基于多元统计分析的川产道地药材江油附子和川乌中6种生物碱含量比较研究
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摘要
目的建立UPLC法同时测定附子和川乌中苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、新乌头碱、乌头碱和次乌头碱6种生物碱含量的方法,并采用多元统计分析方法比较附子和川乌中的6种生物碱的含量差异。方法采用0.05mol/m L盐酸水超声提取附子和川乌中的生物碱类成分,BEH C18(100 mm×2.1 mm,1.7μm)色谱柱;乙腈-水梯度洗脱;体积流量0.1 mL/min,柱温35℃;进样体积4μL;检测波长235 nm。采用差异显著性分析、效/毒值分析、毒性成分指数分析、相关性分析、聚类热图分析、主成分分析和TOPSIS分析进行数据处理,综合评价附子和川乌的质量。结果方法学考察结果符合规定,附子和川乌中苯甲酰次乌头原碱、新乌头碱、次乌头碱和MDAs含量差异极显著,苯甲酰新乌头原碱和苯甲酰乌头原碱含量差异显著,乌头碱和双酯型生物碱(DDAs)含量差异不显著。以6种生物碱为评价指标,15号附子样品质量最优,7号川乌样品质量最优。结论建立的方法操作简单、准确可靠,具有良好的精密度、稳定性、重复性,可用于附子和川乌中生物碱类化合物的定量测定;效/毒值、毒性成分指数的运用能更精确、简便地实现附子和川乌的质量控制;基于多元统计分析可更准确地进行药材质量评价。可为进一步完善《中国药典》附子和川乌中酯型生物碱的分析测定提供参考,为附子和川乌的质量控制及评价提供科学依据。
Objective To establish a UPLC method for the simultaneous determination of benzoylmesaconitine, benzoylaconitine,benzoylhypaconitine, mesaconitine, hypaconitine and aconitinein Aconiti Lateralis Radix Praeparata(ALRP) and Aconiti Radix(AR),and to compare the content difference of six alkaloids in the samples with multivariate statistical analysis. Methods Alkaloids in ALRP and AR were extracted with 0.05 mol/mL hydrochloric acid solution by ultrasonic extraction. The UPLC method was performed on a BEH C18(100 mm × 2.1 mm, 1.7 μm) through a gradient elution of acetonitrile and water at a flow rate of 0.1 mL/min with column temperature at 35 ℃. The injection volume was set at 4.0 μL, and the detection wavelength was set at 235 nm. Difference significance analysis, activity ratio analysis, toxic constituents index analysis, correlation analysis, hierarchical cluster heatmap analysis, principal component analysis, and TOPSIS analysis were used for data processing to comprehensively evaluate the quality of ALRP and AR. Results The method was in accordance with the regulations. Compared with the content of six alkaloids in ALRP and AR, significant difference existed in mesaconitine, benzoylhypaconitine, aconitine, and monoester diterpenoid alkaloids(MDAs); Significant difference existed in benzoylmesaconitine and benzoylaconitine; There were no significant difference in hypaconitine and diester-diterpenoid alkaloids(DDAs).Taking the six alkaloids as indexes, the sample No. 15 of ALRP was the best in quality and the samples No. 7 of AR was the best in quality.Conclusion The established method is simple, accurate and reliable with good precision, repeatability and stability, which can be used for the simultaneous determination of six alkaloids in ALRP and AR. It is much more accurate and convenient to conduct the quality evaluation by using activity ratio, toxic constituents index and multivariate statistical analysis. The study could provide a reference for improving analysis and determination of ester-type alkaloids in ALRP and AR in ChP, and providing a reference for quality control and evaluation of ALRP and AR.
