El-Rabbat Nawal, Askal Hassan F, Khashaba Pakinaz Y, Attia Noha N
Assiut University, Faculty of Pharmacy, Department of Pharmaceutical Analytical Chemistry, Assuit, Egypt.
J AOAC Int. 2006 Sep-Oct;89(5):1276-87.
This paper describes a simple spectrofluorometric method for the analysis of 4 macrolide antibiotics. The method is based on the condensation of 10% (w/v) malonic acid and acetic acid anhydride under the catalytic effect of tertiary amine groups of the studied macrolides. The relative fluorescence intensity of the condensation product was measured at 397/452 nm (excitation/emission) for azithromycin dihydrate and at 392/445 nm (for clarithromycin, erythromycin ethylsuccinate, and roxithromycin. All variables affecting the reaction conditions were studied. The effects of potential interference due to common excipients, such as starch, lactose, sucrose, glucose, gum acacia, and magnesium stearate, as well as trimethoprim and sulfisoxazole acetyl formulated in primomycin capsules and pediazole oral suspension, respectively, were studied. A validation study for the proposed method was carried out according to U.S. Pharmacopeia 2002. The linearity ranges were 3-80 ng/mL for all of the cited macrolides. The limit of detection range was 0.74-1.20 ng/mL, while the limit of quantitation range was 2.47-4.02 ng/mL. The method was applied for the assay of the studied macrolides in pure pharmaceutical formulations and in spiked biological fluids. Results were compared with those obtained from the reported method, where calculated t- and F-values indicated high accuracy and good precision for the proposed method.
本文描述了一种用于分析4种大环内酯类抗生素的简单荧光分光光度法。该方法基于在被研究大环内酯类的叔胺基团催化作用下，10%（w/v）丙二酸与乙酸酐的缩合反应。对于二水阿奇霉素，在397/452 nm（激发/发射）处测量缩合产物的相对荧光强度；对于克拉霉素、琥乙红霉素和罗红霉素，则在392/445 nm处测量。研究了所有影响反应条件的变量。研究了常见辅料（如淀粉、乳糖、蔗糖、葡萄糖、阿拉伯胶和硬脂酸镁）以及分别在利福平胶囊和复方磺胺异恶唑口服混悬液中配制的甲氧苄啶和磺胺异恶唑乙酰的潜在干扰影响。根据《美国药典》2002年版对所提出的方法进行了验证研究。所有上述大环内酯类的线性范围为3 - 80 ng/mL。检测限范围为0.74 - 1.20 ng/mL，而定量限范围为2.47 - 4.02 ng/mL。该方法应用于纯药物制剂和加标生物流体中所研究大环内酯类的含量测定。将结果与从报道方法获得的结果进行比较，计算得到的t值和F值表明所提出的方法具有高准确性和良好的精密度。
