高效液相色谱法同时测定市售醒酒护肝产品中8种化合物

Determination of 8 components in healthy food for anti-hangover and hepatoprotection by high performance liquid chromatography

目的建立同时分析醒酒护肝产品中儿茶素、表儿茶素、表没食子儿茶素、表儿茶素没食子酸酯、表没食子儿茶素没食子酸酯、二氢杨梅素、甘草酸和甘草次酸等8种化合物的高效液相色谱法,并与实验室已建立的毛细管电泳法进行方法学比较。方法样品经甲醇-水(4∶1,V/V)超声提取30 min,10 000 r/min离心10 min,取上清液过滤膜后液相色谱进样,采用C_(18)柱(5μm×250 mm×4.6 mm)进行梯度洗脱,流速为0.8 mL/min,色谱柱温度为30℃,采用可变波长检测器在210 nm处对儿茶素类及二氢杨梅素进行检测,250 nm处对甘草酸及甘草次酸进行检测。结果在优化的条件下,8种组分在各自的线性范围内相关系数(r)≥0.9996,检出限和定量限分别为0.07~1.25μg/g(S/N=3)和0.22~4.18μg/g(S/N=10),日内精密度和日间精密度分别为0.26%~1.95%和1.17%~3.89%,加标回收率为86.15%~98.61%。结论该高效液相色谱法灵敏度高、重复性好,可用于醒酒护肝产品的质量控制。

Objective To develop a simple and sensitive high performance liquid chromatographic method for simultaneous determination of catechin hydrate,epicatechin,epigallocatechin, epicatechin gallate, epigallocatechin gallate, dihydromyricetin,glycyrrhizic acid and glycyrrhetinic acid in healthy food for anti-hangover and hepatoprotection,and compare with the capillary electrophoresis method established by our laboratory. Methods The samples were ultrasonically extracted by using methanol-water（ 4∶ 1,V/V） for 30 minutes and then centrifuged at 10 000 r/min for 10 minutes. The supernatant was filtered and injected into the HPLC system and then separated on a C（18） column（ 5 μm × 250 mm × 4. 6 mm） at 30℃ with gradient elution at a flow rate of 0. 8mL/min. Catechins and dihydromyricetin were detected at the wavelength of 210 nm,glycyrrhizic acid and glycyrrhetinic acid were detected at 250 nm. Results Under the optimal analytical conditions,the peak area of each analyte and its concentration had agood correlation within the linear range（ r ≥ 0. 9996）. The limits of detection and quantification of the method were 0. 07-1. 25 μg/g（ S/N = 3） and 0. 22-4. 18 μg/g（ S/N = 10）,respectively. The intra-and inter-day relative standard deviations（ RSDs）of the mixed standard solution were 0. 26%-1. 95% and 1. 17%-3. 89%,respectively.The spiked recoveries of the analytes were 86. 15%-98. 61%. Conclusion The established method is sensitive and reliable,and could be used for quality control of the healthy food for anti-hangover and hepatoprotection.