Emami Jaber, Rezazadeh Mahboubeh
Department of Pharmaceutics, Isfahan Pharmaceutical Sciences Research Center, Isfahan University of Medical Sciences, Isfahan, Iran.
Department of Pharmaceutics and Novel Drug Delivery Systems Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.
Adv Biomed Res. 2016 Oct 26;5:163. doi: 10.4103/2277-9175.190995. eCollection 2016.
A very simple, sensitive, and accurate high-performance liquid chromatography (HPLC) method with ultraviolet detector was developed and applied to determine ciprofloxacin in human plasma following administration of a gastroretentive formulation developed in our laboratory.
HPLC analysis was performed on a C μ-Bondapack column (250 mm × 3.9 mm) using acetonitrile: potassium dihydrogen phosphate solution 0.1 M (20:80, v/v, pH 3) at a flow rate of 1.5 ml/min and eluate was monitored at 276 nm. After addition of phenacetin as internal standard, plasma samples were treated with 0.1 M phosphate buffer (pH: 7) and followed by extraction with dichloromethane. The method was validated for linearity, precision, accuracy, limit of quantitation (LOQ), robustness, stability, and applied in bioavailability studies of our developed gastroretentive formulation in healthy volunteers.
The calibration curves were linear over the concentration range 0.025-4 μg/ml with the detection limit of 15 ng/ml. Accuracy % were within 93-115 and the coefficient of variance % ranged from 0.20 to 12.8. The very low LOQ (25 ng/ml) allowed avoiding fluorometric detection which is more expensive and is not available in all laboratories. Ciprofloxacin was stable in samples with no evidence of degradation during 3 freeze-thaw cycles and 3 months storage at -70°C.
This validated HPLC method was successfully used for the determination of ciprofloxacin in human plasma following oral administration of controlled release formulation, conventional immediate-release tablets and when administered concomitantly with divalent and trivalent cations such as aluminum-, magnesium-, or calcium-containing products under which the bioavailability of ciprofloxacin is significantly reduced.
开发了一种非常简单、灵敏且准确的带紫外检测器的高效液相色谱（HPLC）方法，并将其应用于测定在我们实验室开发的胃滞留制剂给药后人体血浆中的环丙沙星。
HPLC分析在C μ - Bondapack柱（250 mm × 3.9 mm）上进行，使用乙腈：0.1 M磷酸二氢钾溶液（20:80，v/v，pH 3），流速为1.5 ml/min，在276 nm处监测洗脱液。加入非那西丁作为内标后，血浆样品用0.1 M磷酸盐缓冲液（pH：7）处理，然后用二氯甲烷萃取。该方法针对线性、精密度、准确度、定量限（LOQ）、稳健性、稳定性进行了验证，并应用于我们开发的胃滞留制剂在健康志愿者中的生物利用度研究。
校准曲线在0.025 - 4 μg/ml浓度范围内呈线性，检测限为15 ng/ml。准确度百分比在93 - 115之间，变异系数百分比范围为0.20至12.8。极低的定量限（25 ng/ml）使得无需采用更昂贵且并非所有实验室都具备的荧光检测。环丙沙星在样品中稳定，在3次冻融循环和在 - 70°C储存3个月期间无降解迹象。
这种经过验证的HPLC方法成功用于测定口服控释制剂、传统速释片剂后以及与二价和三价阳离子（如含铝、镁或钙的产品）同时给药时人体血浆中的环丙沙星，在这些情况下环丙沙星的生物利用度会显著降低。
