Shehzadi Naureen, Hussain Khalid, Khan Muhammad Tanveer, Salman Muhammad, Islam Muhammad
Punjab University College of Pharmacy, University of the Punjab, Allama Iqbal Campus, Lahore, Pakistan.
Punjab University College of Pharmacy, University of the Punjab, Allama Iqbal Campus, Lahore, Pakistan / School of Pharmaceutical Sciences, University Sains Malaysia, Pulau Penang, Malaysia.
Pak J Pharm Sci. 2017 Sep;30(5):1767-1777.
The absence of chromophore and/or conjugated system, prerequisite for UV and florescent light detection, or absorbance at very low wavelength necessitates the development of simple and reliable methods for the determination of amikacin sulphate. Therefore, the present study describes for the first time dynamics of the drug derivatization using ninhydrin reagent and development and validation of a simple RP-HPLC method, using diode array detector (DAD). The variables such as heating time, heating type, drug-reagent ratio, reagent composition and storage temperature of the derivative were optimized. The analyte and aqueous ninhydrin solution upon heating for 2.00-5.00 min produced the colored drug-derivative which was stable for one month at refrigeration. The derivatized drug (20.00μL) was eluted through a column - Eclipse DB-C (5.00 µm, 4.60×150.00 mm), maintained at 25°C- using isocratic mobile phase comprising water and acetonitrile (70:30, v/v) at a flow rate of 1.00 mL/min, and detected at 400 nm. The method was found to be reliable (98.08-100.72% recovery), repeatable (98.02-100.72% intraday accuracy) and reproducible (98.47-101.27% inter day accuracy) with relative standard deviation less than 5%. The results of the present study indicate that the method is easy to perform, specific and sensitive, and suitable to be used for the determination of amikacin sulphate in bulk and pharmaceutical preparations using less expensive/laborious derivatization.
由于缺乏发色团和/或共轭体系（紫外线和荧光检测的先决条件），或在极低波长下有吸收，因此需要开发简单可靠的方法来测定硫酸阿米卡星。因此，本研究首次描述了使用茚三酮试剂进行药物衍生化的动力学过程，以及使用二极管阵列检测器（DAD）开发和验证一种简单的反相高效液相色谱法。对诸如加热时间、加热方式、药物与试剂比例、试剂组成以及衍生物的储存温度等变量进行了优化。分析物与茚三酮水溶液加热2.00 - 5.00分钟后生成有色药物衍生物，该衍生物在冷藏条件下可稳定保存一个月。衍生化药物（20.00μL）通过Eclipse DB - C柱（5.00 µm，4.60×150.00 mm）洗脱，柱温保持在25°C，使用由水和乙腈（70:30，v/v）组成的等度流动相，流速为1.00 mL/min，并在400 nm波长处进行检测。该方法被证明是可靠的（回收率为98.08 - 100.72%）、可重复的（日内准确度为98.02 - 100.72%）和可重现的（日间准确度为98.47 - 101.27%），相对标准偏差小于5%。本研究结果表明，该方法操作简便、特异性强、灵敏度高，适用于使用成本较低/操作简便的衍生化方法来测定原料药和药物制剂中的硫酸阿米卡星。
