水解工艺对苦荞提取物中槲皮素含量的影响及其药代动力学研究

Effect of hydrolysis process on quercetin content in Tartary buckwheat extract and its pharmacokinetics

目的:以苦荞提取物为研究对象,通过正交实验设计优化苦荞提取物中芦丁被酸水解为槲皮素的工艺条件,并结合药代动力学研究探讨酸水解工艺对苦荞提取物中芦丁和槲皮素吸收的影响。方法:以盐酸百分含量、水解时间、乙醇百分含量、水解温度作为因素设计四因素三水平正交实验优化苦荞提取物中芦丁酸水解为槲皮素的工艺条件,以该工艺进行苦荞提取物酸水解,并在水解第1、2、4、6 h取样得到不同槲皮素含量的水解产物1、2、3、4。分别灌胃大鼠200 mg/kg的苦荞提取物及各水解产物,并在不同时间点采血,以黄芩素为内标,高效液相色谱(HPLC)测定血浆中槲皮素的浓度,DAS 2.0计算主要药代动力学参数。结果:苦荞提取物芦丁酸水解的最佳工艺参数为盐酸百分含量0.91%、水解时间6 h、乙醇百分含量50%、水解温度70℃,在该工艺条件下64.11%芦丁转化为槲皮素。苦荞提取物、水解产物1、2、3、4槲皮素的AUC_(0-t)分别为(17.604±5.422)、(42.175±9.435)、(87.917±19.347)、(116.706±46.256)、(178.509±49.478)mg/L·h,C_(max)分别为(2.743±1.217)、(7.109±2.603)、(12.438±3.197)、(15.727±4.68)、(20.044±5.736)mg/L。与苦荞提取物相比,其水解产物1、2、3、4的药代动力学参数AUC_(0-t)、C_(max)具有显著性差异(p<0.05)。结论:苦荞提取物经酸水解后部分芦丁转化为槲皮素,芦丁和槲皮素在大鼠体内的累计吸收量也相应提高。

Objective：To investigate the effect of hydrolysis process on quercetin content in Tartary Buckwheat extract and explore the acid hydrolysis influence on absorption of rutin and quercetin.Methods：Hydrochloric acid percentage,hydrolysis time,ethanol percentage and hydrolysis temperature were used to design a four-factor and three-level orthogonal test to optimize the acid hydrolysis process of rutin to quercertin,and hydrolyzate samples 1,2,3,4 with different quercetin contents were prepared at 1,2,4,6 h by this process.After oral administration of tartary buckwheat extract and hydrolysis samples（200 mg/kg）,the blood samples were collected at the given time points.The concentration of quercetin was determined by HPLC,and the main pharmacokinetic parameters were calculated by DAS 2.0 software package.Results：The optimum parameters of rutin acid hydrolysis：The percentage of hydrochloric acid content,hydrolysis time,ethanol concentration and hydrolysis temperature were 0.91%,6 h,50%and 70℃,respectively.And 64.11%of rutin was hydrolyzed into quercetin under this process conditions.The AUC（0-t）of Tartary buckwheat extract,hydrolyzate samples 1,2,3and 4 equaled（17.604±5.422）,（42.175±9.435）,（87.917±19.347）,（116.706±46.256）and（178.509±49.478）mg/L·h,respectively,while the C（max）of each sample was（2.743±1.217）,（7.109±2.603）,（12.438±3.197）,（15.727±4.68）and（20.044±5.736）mg/L,respectively.Compared to the Tartary buckwheat extract,the AUC（0-t）and C（max）of hydrolyzate samples 1,2,3 and 4 was significantly increased（p0.05）.Conclusions：With part of rutin converted to quercetin after acid hydrolysis,their cumulative absorption in rats was improved correspondingly.