Determination of trans-resveratrol in mouse liver by high performance liquid chromatography

高效液相色谱法测定小鼠肝脏中反式-白藜芦醇含量(英文)

To develop a sensitive high performance liquid chromatography （HPLC） assay for the determination of trans-resveratrol in mouse liver. The whole liver of a mouse was removed from the body, homogenated, and extracted by ethyl acetate. The organic layer was isolated and evaporated to dryness, the residue was reconstituted in 0.2 mL mobile phase for centrifugation, and 50 uL of the supernatant was injected into the/-IPLC instrument. The sample was separated on a Shimadzu ODS column （150 mm × 4.6 mm, 5 um） at 35 ℃ and detected by ultraviolet （UV） detector at the wavelength of 305 nm. The mobile phase consisted of methanol and 0.1 mol/L acetic acid （4：6, v/v） with the flow-rote at 1 mL/min. The limit of detection was 3.0 ng/g in liver homogenate with a signal/noise ratio of 3：1. The linear range of the calibration curve was 5.0-120.0 ng/g. The mean recoveries at the concentrations of 6, 10 and 80 ng/g were 102%, 96.0% and 91.5%, respectively. The RSDs for inter- and intra-day assays were less than 5%. Compared with other reported methods, this method was faster and more sensitive. It was also proved to be of good linearity, selectivity, accuracy and precision, and can be efficiently applied to the pharmacoldnetic study of trans-resveratrol in mouse liver.

建立—灵敏的HPLC法,用于测定小鼠肝脏中反式-白藜芦醇含量。取小鼠全部肝脏制成匀浆,用乙酸乙酯提取。分离有机层并挥干,残渣用0.2mL流动相溶解,离心,取上清液50μL注入高效液相色谱仪。样品在岛津ODS柱(150 mm×4.6mm,5μm)上分离,柱温35℃,检测波长305nm,流动相为甲醇-0.1 mol/L乙酸溶液(4:6,v/v),流速为1 mL/min。以信噪比3:1计,白藜芦醇在小鼠肝脏中的检测限为3.0 ng/g;标准曲线的线性范围为5.0～120.0ng/g;浓度为6、10及80ng/g的平均回收率分别为102%、96.0%及91.2%;日内、日间精密度的RSD均小于5%。与已报道的其它白藜芦醇生物样品测定方法相比较,本法更为快速与灵敏,并具有良好的线性、选择性、准确度与精密度,可用于小鼠肝脏中白藜芦醇的药物动力学研究。