不同剂量~(60)Co-γ射线辐照对二妙丸中6个有效成分影响的研究
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摘要
目的:建立同时测定二妙丸中苍术素、苍术素醇、盐酸小檗碱、盐酸药根碱、盐酸黄柏碱和盐酸巴马汀6个有效成分含量的测定方法,考察不同剂量60Co-γ射线辐照前后有效成分含量的变化。方法:采用Agilent-C_(18)色谱柱(4.6 mm×250 mm,5μm),以乙腈(A)-0.5%磷酸水溶液(B)(每100 mL加十二烷基硫酸钠0.2 g)为流动相进行梯度洗脱,检测波长分别为280 nm和340 nm,流速1.0 mL·min~(-1),柱温28℃。分别选择辐照剂量为2、5、8 kGy对二妙丸进行辐照,比较辐照前后二妙丸中6个有效成分含量的变化,并进行成组t检验考察其显著情况。结果:苍术素、苍术素醇、盐酸小檗碱、盐酸药根碱、盐酸黄柏碱和盐酸巴马汀分别在0.090 1～2.252 5、0.128 9～3.222 5、0.246 5～6.162 5、0.076 8～1.920 0、0.120 3～3.007 5及0.051 2～1.280 0范围内有良好线性,平均加样回收率分别为99.6%、99.7%、100.5%、98.8%、101.4%及99.1%,RSD分别为1.2%、0.60%、0.50%、0.90%、1.0%和0.80%。经过2、5、8 kGy辐照后,二妙丸所含的有效成分苍术素、苍术素醇、盐酸小檗碱、盐酸药根碱、盐酸黄柏碱和盐酸巴马汀均有变化,经成组t-检验,在超过5 kGy后,苍术素及苍术素醇辐照前后含量变化显著(P<0.05)。结论:所建立的二妙丸活性成分测定方法准确,重复性好,简单实用,可作为二妙丸质控方法。在辐照剂量不超过5 kGy时,各成分变化不显著,为二妙丸辐照灭菌提供了参考。
Objective:To establish an HPLC method for simultaneous determination of 6 active ingredients(atractylodin,atractyoldinol,berberine hydrochloride,jatrorrhizine hydrochloride,phellodendrine hydrochloride and palmatine hydrochloride) in Ermiao pills,and to investigate the effect of ~(60)Co-γ ray irradiation on the effective components. Methods:The Agilent-C_(18) column(4.6 mm×250 mm,5 μm) was adopted and the wavelength of UV detection was 280 nm and 340 nm with the flow rate of 1.0 mL·min~(-1). The mobile phase consisted of acetonitrile(A)-0.5% phosphoric acid(B)(containing 0.2 g of sodium dodecyl sulfate per 100 mL) with a gradient elution. The column temperature was 28 ℃. Doses for irradiation on Ermiao pills were 2 kGy,5 kGy and 8 kGy. The contents of 6 active ingredients before and after irradiation were compared,and a t-test study was used to examine the significance. Results:The linear ranges of atractylodin,atractyoldinol,berberine hydrochloride,jatrorrhizine hydrochloride,phellodendrine hydrochloride and palmatine hydrochloride were 0.090 1-2.252 5 μg,0.128 9-3.222 5 μg,0.246 5-6.162 5 μg,0.076 8-1.920 0 μg,0.120 3-3.007 5μg and 0.051 2-1.280 0 μg,and the average recovery were 99.6%,99.7%,100.5%,98.8%,101.4% and 99.1% with RSDs of 1.2%,0.60%,0.50%,0.90%,1.0% and 0.80%.After irradiation at doses of 2 kGy,5 kGy and 8 kGy,the contents of atractylodin,atractyoldinol,berberine hydrochloride,jatrorrhizine hydrochloride,phellodendrine hydrochloride and palmatine hydrochloride all changed. Results of group t-test indicated significant change(P<0.05) of contents of atractylodin and atractyoldinol after irradiation at doses above 5 kGy. Conclusion:The established method is accurate,repeatable,simple and practical,and can be used for quality control of Ermiao pills. When the radiation dose is below 5 kGy,the changes of each component are not significant,which provides a reference for the sterilization of Ermiao pills.
