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目的优化啤酒花中8-异戊烯基柚皮素(8-prenylnaringenin,8-PN)的提取及含量测定方法,并进行验证。方法采用L_8(4~1×2~2)正交试验,料液比为1∶15,利用回流法提取啤酒花中的8-PN,反相高效液相色谱法测定啤酒花中8-PN的含量[色谱分离条件:ODS C18反相柱(4.6 mm×250 mm,0.45μm),用乙腈、1%甲酸进行梯度洗脱,流速:1.2 ml/min,检测波长:288 nm,柱温:30℃]。探讨提取溶剂(甲醇、乙醇、正丁醇、乙酸乙酯)、提取时间(40、60 min)、提取温度(40、60℃)对啤酒花中8-PN提取含量的影响。结果以正丁醇为提取溶剂,料液比为1∶15,利用回流提取,提取温度40℃,时间40 min,测得啤酒花中8-PN的最高提取含量为79.1 mg/kg。该方法的标准曲线相关系数R~2=0.999 8,线性范围为0.001~0.1 mg/ml,检出限为0.122 mg/L;检测浓度为0.03 mg/ml标准液的保留时间和峰面积的相对标准偏差(relative standard deviation,RSD)均小于1%,加标回收率在98%~102%之间。结论优化的方法可简便、准确地测定啤酒花中8-PN含量,为啤酒花中8-PN的进一步分离、纯化提供了实验依据。
Objective To optimize the extraction and determination of 8-prenylnaringenin (8-PN) in hops. Methods The orthogonal design of L_8 (4 ~ 1 × 2 ~ 2) was used. The ratio of solid to liquid was 1:15. The 8-PN in hops was extracted by reflux method. The content of 8-PN in hops was determined by RP-HPLC [Chromatographic separation conditions: ODS C18 reversed-phase column (4.6 mm × 250 mm, 0.45 μm) eluting with a gradient of acetonitrile and 1% formic acid at a flow rate of 1.2 ml / min and a detection wavelength of 288 nm at a column temperature of 30 ° C. ]. The effects of extraction solvent (methanol, ethanol, n-butanol and ethyl acetate), extraction time (40 and 60 min) and extraction temperature (40 and 60 ℃) on the extraction of 8-PN in hops were investigated. Results The n-butanol was used as extraction solvent, the ratio of solid to liquid was 1:15, and the maximum extraction content of 8-PN in hops was 79.1 mg / kg when the extraction temperature was 40 ℃ and the extraction time was 40 min. The calibration curve of this method has a correlation coefficient R 2 = 0.999 8, a linear range of 0.001 ~ 0.1 mg / ml and a detection limit of 0.122 mg / L. The retention time and peak area of the standard solution with a concentration of 0.03 mg / The relative standard deviations (RSDs) were less than 1% and the recoveries were between 98% and 102%. Conclusion The optimized method can be used to determine the content of 8-PN in hops conveniently and accurately, which provides an experimental basis for the further separation and purification of 8-PN in hops.