HPLC—QAMS同时测定复方黄芩片中6种活性成分的含量(1)
中圖分类号 R284.1；R927 文献标志码 A 文章编号 1001-0408(2018)11-1510-05
DOI 10.6039/j.issn.1001-0408.2018.11.17
摘 要 目的：建立同时测定复方黄芩片中6种活性成分含量的高效液相色谱(HPLC)-一测多评法(QAMS)。方法：采用HPLC法，以黄芩苷为内参物，分别计算其与虎杖苷、汉黄芩苷、黄芩素、穿心莲内酯、脱水穿心莲内酯的相对校正因子(RCF)，通过RCF计算复方黄芩片中上述5种成分的含量(计算值)，同时采用外标法测定这6种成分的含量(实测值)，比较计算值与实测值的差异。色谱柱为ZORBAX Eclipse XDB C18，流动相为乙腈-0.05%磷酸溶液(梯度洗脱)，流速为1.0 mL/min，检测波长为306 nm(虎杖苷)、225 nm(穿心莲内酯)、275 nm(黄芩苷、汉黄芩苷、黄芩素)、254 nm(脱水穿心莲内酯)，柱温为25 ℃，进样量为10 μL。结果：虎杖苷、黄芩苷、汉黄芩苷、黄芩素、穿心莲内酯、脱水穿心莲内酯的检测质量浓度的线性范围分别为4.555～81.99、47.03～846.5、12.73～229.1、12.88～231.8、2.632～47.38、3.508～63.14 μg/mL(r为0.999 2～0.999 7)；检测限分别为2.25、6.92、1.76、4.69、1.37、1.51 ng，定量限分别为6.89、19.98、5.28、13.81、4.15、4.71 ng；平均加样回收率分别为98.2%、98.8%、97.9%、98.9%、98.8%、98.7%(RSD为0.91%～1.32%，n=9)；精密度、重复性、稳定性(48 h)试验的RSD<2.0%(n=6或n=7)。虎杖苷、汉黄芩苷、黄芩素、穿心莲内酯、脱水穿心莲内酯的RCF分别为0.529、0.506、1.004、0.831、1.087，其计算值和实测值比较差异无统计学意义(P>0.05)。结论：该方法简单、有效、结果准确、节约成本，可用于复方黄芩片中上述6种活性成分含量的同时测定。
关键词 一测多评法；高效液相色谱法；复方黄芩片；相对校正因子；活性成分；含量测定
Content Determination of 6 Active Constituents in Compound Huangqin Tablets by HPLC-QAMS
HU Dan(Suzhou Food and Drug Inspection and Testing Center， Anhui Suzhou 234000， China)
ABSTRACT OBJECTIVE： To establish quantitative analysis of HPLC-quantitative analysis of multi-components by single marker (QAMS) for simultaneous determination of 6 active constituents in Compound huangqin tablets. METHODS： Using baicalin as internal content， HPLC method was used to calculate relative correction factor (RCF) of baicalin to polydatin， wogonoside， baicalein， andrographolide and dehydroandrographolide. The contents of above-mentioned 5 components in Compound huangqin tablets were calculated by using RCF (calculation value). The external standard method (ESM) was used to determine the contents of 6 constituents (measured value). The differences between calculation value and measured value were compared. The determination was performed on ZORBAX Eclipse XDB C18 column with mobile phase consisted of acetonitrile-0.05% phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 306 nm for polydatin， 225 nm for andrographolide， 275 nm for baicalin， wogonoside and baicalein， 254 nm for dehydroandrographolide. The column temperature was set at 25 ℃ and the injection volume was 10 μL. RESULTS： The linear range was 4.555-81.99 μg/mL for polydatin， 47.03-846.5 μg/mL for baicalin， 12.73-229.1 μg/mL for wogonoside， 12.88-231.8 μg/mL for baicalein， 2.632-47.38 μg/mL for andrographolide， 3.508-63.14 μg/mL for dehydroandrographolide (r=0.999 2-0.999 7)， respectively. Limits of detection were 2.25， 6.92， 1.76， 4.69， 1.37， 1.51 ng； limits of quantitation were 6.89， 19.98， 5.28， 13.81， 4.15， 4.71 ng. Average recoveries were 98.2%， 98.8%， 97.9%， 98.9%， 98.8%， 98.7% (RSD=0.91%-1.32%， n=9). RSDs of precision， reproducibility and stability tests (48 h) were lower than 2.0% (n=6 or n=7). RCFs of baicalin to polydatin， wogonoside， baicalein， andrographolide and dehydroandrographolide were 0.529， 0.506， 1.004， 0.831 and 1.087， respectively. There was no statistical significance between calculated values and measured ones (P>0.05). CONCLUSIONS： The method is simple， effective， accurate and cost saving. It can be used for simultaneous determination of 6 active constituents in Compound huangqin tablets., 百拇医药(胡丹)
