Smyrniotakis C G, Archontaki Helen A
Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Athens 15771, Greece.
J Pharm Biomed Anal. 2007 Jan 17;43(2):506-14. doi: 10.1016/j.jpba.2006.07.048. Epub 2006 Sep 20.
Simultaneous determination of oxytetracycline, 4-epioxytetracycline, alpha-apooxytetracycline, tetracycline and beta-apooxytetracycline on C(18) columns has been accomplished using a high performance liquid chromatographic method with UV detection. Separation was achieved on a Hypersil BDS RP-C(18) column (250 mm x 4.6 mm) and on a Waters C(18) Symmetry column (150 mm x 3.9mm), 5 microm particle size each. These columns were equilibrated with mobile phases consisted of methanol-acetonitrile-0.1M phosphate buffer pH 8.0 (12.5:12.5:75, v/v/v) and (15:15:70, v/v/v), respectively. The flow rate was 1.0 ml/min and the total elution time was 15 and 5 min, respectively. Both methods were applied to oxytetracycline raw material, human and veterinary formulations, where the excipients did not interfere. External standard calibration curves were linear for 4-epioxytetracycline, oxytetracycline, alpha-apooxytetracycline, tetracycline and beta-apooxytetracycline in the concentration range of 0.27-200 microM, 0.05-200 microM, 0.03-200 microM, 0.35-200 microM and 0.20-200 microM on column A and 0.08-200 microM, 0.15-200 microM, 0.09-200 microM, 0.25-200 microM and 0.47-200 microM on column B, respectively. Day-to-day relative standard deviation of the determination for every component was less than 3%. Concerning the first column, limits of detection and quantification of the above compounds were in the concentration ranges of 10-106 nM and 30-352 nM, respectively, whereas on the second column these ranges became 27-144 nM and 81-475 nM, respectively. Recovery of the separated compounds was 95-105%.
采用带紫外检测的高效液相色谱法，在C(18)柱上同时测定土霉素、4-表土霉素、α-脱水土霉素、四环素和β-脱水土霉素。在Hypersil BDS RP-C(18)柱（250 mm×4.6 mm）和Waters C(18) Symmetry柱（150 mm×3.9mm）上实现分离，两柱粒径均为5微米。这些柱子分别用由甲醇-乙腈-0.1M pH 8.0磷酸盐缓冲液（12.5:12.5:75，v/v/v）和（15:15:70，v/v/v）组成的流动相平衡。流速为1.0 ml/min，总洗脱时间分别为15分钟和5分钟。两种方法均应用于土霉素原料药、人用和兽用制剂，其中辅料不产生干扰。4-表土霉素、土霉素、α-脱水土霉素、四环素和β-脱水土霉素的外标校准曲线在浓度范围0.27 - 200 microM、0.05 - 200 microM、0.03 - 200 microM、0.35 - 200 microM和0.20 - 200 microM（柱A）以及0.08 - 200 microM、0.15 - 200 microM、0.09 - 200 microM、0.25 - 200 microM和0.47 - 200 microM（柱B）内呈线性。各组分测定的日间相对标准偏差均小于3%。对于第一根柱子，上述化合物的检测限和定量限分别在10 - 106 nM和30 - 352 nM浓度范围内，而在第二根柱子上，这些范围分别变为27 - 144 nM和81 - 475 nM。分离化合物的回收率为95 - 105%。