Objective:To establish an HPLC method for simultaneous determination of 6 active ingredients(atractylodin,atractyoldinol,berberine hydrochloride,jatrorrhizine hydrochloride,phellodendrine hydrochloride and palmatine hydrochloride) in Ermiao pills,and to investigate the effect of ~(60)Co-γ ray irradiation on the effective components. Methods:The Agilent-C_(18) column(4.6 mm×250 mm,5 μm) was adopted and the wavelength of UV detection was 280 nm and 340 nm with the flow rate of 1.0 mL·min~(-1). The mobile phase consisted of acetonitrile(A)-0.5% phosphoric acid(B)(containing 0.2 g of sodium dodecyl sulfate per 100 mL) with a gradient elution. The column temperature was 28 ℃. Doses for irradiation on Ermiao pills were 2 kGy,5 kGy and 8 kGy. The contents of 6 active ingredients before and after irradiation were compared,and a t-test study was used to examine the significance. Results:The linear ranges of atractylodin,atractyoldinol,berberine hydrochloride,jatrorrhizine hydrochloride,phellodendrine hydrochloride and palmatine hydrochloride were 0.090 1-2.252 5 μg,0.128 9-3.222 5 μg,0.246 5-6.162 5 μg,0.076 8-1.920 0 μg,0.120 3-3.007 5μg and 0.051 2-1.280 0 μg,and the average recovery were 99.6%,99.7%,100.5%,98.8%,101.4% and 99.1% with RSDs of 1.2%,0.60%,0.50%,0.90%,1.0% and 0.80%.After irradiation at doses of 2 kGy,5 kGy and 8 kGy,the contents of atractylodin,atractyoldinol,berberine hydrochloride,jatrorrhizine hydrochloride,phellodendrine hydrochloride and palmatine hydrochloride all changed. Results of group t-test indicated significant change(P<0.05) of contents of atractylodin and atractyoldinol after irradiation at doses above 5 kGy. Conclusion:The established method is accurate,repeatable,simple and practical,and can be used for quality control of Ermiao pills. When the radiation dose is below 5 kGy,the changes of each component are not significant,which provides a reference for the sterilization of Ermiao pills.
引文
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[11]潘超,张莉,王玉.HPLC法同时测定二妙丸中黄柏碱、木兰花碱、药根碱、盐酸巴马汀和盐酸小檗碱的含量[J].中国生化药物杂志,2012,33(4):361PAN C,ZHANG L,WANG Y.Simultaneous determination of phellodendrine,magnoflorin,jatrorrhiine,palmatine hydrochloride and berberine hydrochloride in Ermiao pillsby HPLC[J].Chin JBiochem Pharm,2012,33(4):361
[12]郭晓,张伟,贾晓益,等.HPLC双波长切换法测定二妙丸中盐酸小檗碱、苍术素含量[J].贵阳中医学院学报,2017,39(5):31GUO X,ZHANG W,JIA XY,et al.Determination of berberine hydrochloride and atractylodes hydrochloride in Er Miao pills by HPLC dual wavelength switching method[J].J Guiyang Coll Tradit Chin Med,2017,39(5):31
[13]雷蓉,周亚楠,刘亚兰,等.二妙丸HPLC指纹图谱及UPLC-LTQ-Orbitrap成分分析[J].中草药,2017,48(17):3511LEI R,ZHOU YN,LIU YL,et al.HPLC fingerprint and UPLC-LTQ-Orbitrap analysis of Er Miao pills[J].China Tradit Herb Drugs,2017,48(17):3511