DOI 10.6039/j.issn.1001-0408.2018.11.17
摘 要 目的：建立同时测定复方黄芩片中6种活性成分含量的高效液相色谱(HPLC)-一测多评法(QAMS)。方法：采用HPLC法，以黄芩苷为内参物，分别计算其与虎杖苷、汉黄芩苷、黄芩素、穿心莲内酯、脱水穿心莲内酯的相对校正因子(RCF)，通过RCF计算复方黄芩片中上述5种成分的含量(计算值)，同时采用外标法测定这6种成分的含量(实测值)，比较计算值与实测值的差异。色谱柱为ZORBAX Eclipse XDB C18，流动相为乙腈-0.05%磷酸溶液(梯度洗脱)，流速为1.0 mL/min，检测波长为306 nm(虎杖苷)、225 nm(穿心莲内酯)、275 nm(黄芩苷、汉黄芩苷、黄芩素)、254 nm(脱水穿心莲内酯)，柱温为25 ℃，进样量为10 μL。结果：虎杖苷、黄芩苷、汉黄芩苷、黄芩素、穿心莲内酯、脱水穿心莲内酯的检测质量浓度的线性范围分别为4.555～81.99、47.03～846.5、12.73～229.1、12.88～231.8、2.632～47.38、3.508～63.14 μg/mL(r为0.999 2～0.999 7)；检测限分别为2.25、6.92、1.76、4.69、1.37、1.51 ng，定量限分别为6.89、19.98、5.28、13.81、4.15、4.71 ng；平均加样回收率分别为98.2%、98.8%、97.9%、98.9%、98.8%、98.7%(RSD为0.91%～1.32%，n=9)；精密度、重复性、稳定性(48 h)试验的RSD<2.0%(n=6或n=7)。虎杖苷、汉黄芩苷、黄芩素、穿心莲内酯、脱水穿心莲内酯的RCF分别为0.529、0.506、1.004、0.831、1.087，其计算值和实测值比较差异无统计学意义(P>0.05)。结论：该方法简单、有效、结果准确、节约成本，可用于复方黄芩片中上述6种活性成分含量的同时测定。
关键词 一测多评法；高效液相色谱法；复方黄芩片；相对校正因子；活性成分；含量测定
Content Determination of 6 Active Constituents in Compound Huangqin Tablets by HPLC-QAMS
HU Dan(Suzhou Food and Drug Inspection and Testing Center， Anhui Suzhou 234000， China)
ABSTRACT OBJECTIVE： To establish quantitative analysis of HPLC-quantitative analysis of multi-components by single marker (QAMS) for simultaneous determination of 6 active constituents in Compound huangqin tablets. METHODS： Using baicalin as internal content， HPLC method was used to calculate relative correction factor (RCF) of baicalin to polydatin， wogonoside， baicalein， andrographolide and dehydroandrographolide. The contents of above-mentioned 5 components in Compound huangqin tablets were calculated by using RCF (calculation value). The external standard method (ESM) was used to determine the contents of 6 constituents (measured value). The differences between calculation value and measured value were compared. The determination was performed on ZORBAX Eclipse XDB C18 column with mobile phase consisted of acetonitrile-0.05% phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 306 nm for polydatin， 225 nm for andrographolide， 275 nm for baicalin， wogonoside and baicalein， 254 nm for dehydroandrographolide. The column temperature was set at 25 ℃ and the injection volume was 10 μL. RESULTS： The linear range was 4.555-81.99 μg/mL for polydatin， 47.03-846.5 μg/mL for baicalin， 12.73-229.1 μg/mL for wogonoside， 12.88-231.8 μg/mL for baicalein， 2.632-47.38 μg/mL for andrographolide， 3.508-63.14 μg/mL for dehydroandrographolide (r=0.999 2-0.999 7)， respectively. Limits of detection were 2.25， 6.92， 1.76， 4.69， 1.37， 1.51 ng； limits of quantitation were 6.89， 19.98， 5.28， 13.81， 4.15， 4.71 ng. Average recoveries were 98.2%， 98.8%， 97.9%， 98.9%， 98.8%， 98.7% (RSD=0.91%-1.32%， n=9). RSDs of precision， reproducibility and stability tests (48 h) were lower than 2.0% (n=6 or n=7). RCFs of baicalin to polydatin， wogonoside， baicalein， andrographolide and dehydroandrographolide were 0.529， 0.506， 1.004， 0.831 and 1.087， respectively. There was no statistical significance between calculated values and measured ones (P>0.05). CONCLUSIONS： The method is simple， effective， accurate and cost saving. It can be used for simultaneous determination of 6 active constituents in Compound huangqin tablets., 百拇医药(胡丹)
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