HPLC波长转换法同时测定白芥子饮片中2种活性成分含量

Contents of two active components in Sinapis alba decoction piece determined simultaneously by HPLC wavelength conversion

目的 建立HPLC波长转换法同时测定白芥子饮片中白芥子苷和芥子碱硫氰酸盐含量的方法。方法 对建立的高效液相色谱法进行方法学考察,并对收集的15批样品进行白芥子苷和芥子碱硫氰酸盐含量检测。色谱条件采用Kromasil C_(18)柱(5μm,250 mm×4.6 mm),水(含0.08 mol/L磷酸二氢钾,A)-乙腈(B)作为流动相,梯度洗脱,流速为1 ml/min;0~10 min检测波长为227 nm,10 min以后为326 nm;柱温30℃;进样量5μl。结果 白芥子苷质量浓度在7.485~74.85μg/ml(R^(2)=0.999 5,n=6),芥子碱硫氰酸盐质量浓度在0.864~8.64μg/ml范围内与峰面积线性关系良好(R^(2)=0.999 2,n=6)。精密度、稳定性、重复性试验的RSD<2%;白芥子苷的平均加样回收率为107.99%,RSD为1.00%(n=6),芥子碱硫氰酸盐的平均加样回收率为106.42%,RSD为1.14%(n=6)。结论 本研究建立的高效液相色谱法操作简便、结果准确可靠,可用于同时测定白芥子饮片中白芥子苷和芥子碱硫氰酸盐的含量。

Objective To establish an HPLC wavelength conversion method for the content determination of sinalbin and sinapine thiocyanate in Sinapis alba decoction piece.Methods The HPLC method was established and investigated methodologically.Then the contents of sinalbin and sinapine thiocyanate were determined in 15 batches of collected samples.The chromatographic column was carried out on Kromasil C 18 column(5μm,4.6 mm×250 mm)using water(containing 0.08 mol/L KH 2PO 4,A)-acetonitrile(B)as the flow phase for gradient elution,with the volume flow of 1 ml/min,the detection wavelength was set at 227 nm at 0-10 min and then 326 nm after 10 min,the column temperature was 30℃and the sample volume was 5μl.Results There was a good linear relationship in the range of 7.485-74.85μg/ml for sinalbin(R^(2)=0.9995,n=6),and 0.864-8.64μg/ml for sinapine thiocyanate(R^(2)=0.9992,n=6).The RSDs of precision,stability and repetition tests were all less than 2%.The average recovery of sinalbin was 107.99%and RSD was 1.00%(n=6),and the average recovery of sinapine thiocyanate was 106.42%and RSD was 1.14%(n=6).Conclusion The established HPLC is simple and reliable,and can be used to simultaneously determine the contents of sinalbin and sinapine thiocyanate in Sinapis alba decoction piece.