HPLC测定片仔癀中4种成分的含量

Determination of Four Compounds in Pien Tze Huang by HPLC

目的建立测定片仔癀中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1及牛磺胆酸钠的含量。方法采用Hypersil BDSC18(4.6 mm×250 mm,5μm);流动相:乙腈(A)-0.5%磷酸溶液(B)梯度洗脱:0~40 min(20%A^40%A),40~90 min(40%A^90%A),90~100 min(90%A);柱温:30℃;流速:1.0 mL.min-1;检测波长:203 nm。结果测得三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1及牛磺胆酸钠的回收率分别为100.3%,101.6%,103.3%,100.6%。线性范围分别是:三七皂苷R10.346 4~8.66μg(r=0.9963),人参皂苷Rg10.432 2~10.805μg(r=0.9964),人参皂苷Rb10.4224~10.56μg(r=0.9999),牛磺胆酸钠0.448~11.2μg(r=0.999 6)。结论采用高效液相色谱梯度洗脱能将片仔癀中的皂苷及胆汁酸较好的分离检测,方法准确可靠,重复性好,结果稳定,可作为该产品质量控制的方法。

OBJECTIVE To develop a HPLC gradient elution method for the determination of notoginsenoside R1, ginsenoside RG1, ginsenoside Rb1 and sodium taurocholic acid in Pien Tze Huang. METHODS Pien Tze Huang was separated on a Hypersil BDS C18 column（4. 6 mm × 250 mm,5 um）and detected at 203 nm by gradient elution at the flow rate of 1 mL · min^-1 using acetonitrile -0. 5 % phosphoric acid （ 2 0 ： 8 0 ） as the mobile phase A （ acetonitrile ） , and 0.5 % phosphoric acid as the mobile phase B. The gradient program were as follows：A：0 min,20% ; 0 -40 min ,20% -40% ;40 -90 min,40% -90% ;90 - 100 min ,90%. Column temperature was at 30 ℃. RESULTS The recoveries of notoginsenoside R1 , ginsenoside Rg1 , ginsenoside Rb1 and sodium taurocholie acid were 100. 3%, 101.6%, 103.3% and 100. 6%, respectively. The good linearities were obtained in the ranges of 0. 346 4 - 8.66 ug （r =0. 996 3） for ginsenoside Rb1, 0. 432 2 - 10. 805 ug（r =0. 996 4）, 0. 422 4 - 10. 56 ug（r =0. 999 9）, 0. 448 - 11.2 ug（r =0. 999 6） for sodium taurocholic acid. CONCLUSION The HPLC gradient elution method had been successfully applied to determine the contents of saponin and bile acids in Pien Tze Huang. It can be used for quality control of Pien Tze Huang.