Szoke A, Hayton W L, Schultz I R
Ohio State University, College of Pharmacy, Columbus 43210, USA.
J Pharm Biomed Anal. 1997 Sep;16(1):69-75. doi: 10.1016/s0731-7085(97)00014-9.
Methods for extraction and gradient HPLC quantification were developed for benzocaine (BZ) and three of its metabolites to be used in conjunction with a reverse isotope technique. The metabolites were p-aminobenzoic acid (PABA), acetyl-p-aminobenzoic acid (AcPABA) and acetylbenzocaine (AcBZ). The matrixes studied were white muscle, red muscle, skin, liver, trunk kidney, head kidney, plasma and the bile of channel catfish. Analytes were validated for each of the compounds at 25 and 100 nmol per sample in the various tissues and fluids. The intraday variability (R.S.D.) was less than 13% in all tissues and fluids except for BZ in the liver. Recoveries varied from matrix to matrix for each analyte. The highest recoveries were obtained from plasma which ranged from 82.8-99.8% depending on the concentration. The average recovery of the compounds from tissues was between 50 and 78%, except for liver where the recovery of PABA and BZ was below 30%. Detection was by UV absorbance at 286 nm and the linear range was 2.5-15 nmol 100 ml-1 for all analytes. The method was selective; no interference peaks coeluted with the analytes.
开发了用于苯佐卡因（BZ）及其三种代谢物的提取和梯度高效液相色谱定量方法，以与反向同位素技术结合使用。这些代谢物是对氨基苯甲酸（PABA）、乙酰对氨基苯甲酸（AcPABA）和乙酰苯佐卡因（AcBZ）。所研究的基质包括斑点叉尾鮰的白肌、红肌、皮肤、肝脏、躯干肾、头肾、血浆和胆汁。在各种组织和液体中，对每种化合物在每个样品25和100 nmol的水平进行了分析物验证。除肝脏中的BZ外，所有组织和液体中的日内变异性（相对标准偏差）均小于13%。每种分析物在不同基质中的回收率各不相同。血浆中的回收率最高，根据浓度不同，范围为82.8 - 99.8%。除肝脏中PABA和BZ的回收率低于30%外，化合物在组织中的平均回收率在50%至78%之间。通过286 nm处的紫外吸光度进行检测，所有分析物的线性范围为2.5 - 15 nmol 100 ml-1。该方法具有选择性；没有干扰峰与分析物共洗脱。
