RP-HPLC法测定热感赛比斯坦颗粒中3种生物碱成分的含量

目的：建立热感赛比斯坦颗粒中吗啡、可待因和罂粟碱3种生物碱成分的含量测定方法。方法：采用高效液相法进行定量分析。吗啡含量测定选用C₁₈（250 mm×4.6 mm，5 μm）色谱柱，以乙腈-0.01 mol·L^-1磷酸二氢钾和0.05 mol·L^-1庚烷磺酸钠混合水溶液（pH 3.1）（2∶98）为流动相，流速1 mL·min^-1，检测波长220 nm；柱温28℃。可待因和罂粟碱含量测定选用C₁₈（250 mm×4.6 mm，5 μm）色谱柱，以乙腈（A）-0.01 mol·L^-1磷酸二氢钾和0.05 mol·L^-1庚烷磺酸钠混合水溶液（pH 3.8）（B）为流动相进行梯度洗脱（0~18 min，2% A→40% A；18~25 min，40% A→52% A），流速1 mL·min^-1，检测波长分别为220 nm和257 nm，柱温28℃。结果：吗啡、磷酸可待因和盐酸罂粟碱进样量分别在0.119~23.760、0.057~3.983和0.022~3.906 μg范围内，与峰面积线性关系良好（r=0.999 2~0.999 9），平均回收率（n=6）分别为100.2%、100.0%和100.2%。3批样品中吗啡、可待因和罂粟碱的含量分别为0.030~0.031、0.440~0.444和0.011~0.012 mg·g^-1。结论：该方法可用于热感赛比斯坦颗粒中3种生物碱的含量测定。

Objective: To establish the method for determination of morphine,codeine and papaverine in Regan Saibisitan granules. Methods: High performance liquid chromatography method was employed for the analyses of three alkaloids in Regan Saibisitan granules.Morphine analysis was carried out on C₁₈ column(250 mm×4.6 mm,5 μm)with mobile phase consisting of acetonitrile-0.01 mol·L^-1 monopotassium phosphate and 0.05 mol·L^-1 sodium heptanesulfonate solution(pH 3.1)(2:98).The flow rate was 1.0 mL·min^-1 and the detection wavelength was 220 nm.The column temperature was maintained at 28℃.Analyses of codeine and papaverine were carried out on C₁₈ column(250 mm×4.6 mm,5 μm)with mobile phase consisting of acetonitrile(A) -0.01 mol·L^-1 monopotassium phosphate and 0.05 mol·L^-1 sodium heptanesulfonate solution(pH 3.8)(B)by gradient elution(0-18 min,2%A→40%A;18-25 min,40%A→52%A).The flow rate was 1.0 mL·min -1 and the detection wavelengths were 220 nm and 257 nm.The column temperature was maintained at 28℃. Results: The linear ranges of morphine,codeine phosphate and papaverine hydrochloride were 0.119-23.760 μg(r=0.999 8),0.057-3.983 μg(r=0.999 9)and 0.022-3.906 μg(r=0.999 2),respectively.The average recoveries(n=6)were 100.2%,100.0% and 100.2%,respectively.The contents of morphine,codeine and papaverine in 3 samples were 0.030-0.031 mg·g^-1,0.440-0.444 mg·g^-1 and 0.011-0.012 mg·g^-1,respectively. Conclusion: The established determination method is proved by methodology validation that it can be used for the detection of 3 alkaloids of opium poppy shell and seed in Regan Saibisitan granules.

