不同炮制品配伍的青娥丸中12种指标成分含量变化研究
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摘要
目的建立HPLC同时测定青娥丸中12种指标成分的方法,研究不同炮制品配伍的青娥丸中指标性成分含量变化。方法色谱柱:Purospher~ STAR LPRP-C_(18)endcapped(250 mm×4.6 mm,5μm);流动相:0.1%甲酸水-乙腈梯度洗脱;流速:1.0 m L·min~(-1);检测波长:240 nm;进样量:10μL;柱温:30℃。对不同炮制品配伍的青娥丸中指标性成分进行含量测定。结果在上述色谱条件下,青娥丸中12个成分分离度良好,加样回收率范围96.04%~104.07%,RSD均小于2.95%。盐炙品青娥丸中补骨脂素、异补骨脂素、补骨脂定、补骨脂二氢黄酮甲醚、Corlifol A、补骨脂酚、异补骨脂查尔酮7种成分含量明显下降(P<0.05,P<0.01,P<0.001),其他成分无明显变化。结论本研究所建立的方法简单,准确性和稳定性较好,可用于青娥丸的质量控制。盐炙品配伍的青娥丸中补骨脂素、异补骨脂素等成分与生品配伍的青娥丸比较含量明显下降。
Objective To establish a HPLC method for the simultaneous determination of 12 components in Qing'e pills, and to study the contents of indicative components in Qing 'e pills composed of herbs processed by different methods. Methods The separation was achieved on Purospher~STAR LPRP- C_(18) endcapped column(250 mm×4.6mm, 5 μm). The mobile phase composed of acetonitrile and 0.1 % formic acid aqueous solution, and a gradient elution program at flow rate of 1.0 m L·min~(-1)was applied. The UV detection wavelength was set at 240 nm and the injection volume was 10 μL. The column temperature was maintained at 30 ℃. Results Under the above chromatographic conditions,good separation was achieved among the 12 components. The sample recovery rate was within the range of96.04 % to 104.07 % with RSD values less than 2.95 %. The results showed that the contents of psoralen,isopsoralen, psoralidin, bavachinin, corylifol A and bakuchiol in the salt- processed herb compatibility weresignificantly reduced(P < 0.05 compared with the raw herb compatibility),but the contents of the others components had no remarkable changes. Conclusion The established HPLC method is accurate,stable and available,which can be suitable for the quality control of Qing'e pills. The pills composed of salt- processed herbs have lower contents of indicative components such as psoralen and isopsoralen than the pills composed of raw herbs.
Objective To establish a HPLC method for the simultaneous determination of 12 components in Qing'e pills, and to study the contents of indicative components in Qing 'e pills composed of herbs processed by different methods. Methods The separation was achieved on Purospher~STAR LPRP- C_(18) endcapped column(250 mm×4.6mm, 5 μm). The mobile phase composed of acetonitrile and 0.1 % formic acid aqueous solution, and a gradient elution program at flow rate of 1.0 m L·min~(-1)was applied. The UV detection wavelength was set at 240 nm and the injection volume was 10 μL. The column temperature was maintained at 30 ℃. Results Under the above chromatographic conditions,good separation was achieved among the 12 components. The sample recovery rate was within the range of96.04 % to 104.07 % with RSD values less than 2.95 %. The results showed that the contents of psoralen,isopsoralen, psoralidin, bavachinin, corylifol A and bakuchiol in the salt- processed herb compatibility weresignificantly reduced(P < 0.05 compared with the raw herb compatibility),but the contents of the others components had no remarkable changes. Conclusion The established HPLC method is accurate,stable and available,which can be suitable for the quality control of Qing'e pills. The pills composed of salt- processed herbs have lower contents of indicative components such as psoralen and isopsoralen than the pills composed of raw herbs.
引文
[1]国家药典委员会.中华人民共和国药典(一部)[M].北京:中国医药科技出版社,2015:1024-1025.
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[5]张紫佳,陈洁,赵俊铭,等.超高效液相色谱法测定青娥丸中松脂醇二葡萄糖苷的含量[J].色谱,2010,28(8):805-808.
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[7]艾路,马群,邱落,等.高效液相色谱法侧定加味青峨丸中补骨脂素、异补骨脂素的含量[J].世界科学技术(中药现代化),2010,12(5):808-810.
[8]颜翠萍,吴育,翁泽斌,等.盐制对补骨脂中主要化学成分的影响[J].中成药,2013,35(11):2470-2474.
[9]Wen ZB,Yan CP,Wu Y,et al.Determination of Ten Components in Eucommia ulmoides Oliver by High-Performance Liquid Chromatography with Dual Detectors[J].Analytical Letters,2014,47(12):1978-1986.
[10]林芳.杜仲治疗骨质疏松的有效成分筛选及质量控制的研究[D].成都:成都中医药大学,2012.
