摘要
目的建立测定牡丹花粉破壁率及其2种单萜苷含量的HPLC法,同时比较不同类型花粉在相同提取条件下2种单萜苷的含量差异。方法采用细胞计数法,以完整花粉个数为评价指标,测定牡丹花粉的破壁率。色谱条件:Globalsil C18色谱柱(250 mm×4.6 mm,5.0μm);流动相:乙腈(A)-1 mL·L^-1甲酸溶液(B)。梯度洗脱:0~3 min,84%B^81%B;3~8 min,81%B;8~11 min,81%B^79%B;11~21 min,79%B;21~40 min,70%B^60%B。流速:1.0 mL·min^-1;柱温:35℃;检测波长:240 nm;进样量:10μL。结果牡丹花粉粒可在显微镜下观察清晰,花粉粒个数与花粉的质量呈正相关,在0.0021~0.0061 g·mL^-1范围内线性关系良好,可用于牡丹花粉显微计数。牡丹花粉经破壁后的破壁率为89.16%。芍药苷和氧化芍药苷分别在质量浓度为6.3~202.8(r1=0.9999)和6.4~205.8μg·mL^-1(r2=0.9998)范围内与各自峰面积呈良好的线性关系,平均回收率分别为99.78%和99.56%,RSD值分别为1.23%和1.27%;精密度、稳定性和重复性实验的RSD值均小于3.0%。经同一方法处理后,2种类型花粉中氧化芍药苷含量相差不大,而芍药苷含量有显著差别。结论利用细胞计数法测定牡丹花粉破壁率方法简单、稳定;其液相方法准确、可靠,重复性好,可用于牡丹花粉中芍药苷和氧化芍药苷的定量分析;破壁对花粉中有效成分的溶出起到促进作用。
Objective To establish a method to determine wall-broken rate and an HPLC method for content determination of paeoniflorin and oxypaeoniflorin in peony pollen,and to compare the content difference of 2 kinds of monoterpene glycosides in different types of pollen under same extraction conditions.Methods Cytometry was used to investigate the influence of ultra-pulverization on peony pollen,and wall-broken rate of peony pollen was also determined.The HPLC analysis was carried out on a Globalsil C18 column(250 mm×4.6 mm,5.0μm)with mobile phase consisted of acetonitrile(A)-1 mL·L^-1formic acid(B)with gradient elution(0-3 min,84%B-81%B;3-8 min,81%B;8-11 min,81%B-79%B;11-21 min,79%B;21-40 min,70%B-60%B).The flow rate was 1.0 mL·min^-1;column temperature was 35℃;detection wavelength was 240 nm;injection volume was 10μL.Results The pollen grains of peony pollen can be clearly observed under a microscope.The regression analysis indicated the weight(0.0021-0.0061 g·mL^-1)of peony pollen and the number of complete cells correlated significantly.The wall-broken rate of peony pollen was 89.16%.The paeoniflorin and oxypaeoniflorin showed good linearity in the ranges of 6.3-202.8(r1=0.9999)and 6.4-205.8μg·mL^-1(r2=0.9998),respectively.The average recoveries were 99.78%(RSD=1.23%)and 99.56%(RSD=1.27%),respectively.RSDs of precision,stability and reproducibility tests were all less than 3.0%.After the same treatment,the content of oxypaeoniflorin in 2 types of pollen was not significantly different,but the content of paeoniflorin was significantly different.Conclusion Cell counting method on the peony pollen is convenient and stable,and the HPLC method is simple and reproducible,and can be used for the content determination of paeoniflorin and oxypaeoniflorin in peony pollen.The broken wall promoted the dissolution of effective components in pollen.
作者
武俊生
李捷
王静静
李雨欣
罗欢欢
林奋
王四旺
WU Junsheng;LI Jie;WANG Jingjing;LI Yuxin;LUO Huanhuan;LIN Fen;WANG Siwang(Department of Life Science and Medicine,Northwest University,Xi′an 710069,China;Department of Chinese Materia Medica and Natural Medicines,Air Force Medical University,Xi′an 710032,China;Shaanxi Fengdan Zhengyuan Biological Technology Limited Company,Xi′an 710077,China)
出处
《西北药学杂志》
CAS
2020年第4期494-498,共5页
Northwest Pharmaceutical Journal
基金
陕西省生物医药重点实验室后补助项目(编号:2018SZS41)。
关键词
牡丹花粉
破壁率
含量测定
HPLC法
peony pollen
wall-broken rate
content determination
HPLC
