UPLC法同时测定云南重楼栽培品中11种皂苷的含量

Simultaneous determination of eleven steroidal saponins in cultivated Paris polyphylla Smith var. yunnanensis(Franch.)Hand.-Mazz. by UPLC

目的:建立超高效液相色谱法同时测定云南重楼栽培品中伪原薯蓣皂苷、伪原纤细薯蓣皂苷、重楼皂苷Ⅶ、重楼皂苷D、重楼皂苷H、重楼皂苷Ⅵ、重楼皂苷Ⅱ、薯蓣皂苷、纤细薯蓣皂苷、重楼皂苷Ⅰ、重楼皂苷Ⅴ共11种甾体皂苷的含量。方法:采用CORTECS UPLC C_(18)(2.1 mm×100 mm,1.6μm)色谱柱,以乙腈(A)-水(B)为流动相,梯度洗脱(0~18 min,30%A→50%A;18~28 min,50%A),流速0.3 mL·min^(-1),检测波长203 nm,柱温30℃,进样量1μL。结果:伪原薯蓣皂苷、伪原纤细薯蓣皂苷、重楼皂苷Ⅶ、重楼皂苷D、重楼皂苷H、重楼皂苷Ⅵ、重楼皂苷Ⅱ、薯蓣皂苷、纤细薯蓣皂苷、重楼皂苷Ⅰ、重楼皂苷Ⅴ线性范围分别为2.006~401.2、2.024~404.8、1.972~394.4、1.912~382.4、1.896~379.2、1.772~354.4、2.056~411.2、2.016~403.2、1.970~394.0、1.736~347.2、1.756~351.2μg·mL^(-1);平均回收率(n=6)分别为98.73%、100.30%、101.90%、99.52%、98.33%、100.25%、98.57%、97.70%、101.63%、100.16%和98.80%,RSD分别为2.09%、2.41%、1.77%、2.74%、1.50%、2.35%、2.06%、1.76%、2.82%、2.05%和1.86%。云南重楼栽培品中上述11种甾体皂苷的含量分别为0~0.874、0~0.764、1.066~5.144、0~0.327、1.192~1.952、0~0.455、2.442~6.182、0.394~0.954、0~0.681、2.182~10.105、0~0.674 mg·g^(-1)。结论:该方法可用于同时测定云南重楼栽培品中11种甾体皂苷的含量,为云南重楼药材的质量评价提供参考。

Objective： To establish a UPLC method for the simultaneous determination of 11 steroidal saponins, including pseduoprodioscin, pseudoprotogracillin, paris saponin Ⅶ, paris saponin D, paris saponin H, paris saponin VI, paris saponin Ⅱ, dioscin, gracillin, paris saponin Ⅰ and paris saponin V in Rhizomes of cultivated Paris polyphylla Smith var. yunnanensis （ Franch. ） Hand. -Mazz. by UPLC. Methods： UPLC analysis was performed on a Waters CORTECS UPLC C18 column（ 2. 1 mm × 100 mm, 1.6 μm ）,with a mobile phase consisting of acetonitrile （ A ） -water （ B ） at the flow rate of 0. 3 mL· min-1 （ 0-18 min, 30%A →50%A ; 18-28 min, 50%A ）. The detection wavelength was 203 nm and the column temperature was controlled at 30 ℃, and the injection volume was 1 μL. Results： The linear ranges of pseduoprodioscin, pseudoprotogracillin, paris saponin VII, paris saponin D, paris saponin H, paris saponin Ⅵ, paris saponin Ⅱ, dioscin, gracillin, paris saponin I and paris saponin V were 2. 006-401.2 μg· mL-1, 2. 024-404. 8 μg· mL-1, 1. 972-394. 4 μg·mL-1, 1. 912-382. 4 μg· mL-1, 1. 896-379. 2 μg· mL-1, 1. 772-354.4 μg· mL-1, 2. 056-411.2 μg· mL-1, 2. 016-403.2 μg· mL-1, 1. 970- 394.0 μg· mL-1, 1. 736-347.2 μg·mL-1 and 1. 756-351.2 μg· mL-1, respectively. The average recoveries of the 11 steroidal saponins were 98.73%（ RSD=2.09% ）, 100. 30%（ RSD=2. 41% ）, 101.90%（ RSD=1. 77% ）, 99.52% （ RSD=2.74% ）, 98. 33% （ RSD=1. 50% ）, 100. 25% （ RSD=2. 35% ）, 98.57% （ RSD=2. 06% ）, 97.70% （ RSD=I. 76% ）, 101.63% （ RSD=2. 82% ）, 100. 16% （ RSD=2.05% ） and 98.80% （ RSD=1. 86% ）, respectively. The content ranges of the above 11 steroidal saponins in Rhizomes of cultivated Paris polyphylla Smith vat. yunnanensis （ Franch. ） Hand. -Mazz. were 0-0. 874 mg· g-l, 0-0. 764 mg· g-1, 1. 066-5. 144 mg· g-1, 0-0. 327 mg· g-1 1. 192-1. 952 mg· g-1 ,0-0. 455 mg·g-1, 2. 442-6. 182 mg·g-1 0. 394-0. 954 mg· g-1, 0-0. 681 mg · g-l, 2. 182-10. 105 mg · g-1and 0-0. 674 mg · g-1, respectively. Conclusion： The established method could determine 11 steroidal saponins in cultivated Paris polyphylla Smith var. yurmanensis （Franch.） Hand. - Mazz, thus provided reference for qualtiy evaluation of the medicinal material.