HPLC法同时测定大黄炮制品中10种化学成分的含量

Simultaneous Determination of 10 Kinds of Chemical Components in Processed Products of Rhei Radix et Rhizoma

目的：建立同时测定生大黄、酒大黄、熟大黄、大黄炭、醋大黄中没食子酸、儿茶素、番泻苷B、芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚、大黄酚-1-D-葡萄糖苷、大黄素-8-D．葡萄糖苷含量的方法，并分析其差异。方法：采用高相液相色谱法。色谱柱为Hypersil C18,流动相为甲醇-0．2％乙酸（梯度洗脱），流速为1．0ml／min，检测波长为260nm，柱温为25℃，进样量为10ul。结果：没食子酸、儿茶素、番泻苷B、芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚、大黄酚．1-D-葡萄糖苷、大黄素-8-D．葡萄糖苷检测进样量线性范围分别为0．2525～4．0400gg（r=0．9996）、0．6000～9．6000gg（r=0．9996）、0．2974～4．7584（r=0．9999）、0．0018～0．0288gg（r=0．9999）、0．005O～0．0800ug（4=0．9999）、0．019O～0．3040Ixg（，=0．9998）、0．3802～6．0832ug（r：0．9997）、0．0082～0．1312ktg（r=0．9998）、0．1260～2．0160μg（r=0．9996）、0．1113～1．7808jig（r=0．9998）；精密度、稳定性、重复性试验的RSD〈3．O％；加样回收率分别为96．17％～97．21％（RSD=1．67％，n=6）、97．60％～100．54％（RSD=2．55％，n=6）、99．45％～101．32％（RSD=1．63％，n=6）、95．31％～98．19％（RSD=2．42％，n=6）、98．99％～100．35％（RSD=1．86％，n=6）、98．95％～101．21％（RSD=2．17％，n=6）、99．81％～100．62％（RSD=1．66％，n=6）、96．78％～98．52％（RSD=1．99％，n=6）、97．80％～100．14％（RSD=3．32％，n=6）、97．40％～101．24％（RSD=2．89％，n=6）。与生大黄比较，酒大黄、醋大黄、大黄炭中的没食予酸、儿茶素、番泻苷B和蒽醌类成分含量减少，熟大黄中儿茶素、番泻苷B和蒽醌类成分含量减少，在大黄炭中未检测到儿茶素、番泻苷B、大黄素-1-D-葡萄糖苷、芦荟大黄素、大黄酸。结论：该方法操作简便，精密度、稳定性、重复性良好，可用于大黄炮制品中lO种化学成分的同时测定；大黄不同炮制品中10种化学成分含量均有明显差异。

OBJECTIVE： To establish a method for the contents of gallic acid, catechin, sennosides B, aloe-emodin, rhein, emodin, chrysophanol, physcion, chrysophanol -1-O- glucoside and emodin -8-O- glucoside in Rhei Radix et Rhizoma, Jiu Rhei Radix et Rhizoma, Shu Rhei Radix et Rhizoma, Rhei Radix et Rhizoma tan, Cu Rhei Radix et Rhizoma, and analyze the differ- ences. METHODS： HPLC was performed on the column was Hypersil C18 with mobile phase of methanol - 0.2% acetic acid（gradi- ent elution） at a flow rate of 1.0 ml/min, the detection wavelength was 260 nm, column temperature was 25 %, injection volume was 10μl. RESULTS： The linear range was 0.252 5-4.040 0 pg for gallic acid （r=0.999 6）, 0.600 0-9.600 0 p.g for catechin （r= 0.999 6）,0.297 4-4.758 4 gg for sennosides B（r=0.999 9）,0.001 8-0.028 8 p.g for aloe-emodin（r=0.999 9）,0.005 0-0.080 0 μg for rhein （r=0.999 9）,0.019 0-0.304 0 gg for emodin （r=0.999 8）,0.380 2-6.083 2 gg for chrysophanol（r=0.999 7）,0.008 2-0.131 2 μg for physcion （r=0.999 8）, 0.126 0-2.016 0 μg for chrysophanol-1-O-glucoside（r=0.999 6） and 0.111 3-1.780 8 I.tg for emo- din -8-O-glucoside （r=0.999 8） ; RSDs of precision, stability and reproducibility tests were lower than 3.0% ; recoveries were 96.17% -97.21% （RSD=l.67%, n=6）, 97.60% -100.54% （RSD=2.55%, n=6）, 99.45% -101.32% （RSD=l.63%, n=6）, 95.31% -98.19% （RSD=2.42% , n=6） , 98.99% -100.35% （RSD=l.86% , n=6） , 98.95% -101.21% （RSD=2.17% , n=6） , 99.81%-100.62% （RSD=l.66% ,n=6）, 96.78%-98.52% （RSD=l.99% ,n=6）, 97.80%-100.14% （RSD=3.32% ,n=6） and 97.40 %-101.24 % （RSD = 2.89 %, n = 6 ）. Compared with Sheng Rhei Radix et Rhizoma, the contents of gallic acid, catechin, sen- nosides B and anthraquinones in Cu Rhei Radix et Rhizoma, Jiu Rhei Radix et Rhizoma and Rhei Radix et Rhizoma tan decreased. The contents of catechin, sennosides B and anthraquinones in Shu Rhei Radix et Rhizoma. Catechin, sennosides B, chrysopha- nol -1-O- glucoside, aloe-emodin and rhein were not detected in Dahuang tan. CONCLUSIONS： The method is simple with good precision, stability and reroducibility, and can be used for the simultaneous determination of 10 chemical components in processed products of Rhei Radix et Rhizoma; there were significant differences in contents of 10 chemical components in processed prod- ucts of Rhei Radix et Rhizoma.