their metabolites were determined in fortified fish muscle tissues at levels of 0.025, 0.1 and 1.0 microg g(-1) with good accuracy and precision. LOD and LOQ obtained for each drug were as follows: 0.002 and 0.005 microg g(-1) for MET and HMET and 0.005 and 0.025 microg g(-1) for SPY and NSPY. Utilization of the method to successfully analyze tilapia fish muscle samples incurred with MET and SPY was described.
基于高效液相色谱-紫外检测法（HPLC-UV），开发了一种同时测定罗非鱼肌肉中甲硝唑（MET）和螺旋霉素（SPY）的高效多残留方法。药物用0.2%正磷酸-甲醇（6:4）提取，提取物在固相萃取柱C18 Sep-Pak轻柱上进行净化。液相色谱分离在RP不锈钢C-18分析柱（150 mm×4.6 mm，5μm）上进行，以0.05 M磷酸盐缓冲液（pH值调至2.4）-乙腈为流动相，采用梯度洗脱系统，流速为1.0 ml min⁻¹。对分析物进行紫外检测时采用波长程序设定。该方法不仅能够测定母体药物MET和SPY，还能测定它们的代谢产物羟基甲硝唑（HMET）和新螺旋霉素（NSPY）。每种药物的校准曲线在0.005 - 1.000 μg g⁻¹范围内呈线性，其中MET和HMET为该范围，SPY和NSPY为0.025 - 1.000 μg g⁻¹。用该方法在添加了目标药物的罗非鱼肌肉组织中测定目标药物及其代谢产物，添加水平为0.025、0.1和1.0 μg g⁻¹，具有良好的准确度和精密度。每种药物的检测限和定量限如下：MET和HMET分别为0.002和0.005 μg g⁻¹，SPY和NSPY分别为0.005和0.025 μg g⁻¹。描述了该方法成功分析受MET和SPY污染的罗非鱼肌肉样品的应用情况。