[1] 中华人民共国卫生部药品标准.维吾尔药分册[S]. 1998:177 Drug Specifications Promulgated by the Ministry of Public Health. PR China. Uigher Medicine[S]. 1998:177
[2] 范蕾,胡剑影,王伟影.汤料中罂粟壳成分的检测[J].海峡药学, 2012,24(10):65 FAN L, HU JY, WANG WY, et al. Determination of alkaloid of poppy shell in soup materials in chafing dish[J]. Strait Pharm J, 2012,24(10):65
[3] 李雕,李进瞳,曾燕,等.不同等级罂粟壳有效成分含量的研究[J].中国中药杂志, 2010, 35(17):2246 LI D, LI JT, ZENG Y, et al. Study on the content of effective components in different grades of poppy[J]. China J Chin Mater Med, 2010, 35(17):2246
[4] 贺金华,薛桂蓬,康雨彤,等.热感赛比斯坦颗粒定性定量方法研究[J].药物分析杂志,2015, 35(1):139 HE JH, XUE GP, KANG YT, et al. Study on qualitative and quantitative methods for the determination of Regansaibisitan granules[J]. Chin J Pharm Anal, 2015, 35(1):139
[5] 中国药典2010年版.二部[S]. 2010:1153 ChP 2010. Vol Ⅱ[S]. 2010:1153
[6] 刘永利,袁浩,李冬梅,等. UPLC法测定罂粟壳中3种生物碱类成分含量[J].中药材, 2011,34(9):1386 LIU YL, YUAN H,LI DM,et al. Determination of 3 alkaloids in the shell of opium poppy by UPLC[J]. J Chin Med Mater, 2011, 34(9):1386
[7] 中国药典2015年版.一部[S]. 2015:369 ChP 2015. Vol Ⅰ[S]. 2015:369
[8] 刘永利,李冬梅,冯丽,等. "一测多评" 法测定咳喘宁片中5种生物碱类成分的含量[J].中药新药与临床药理,2012,23(4):465 LIU YL, LI DM,FENG L, et al. Simultaneous assay of five kinds of alkaloids in Kechuanning tablets by quantitative analysis of multicomponents by single marker[J]. Tradit Chin Drug Res Clin Pharmcol, 2012,23(4):465
[9] 刘文炜,高玉琼,刘建华,等. RP-HPLC法同时测定哮喘片中吗啡、盐酸麻黄碱、磷酸可待因的含量[J].药物分析杂志,2009, 29(5):731 LIU WW, GAO YQ, LIU JH, et al. RP-HPLC simultaneous determination of morphine, ephedrine hydrochloride and codeine phosphate in Xiaochuan tablets[J]. Chin J Pharm Anal, 2009, 29(5):731
[10] 张庆雷,李雪君,刘峰,等.反向离子对高效液相色谱法测定祖卡木颗粒中吗啡的含量[J].广东药学院学报,2008,24(2):112 ZHANG QL,LI XJ, LIU F, et al. Determination of morphine content in Zukamu granules by ion-pair RP-HPLC[J]. J Guangdong Med Coll, 2008, 24(2):112
[11] 孙晓霞,郭凤霞,李锦萍,等. HPLC同时测定复方磷酸可待因糖浆中的有关物质和防腐剂[J].华西药学杂志,2015,30(5):588 SUN XX, GUO FX, LI JP, et al. Simultaneous determination of the related substances and preservatives in compound codeine phosphate syrup by HPLC[J]. West China J Pharm Sci, 2015, 30(5):588
[12] 邱琳,张媛媛,刘莹. RP-HPLC法同时测定人参保肺丸中吗啡、磷酸可待因的含量[J].世界科学技术-中医药现代化, 2014, 16(3):591 QIU L,ZHANG YY,LIU Y,et al. Simultaneous determination of the content of morphine and codeine phosphate by RP-HPLC[J]. World Sci Technol Mod Tradit Chin Med Mater Med,2014,16(3):591
[13] 刘永利,李冬梅,袁浩,等.固相萃取-超高效液相色谱法同时测定肉蔻四神丸中3种有效成分含量[J].中国药学杂志, 2011, 46(22):1754 LIU YL, LI DM, YUAN H, et al. Simultaneous determination of three alkaloids in Roukousishen pills by SPE-UPLC[J]. Chin Pharm J, 2011, 46(22):1754
[14] 李娜,赵斌,黄文华,等.高效液相色谱法测定克咳胶囊中吗啡的含量[J].时珍国医国药, 2005,16(4):321 LI N, ZHAO B, HUANG WH, et al. Determination of morphine in Keke cough capsules by HPLC[J]. Lisizhehn Med Mater Med Res, 2005, 16(4):321
[15] 刘永利,李冬梅,冯丽,等.固相萃取-多波长超高效液相色谱法同时测定咳喘宁中5种生物碱类成分的含量[J].药学学报, 2011, 46(5):581 LIU YL,LI DM,FENG L,et al. Simultaneous determination of 5 kinds of alkaloids in Kechuanning tablets by SPE-UPLC under different UV-vis wavelength[J]. Acta Pharm Sin,2011,46(5):581