HPLC-RID法测定腺梗豨莶药材中豨莶精醇等4种化合物的含量

目的建立测定腺梗豨莶中豨莶精醇、16β,17-二羟基-贝壳杉烷-19-羧酸、16α氢-贝壳杉烷-17,19-二羧酸和3',4'-二甲氧基槲皮素含量的高效液相色谱示差折光检测方法(HPLC-RID)。方法以十八烷基硅烷键合相为固定相,流动相为体积分数0.3%甲酸溶液-乙腈(体积比42∶55),流速为1.0 mL·min-1,示差检测器光学元件温度为40℃,柱温30℃。结果豨莶精醇的线性范围为0.202².02 g·L-1,16β,17-二羟基-贝壳杉烷-19-羧酸的线性范围为0.5102.02 g·L-1,16β,17-二羟基-贝壳杉烷-19-羧酸的线性范围为0.510⁵.10 g·L-1,16α氢-贝壳杉烷-17,19-二羧酸的线性范围为0.2985.10 g·L-1,16α氢-贝壳杉烷-17,19-二羧酸的线性范围为0.298².98 g·L-1,3',4'-二甲氧基槲皮素的线性范围为0.1002.98 g·L-1,3',4'-二甲氧基槲皮素的线性范围为0.100¹.00 g·L-1,4种化合物线性关系良好(r>0.999);平均加样回收率在99.2%1.00 g·L-1,4种化合物线性关系良好(r>0.999);平均加样回收率在99.2%¹04.9%内;RSD小于3.4%。结论该方法简便、准确、重复性良好,适用于腺梗豨莶中4种成分的同时测定。

Simultaneous determination of four major constituents in Siegesbeckia pubescens Makino by HPLC-RID

Objective To establish an HPLC-RID method for simultaneous monitoring of four major constituents in Siegesbeckia pubescens Makino. Methods The separation of four compounds w as achieved w ith a Waters Symmetry ShieldTMRP18 column( 250 mm × 4. 6 mm,5 μm) and the mobile phase w as 0. 3% formic acid-acetonitrile( V∶ V = 42∶ 55). The temperatures of the optical element and column w ere 40 ℃ and 30 ℃,respectively. The flow rate w as 1. 0 mL·min- 1. Results This method w as linear for all the four compounds w ith correlation coefficients > 0. 999 0. The calibration curve w as constructed using four calibration samples in the range of 0. 202-2. 02 g·L- 1for darutigenol,0. 510-5. 10 g·L- 1for 16β,17-dihydroxy-kauran-19-oic acid,0. 298-2. 98 g·L- 1for 16α H-kauran-17,19-dioic acid and 0. 100-1. 00 g·L- 1for 3',4'-dimethoxy quercetin. Accuracy w as validated on the basis of quotient of the averaged measurements and the nominal value and it ranged from 99. 2% to 104. 9%. The RSD w ere w ithin 3. 4%. Conclusions The method w hich affords simplicity,high accuracy and excellent precision is successfully applied to determine the four major constituents in Siegesbeckia pubescens Makino simultaneously.