[11]Don MJ,Lin LC,Chion WF.Neobavaisoflavone stimulates osteogenesis via p38-mediated up-regulation of transcription factors and osteoid genes expression in MC3T3-E1 cells[J].Phytomedicine,2012,19:551-561.
[12]Li WD,Yan CP,Wu Y,et al.Osteoblasts proliferation and differentiation stimulating activities of the main components of Fructus Psoraleae corylifoliae[J].Phytomedicine,2014,21:400-405.
[13]Tang DZ,Yang F,Yang Z,et al.Psoralen stimulates osteoblast differentiation through activation of BMP signaling[J].Biochem Biophys Res Commun,2011,405:256-261.
[14]Wong RWK,Bakr RA.Effect of psoralen on bone formation[J].J Orthop Res,2011,29:158-164.
[15]颜翠萍,翁泽斌,吴育,等.青娥丸盐炙品与生品抗去卵巢诱导的骨质疏松效应的比较研究[J].南京中医药大学学报,2014,30(5):438-442.
[16]Weng ZB,Gao QQ,Wang F,et al.Positive skeletal effect of two ingredients of Psoralea corylifolia L.on estrogen deficiency-induced osteoporosis and the possible mechanisms of action[J].Molecular and Cellular Endocrinology,2015(417):103-113.
[17]Wang YF,Liu YN,Xiong W,et al.A UPLC-MS/MS method for in vivo and in vitro pharmacokinetic studies of psoralenoside,isopsoralenoside,psoralen and isopsoralen from Psoralea corylifolia extract[J].Journal of Ethnopharmacology,2014(151):609-617.
[2]颜翠萍,翁泽斌,李伟东,等.青娥丸盐炙品与生品抗去卵巢诱导的骨质疏松效应的比较研究[J].南京中医药大学学报,2014,30(5):438-442.
[3]徐晓娟,沈霖,杨艳萍,等.青娥丸对绝经后骨质疏松症患者骨密度和骨转换标志物的影响[J].中国中医骨伤科杂志,2013,21(6):8-10.
[4]赵光,沈霖,杨艳萍.青娥丸对绝经后骨质疏松症患者骨密度、血清水平MMP-2及骨代谢指标的影响[J].中西医结合研究,2012,4(3):114-117.
[5]张紫佳,陈洁,赵俊铭,等.超高效液相色谱法测定青娥丸中松脂醇二葡萄糖苷的含量[J].色谱,2010,28(8):805-808.
[6]陈洁,徐颖,张紫佳,等.青娥丸质量标准研究-君药杜仲的含量测定[J].中国实验方剂学杂志,2012,18(20):112-114.
[7]艾路,马群,邱落,等.高效液相色谱法侧定加味青峨丸中补骨脂素、异补骨脂素的含量[J].世界科学技术(中药现代化),2010,12(5):808-810.
[8]颜翠萍,吴育,翁泽斌,等.盐制对补骨脂中主要化学成分的影响[J].中成药,2013,35(11):2470-2474.
[9]Wen ZB,Yan CP,Wu Y,et al.Determination of Ten Components in Eucommia ulmoides Oliver by High-Performance Liquid Chromatography with Dual Detectors[J].Analytical Letters,2014,47(12):1978-1986.
[10]林芳.杜仲治疗骨质疏松的有效成分筛选及质量控制的研究[D].成都:成都中医药大学,2012.
[11]Don MJ,Lin LC,Chion WF.Neobavaisoflavone stimulates osteogenesis via p38-mediated up-regulation of transcription factors and osteoid genes expression in MC3T3-E1 cells[J].Phytomedicine,2012,19:551-561.
[12]Li WD,Yan CP,Wu Y,et al.Osteoblasts proliferation and differentiation stimulating activities of the main components of Fructus Psoraleae corylifoliae[J].Phytomedicine,2014,21:400-405.
[13]Tang DZ,Yang F,Yang Z,et al.Psoralen stimulates osteoblast differentiation through activation of BMP signaling[J].Biochem Biophys Res Commun,2011,405:256-261.
[14]Wong RWK,Bakr RA.Effect of psoralen on bone formation[J].J Orthop Res,2011,29:158-164.
[15]颜翠萍,翁泽斌,吴育,等.青娥丸盐炙品与生品抗去卵巢诱导的骨质疏松效应的比较研究[J].南京中医药大学学报,2014,30(5):438-442.
[16]Weng ZB,Gao QQ,Wang F,et al.Positive skeletal effect of two ingredients of Psoralea corylifolia L.on estrogen deficiency-induced osteoporosis and the possible mechanisms of action[J].Molecular and Cellular Endocrinology,2015(417):103-113.
[17]Wang YF,Liu YN,Xiong W,et al.A UPLC-MS/MS method for in vivo and in vitro pharmacokinetic studies of psoralenoside,isopsoralenoside,psoralen and isopsoralen from Psoralea corylifolia extract[J].Journal of Ethnopharmacology,2014(151):609-617.