Singh Gurinder, Pai Roopa S, Pandit Vinay
Department of Pharmaceutics, Faculty of Pharmacy, Al-Ameen College of Pharmacy, Bangalore, Karnataka, India.
J Adv Pharm Technol Res. 2012 Apr;3(2):130-5. doi: 10.4103/2231-4040.97296.
A simple, accurate, precise, sensitive, and reproducible high-performance liquid chromatography method was developed for the determination of Resveratrol (trans-3, 4',5-trihydroxystilbene) in human plasma using liquid-liquid extraction. Caffeine was employed as an internal standard (IS). However, little information is known about its distribution in the organism generally because of the lack of accurate and precise detection methods. The chromatographic separation was achieved on a Phenomenex C18 column (250 mm × 4.6 mm, 5 μm) at room temperature in isocratic mode, and the column effluent was monitored by UV detector at 306 nm. The mobile phase used was methanol: phosphate buffer (pH 6.8 adjusted with 0.5% (v/v) orthophosphoric acid solution in Milli-Q water) (63:37%, v/v) at a flow rate of 1.0 ml/min. Nominal retention times of trans-resveratrol and IS were 3.94 and 7.86 minutes, respectively. Limits of detection and Limits of quantification of trans-resveratrol were 0.006 μg/ml and 0.008 μg/ml, respectively. This method was linear over the range of 0.010 to 6.4 μg/ml with regression coefficient greater than 0.9998. The inter- and intra-day precisions in the samples, 0.010, 3.2 and 6.4 μg/ml of trans-resveratrol was in the range 0.63 to 2.12% relative standard deviation (RSD) and 0.46 to 1.02% RSD, respectively. Resveratrol was found to be stable for a period of 15 days on storage at -20°C. The method was found to be precise, accurate, and specific during the study.
建立了一种简单、准确、精密、灵敏且可重现的高效液相色谱法，采用液-液萃取法测定人血浆中的白藜芦醇（反式-3,4',5-三羟基茋）。使用咖啡因作为内标（IS）。然而，由于缺乏准确精密的检测方法，关于其在生物体内的分布情况所知甚少。色谱分离在室温下于Phenomenex C18柱（250 mm×4.6 mm，5μm）上以等度模式进行，柱流出物通过紫外检测器在306 nm处进行监测。所用流动相为甲醇：磷酸盐缓冲液（用Milli-Q水中0.5%（v/v）正磷酸溶液调节pH至6.8）（63:37%，v/v），流速为1.0 ml/min。反式白藜芦醇和内标的标称保留时间分别为3.94分钟和7.86分钟。反式白藜芦醇的检测限和定量限分别为0.006μg/ml和0.008μg/ml。该方法在0.010至6.4μg/ml范围内呈线性，回归系数大于0.9998。样品中反式白藜芦醇浓度为0.010、3.2和6.4μg/ml时，日间和日内精密度的相对标准偏差（RSD）分别在0.63%至2.12%和0.46%至1.02%范围内。发现白藜芦醇在-20°C储存15天内稳定。在研究过程中发现该方法精密、准确且具有特异性。
