金樱子配方颗粒提取工艺优化及质量标准、指纹图谱研究

Optimization of the Extraction Technology of Rosa laevigata Formula Granules and Study on Its Quality Standard and Fingerprint

目的:优化金樱子配方颗粒的提取工艺,并初步建立该配方颗粒的质量标准及指纹图谱。方法:以金樱子药材出膏率和金樱子多糖提取率为指标,以提取次数、加水量和提取时间为因素,采用正交试验设计优化金樱子配方颗粒的提取工艺并进行验证试验。采用薄层色谱法(TLC)对金樱子配方颗粒进行定性鉴别;采用紫外分光光度法(UV)对金樱子配方颗粒中的金樱子多糖(以D-无水葡萄糖计)进行含量测定。采用高效液相色谱法(HPLC)建立金樱子配方颗粒的指纹图谱;以8号峰为参照,绘制10批金樱子配方颗粒样品的HPLC图谱,采用《中药色谱指纹图谱相似度评价系统》(2012版)进行相似度评价,确定共有峰。结果:优选提取工艺为8倍量水,回流提取2次,每次2 h;金樱子药材平均出膏率为51.07%(RSD=0.58%,n=3)、金樱子多糖平均提取率为22.17%(RSD=0.58%,n=3)。金樱子配方颗粒样品TLC图斑点清晰,分离度好。UV法测定D-无水葡萄糖检测质量浓度线性范围为0.010 8~0.054 2 mg/m L(r=0.999 1);金樱子多糖定量限为0.070 3 mg,检测限为0.014 0 mg;精密度、稳定性、重复性试验的RSD均小于2%;加样回收率为97.3%~100.8%(RSD=1.33%,n=9)。10批金樱子配方颗粒样品的HPLC图谱有8个共有峰,相似度均大于0.937;经验证,该10批金樱子配方颗粒样品的HPLC图谱与对照指纹图谱具有较好的一致性。结论:金樱子配方颗粒的优化提取工艺稳定、可行;所建的指纹图谱可为金樱子配方颗粒的质量评价提供依据。

OBJECTIVE:To optimize the extraction technology and establish the quality standard and fingerprint of Rosa laevigata formula granules primarily.METHODS:Using the yield of extract and extract rate of R.laevigata polysaccharide as index,the extraction technology of R.laevigata formula granules was optimized by orthogonal test with water amount,extraction times and extracting time as factors.Validation test was also conducted.TLC was used to identity R.laevigata formula granule qualitatively.The content of R.laevigata polysaccharide(by D-glucosum anhydricum)in R.laevigata formula granules was determined quantitatively by UV spectrophotometry.HPLC method was adopted to establish the fingerprint of R.laevigata formula granules.Using No.8 peak as reference,HPLC chromatograms of 10 batches of R.laevigata formula granules were drawn.The similarity of samples was evaluated by using TCM Chromatographic Fingerprint Similarity Evaluation System(2012 edition)to determine common peak.RESULTS:The optimal extraction technology was as follows as 8-fold water,extracting for 2 times,2 h each time.The average yield was 51.07%(RSD=0.58%,n=3)and the average extraction rate of polysaccharide in R.laevigata was 22.17%(RSD=0.58%,n=3).TLC spots of R.laevigata formula granules were clear and well-separated.In UV test,the linear range of D-glucosum anhydricum were 0.010 8-0.054 2 mg/mL(r=0.999 1),respectively;for R.laevigata polysaccharide,the limit of quantitation was 0.070 3 mg,and the limit of detection was 0.014 0 mg;RSDs of precision,stability and reproducibility tests were lower than 2%;average recoveries were 97.3%-100.8%(RSD=1.33%,n=9).There were 8 common peaks in HPLC fingerprints of 10 batches of samples,and the similarity was higher than 0.937.After validation,HPLC chromatograms of 10 batches of sample had good consistency with reference fingerprints.CONCLUSIONS:The optimized extraction technology of R.laevigata formula granules is stable.Established fingerprint can provide reference for quality evaluation of R.laevigata formula granules.