Wang Li-Feng, Peng Jing-Dong, Liu Li-Min
School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, China.
Anal Chim Acta. 2008 Dec 7;630(1):101-6. doi: 10.1016/j.aca.2008.09.066. Epub 2008 Oct 14.
A new reversed-phase high performance liquid chromatography with resonance Rayleigh scattering detection (HPLC-RRS) was developed for simultaneous separation and determination of four tetracycline antibiotics (TCs). A good chromatographic separation among the compounds was achieved using a Synergi Fusion-RP column (150 mm x 4.6 mm; 4 microm) and a mobile phase consisting of methanol-acetonitrile-oxalic acid (5mM) at the flow rate of 0.8 mLmin(-1). Column temperature was 30 degrees C. The RRS signal was detected at lambda(ex)=lambda(em)=370 nm. The recoveries of sample added standard ranged from 95.3% to 103.5%, and the relative standard deviation was below 2.79%. A detection limit of 2.12-5.12 microgmL(-1) was reached and a linear range was found between peak height and concentration in the range of 10.36-518.0 microgmL(-1) for oxytetracycline (OTC), 12.11-605.5 microgmL(-1) for tetracycline (TC), 11.79-589.5 microgmL(-1) for chlortetracycline (CTC) and 10.32-516.0 microgmL(-1) for doxycycline (DC). The linear regression coefficients were all above 0.999. The method has been applied successfully to the determination of OTC, TC, CTC, DC in pharmaceutical formulations and in honey. The method was simple, rapid and showed a better linear relation and high repeatability.
建立了一种用于同时分离和测定四种四环素类抗生素（TCs）的新型反相高效液相色谱-共振瑞利散射检测法（HPLC-RRS）。使用Synergi Fusion-RP柱（150 mm×4.6 mm；4μm）和由甲醇-乙腈-草酸（5 mM）组成的流动相，流速为0.8 mLmin⁻¹，实现了各化合物之间良好的色谱分离。柱温为30℃。在λ(ex)=λ(em)=370 nm处检测RRS信号。加标样品的回收率在95.3%至103.5%之间，相对标准偏差低于2.79%。土霉素（OTC）的检出限为2.12 - 5.12μgmL⁻¹，土霉素（OTC）在10.36 - 518.0μgmL⁻¹范围内、四环素（TC）在12.11 - 605.5μgmL⁻¹范围内、金霉素（CTC）在11.79 - 589.5μgmL⁻¹范围内、强力霉素（DC）在10.32 - 516.0μgmL⁻¹范围内，峰高与浓度之间呈现线性关系。线性回归系数均大于0.999。该方法已成功应用于药物制剂和蜂蜜中土霉素（OTC）、四环素（TC）、金霉素（CTC）、强力霉素（DC）的测定。该方法简便、快速，具有较好的线性关系和高重复性。
