正交试验优选银桑降糖颗粒中桑叶的提取工艺

Optimization of Extraction Process for Mori Folium Leaves in Yinsang Jiangtang Granules by Orthogonal Test

摘要:

目的: 优选银桑降糖颗粒中桑叶的提取工艺,为该制剂的研发提供参考. 方法: 以芦丁提取量为指标,采用L₉(3⁴)正交试验考察乙醇体积分数及用量、提取时间对桑叶醇提取工艺的影响;以1-脱氧野尻霉素(DNJ)相对提取率为指标,通过L₉(3³)正交试验考察加水量及煎煮时间对桑叶水煎煮工艺的影响.采用HPLC测定芦丁含量,检测波长358 nm,流动相甲醇(A)-0.5%磷酸溶液(B)梯度洗脱(0~5 min,30%A;5~10 min,30%~35% A;10~15 min,35%~40%A;15~18 min,40%~50%A);利用芴甲氧羰酰氯柱前衍生HPLC-UV测定DNJ含量. 结果: 最佳醇提工艺为加20倍量60%乙醇提取1次,提取时间1.5 h;芦丁提取量1.22 mg·g^-1.最佳水煎煮工艺为分别加30,25倍量水煎煮2次,煎煮时间依次为1.0,0.5 h;DNJ相对提取率88.06%. 结论: 优选的提取工艺稳定可行,可兼顾桑叶中黄酮类、生物碱类成分的提取.

Abstract:

Objective: To optimize extraction process of Mori Folium leaves in Yinsang Jiangtang granules. Method: Taking extracting amount of rutin as index, L₉(3⁴) orthogonal design was adopted to optimize alcohol extraction technology with extraction time,ethanol amount and concentration as factors.With relative yield of 1-deoxynojirimycin(DNJ) as index, L₉(3³) orthogonal design was used to optimize water extraction technology by taking solid-liquid ratio and extraction time as factor.The content of rutin was determined by HPLC,detection wavelength was 358 nm,mobile phase consisted of methanol(A)-0.5% phosphoric acid solution(B) for gradient elution(0-5 min,30%A;5-10 min,30%-35%A;10-15 min,35%-40%A;15-18 min,40%-50%A).The contents of DNJ were detected by precolumn derivation HPLC-UV method with FMOC-Cl as reagent. Result: Optimum alcohol extraction press was as following:extracted twice with 20 times the amount of 60% ethanol for 1.5 h;extracting amount of rutin was 1.22 mg·g^-1.Optimum water extraction process was extracted twice for 1.0,0.5 h with 30,25 times the amount of water;relative yield of DNJ was 88.06%. Conclusion: These optimized extraction technologies are stable and feasible,it can balance extraction of flavonoids and alkaloids in Mori Folium.