[14]黄琦,陈炎明,侴桂新,等.苍术化学指纹图谱建立及其稳定性研究[J].上海中医药杂志,2012,46(10):76HUANG Q,CHEN YM,RU GX,et al.Establishment and stability of chemical fingerprint of Atractylodes rhizome[J].Shanghai JTradit Chin Med,2012,46(10):76
[2]中华人民共和国药典2015年版.一部[S].2015:438ChP 2015.VolⅠ[S].2015:438
[3]吴珊珊,胡麟,龚晓猛,等.HPLC法同时测定不同等级黄柏饮片中多指标成分的含量[J].中国药房,2016,27(15):2135WU SS,HU L,GONG XM,et al.Simultaneous determination of multi index components in different grades of Cortex Phellodendri by HPLC[J].China Pharm,2016,27(15):2135
[4]沈娟,尹莲,段金廒.HPLC法测定黄柏生物碱成分含量及在二妙丸类方中的比较研究[J].中国实验方剂学杂志,2010,16(13):31SHEN J,YIN L,DUAN JA.Determination of alkaloids in Cortex Phellodendri by HPLC and comparative study in two Miao pills[J].Chin J Exp Tradit Med Form,2010,16(13):31
[5]赵森淼,王瑞,侴桂新,等.苍术的定性定量分析方法研究[J].药物分析杂志,2010,30(5):954ZHAO SM,WANG R,RU GX,et al.Study on qualitative and quantitative analysis of Atractylodes[J].Chin J Pharm Anal,2010,30(5):954
[6]陈炎明,陈静,侴桂新.苍术化学成分和药理活性研究进展[J].上海中医药大学学学报,2006,20(4):95CHEN YM,CHEN J,RU GX.Advances in the chemical composition and pharmacological activity of Atractylodes rhizome[J].J Shanghai Univ Tradit Chin Med,2006,20(4):95
[7]雷曦,申鸿.60Co-γ射线辐照贮藏中药材对其主要成分的影响[J].中国药房,2002,13(2):119LEI X,SHEN H.60Co-gamma ray irradiation,the influence of Chinese herbal medicine on its main components[J].China Pharm,2002,13(2):119
[8]孙建宇.60Co-γ射线辐照灭菌在中药及制剂中的应用研究[J].中国药师,2006,9(5):464SUN JY.Application of 60Co-ray irradiation sterilization in Chinese medicine and preparation[J].China Pharm,2006,9(5):464
[9]张世才,李奉勤.辐照灭菌在中药方面的应用研究进展[J].中国药业,2009,18(20):76ZHANG SC,LI FQ.Research progress in application of radiation sterilization in Chinese medicine[J].China Pharm,2009,18(20):76
[10]冯少俊,伍振锋,王雅琪,等.中药灭菌工艺研究现状及问题分析[J].中草药,2015,46(18):2667FENG SJ,WU ZF,WANG YQ,et al.Research status and problem analysis of Chinese medicine sterilization technology[J].China Tradit Herb Drugs,2015,46(18):2667
[11]潘超,张莉,王玉.HPLC法同时测定二妙丸中黄柏碱、木兰花碱、药根碱、盐酸巴马汀和盐酸小檗碱的含量[J].中国生化药物杂志,2012,33(4):361PAN C,ZHANG L,WANG Y.Simultaneous determination of phellodendrine,magnoflorin,jatrorrhiine,palmatine hydrochloride and berberine hydrochloride in Ermiao pillsby HPLC[J].Chin JBiochem Pharm,2012,33(4):361
[12]郭晓,张伟,贾晓益,等.HPLC双波长切换法测定二妙丸中盐酸小檗碱、苍术素含量[J].贵阳中医学院学报,2017,39(5):31GUO X,ZHANG W,JIA XY,et al.Determination of berberine hydrochloride and atractylodes hydrochloride in Er Miao pills by HPLC dual wavelength switching method[J].J Guiyang Coll Tradit Chin Med,2017,39(5):31
[13]雷蓉,周亚楠,刘亚兰,等.二妙丸HPLC指纹图谱及UPLC-LTQ-Orbitrap成分分析[J].中草药,2017,48(17):3511LEI R,ZHOU YN,LIU YL,et al.HPLC fingerprint and UPLC-LTQ-Orbitrap analysis of Er Miao pills[J].China Tradit Herb Drugs,2017,48(17):3511
[14]黄琦,陈炎明,侴桂新,等.苍术化学指纹图谱建立及其稳定性研究[J].上海中医药杂志,2012,46(10):76HUANG Q,CHEN YM,RU GX,et al.Establishment and stability of chemical fingerprint of Atractylodes rhizome[J].Shanghai JTradit Chin Med,2012,46(10):76