Amanolahi Farjad, Mohammadi Ali, Kazemi Oskuee Reza, Nassirli Hooriyeh, Malaekeh-Nikouei Bizhan
Department of Drug and Food Control, School of Pharmacy, Tehran University of Medical Science, Tehran, Iran.
Department of Medical Biotechnology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Avicenna J Phytomed. 2017 Sep-Oct;7(5):444-453.
This study was designed to develop and validate a new reversed-phase high-performance liquid chromatography (RP-HPLC) method based on Q (R) International Conference on Harmonization (ICH) guideline for determination of curcumin in pharmaceutical samples.
The HPLC instrument method was optimized with isocratic elution with acetonitrile: ammonium acetate (45:55, v/v, pH 3.5), C18 column (150 mm×4.6 mm×5 µm particle size) and a flow rate of 1 ml/min in ambient condition and total retention time of 17 min. The volume of injection was set at 20 µl and detection was recorded at 425 nm. The robustness of the method was examined by changing the mobile phase composition, mobile phase pH, and flow rate.
The method was validated with respect to precision, accuracy and linearity in a concentration range of 2-100 µg/ml. The limit of detection (LOD) and limit of quantification (LOQ) were 0.25 and 0.5 µg/ml, respectively. The percentage of recovery was 98.9 to 100.5 with relative standard deviation (RSD) < 0.638%.
The method was found to be simple, sensitive and rapid for determination of curcumin in pharmaceutical samples and had enough sensitivity to detect degradation product of curcumin produced under photolysis and hydrolysis stress condition.
本研究旨在根据国际协调会议（ICH）指南开发并验证一种新的反相高效液相色谱（RP-HPLC）方法，用于测定药物样品中的姜黄素。
采用乙腈：醋酸铵（45:55，v/v，pH 3.5）等度洗脱，C18柱（150 mm×4.6 mm×5 µm粒径），在环境条件下流速为1 ml/min，总保留时间为17 min，对HPLC仪器方法进行优化。进样体积设定为20 µl，检测波长为425 nm。通过改变流动相组成、流动相pH值和流速来考察该方法的稳健性。
该方法在2-100 µg/ml浓度范围内进行了精密度、准确度和线性验证。检测限（LOD）和定量限（LOQ）分别为0.25和0.5 µg/ml。回收率为98.9%至100.5%，相对标准偏差（RSD）<0.638%。
该方法用于测定药物样品中的姜黄素简单、灵敏、快速，且具有足够的灵敏度来检测姜黄素在光解和水解应激条件下产生的降解产物。