Objective To establish a UPLC method for the simultaneous determination of benzoylmesaconitine, benzoylaconitine,benzoylhypaconitine, mesaconitine, hypaconitine and aconitinein Aconiti Lateralis Radix Praeparata(ALRP) and Aconiti Radix(AR),and to compare the content difference of six alkaloids in the samples with multivariate statistical analysis. Methods Alkaloids in ALRP and AR were extracted with 0.05 mol/mL hydrochloric acid solution by ultrasonic extraction. The UPLC method was performed on a BEH C18(100 mm × 2.1 mm, 1.7 μm) through a gradient elution of acetonitrile and water at a flow rate of 0.1 mL/min with column temperature at 35 ℃. The injection volume was set at 4.0 μL, and the detection wavelength was set at 235 nm. Difference significance analysis, activity ratio analysis, toxic constituents index analysis, correlation analysis, hierarchical cluster heatmap analysis, principal component analysis, and TOPSIS analysis were used for data processing to comprehensively evaluate the quality of ALRP and AR. Results The method was in accordance with the regulations. Compared with the content of six alkaloids in ALRP and AR, significant difference existed in mesaconitine, benzoylhypaconitine, aconitine, and monoester diterpenoid alkaloids(MDAs); Significant difference existed in benzoylmesaconitine and benzoylaconitine; There were no significant difference in hypaconitine and diester-diterpenoid alkaloids(DDAs).Taking the six alkaloids as indexes, the sample No. 15 of ALRP was the best in quality and the samples No. 7 of AR was the best in quality.Conclusion The established method is simple, accurate and reliable with good precision, repeatability and stability, which can be used for the simultaneous determination of six alkaloids in ALRP and AR. It is much more accurate and convenient to conduct the quality evaluation by using activity ratio, toxic constituents index and multivariate statistical analysis. The study could provide a reference for improving analysis and determination of ester-type alkaloids in ALRP and AR in ChP, and providing a reference for quality control and evaluation of ALRP and AR.
引文
[1]中国药典[S].一部.2015.
[2]滕爱武,孙巧英,刘蕾.川乌与附子不可混用[J].时珍国医国药,2000,11(10):900.
[3]钱长敏,宋兆辉,张兰兰,等.四川道地产区乌头药材不同部位6种生物碱含量对比研究[J].中国中药杂志,2013,38(17):2761-2767.
[4]徐婷,钟凌云,罗诣涵.不同姜制附子中6种生物碱含量的比较[J].中成药,2017,39(12):2555-2559.
[5]刘科兰,刘荣华,黄慧莲.附子不同炮制品三种单酯型生物碱的含量差异研究[J].时珍国医国药,2016,27(4):864-866.
[6]舒晓燕,余马,侯大斌,等.不同栽培区附子生物碱含量的比较研究[J].安徽农业科学,2014,42(30):10519-10520.
[7]黄志芳,易进海,陈东安,等.制川乌HPLC特征图谱研究和6种酯型生物碱的含量测定[J].药物分析杂志,2011,31(2):217-221.
[8]赵纳,侯大斌,刘向鸿.不同炮制方法对附子中乌头总碱和双酯型生物碱含量的影响[J].中药材,2011,34(1):39-42.
[9]Xie Y,Jiang Z H,Zhou H,et al.Simultaneous determination of six Aconitum alkaloids in proprietary Chinese medicines by high-performance liquid chromatography[J].J Chromatogr A,2005,1093(1):195-203.
[10]黄志芳,易进海,唐小龙,等.乌头类药材酯型生物碱含量测定方法的优化[J].中国实验方剂学杂志,2013,19(14):101-104.
[11]黄志芳,杨昌林,张意涵,等.UPLC-MS/MS法测定市售附子中的13种生物碱[J].华西药学杂志,2016,31(5):505-508.
[12]唐小龙,易进海,夏燕莉,等.不同炮制方法对附子6种酯型生物碱含量的影响[J].中国实验方剂学杂志,2013,19(21):96-100.
[13]张定堃,王伽伯,杨明,等.中药品质整合评控实践:附子品质综合指数[J].中国中药杂志,2015,40(13):2582-2588.
[14]张定堃.中药质量整合评控模式和方法研究[D].成都:成都中医药大学,2016.
[15]谭鹏,张海珠,张青,等.UPLC法同时测定大黄中10个蒽醌衍生物的含量[J].中草药,2018,49(4):928-934.
[16]张悦,朱荫,叶火香,等.不同产地香茶的主要化学成分含量的差异分析[J].食品科学,2017,38(22):184-191.
[17]谢俊大,孙磊.重楼药材多指标含量分析及化学计量学综合质量评价[J].药物分析杂志,2015,35(9):1585-1590.
[18]张文霖.主成分分析在SPSS中的操作应用[J].市场研究,2005(12):31-34.
[19]侯小龙,熊苗苗,朱红莲,等.不同品系子莲荷叶中荷叶碱和槲皮素的含量测定及品质评价[J].中国医院药学杂志,2013,33(8):666-668.
[20]杨正明,刘哲,邓秋林,等.四川道地产区江油附子和川乌中4种核苷类成分含量测定及多元统计分析[J].中草药,2018,49(23):5657-5664.
[2]滕爱武,孙巧英,刘蕾.川乌与附子不可混用[J].时珍国医国药,2000,11(10):900.
[3]钱长敏,宋兆辉,张兰兰,等.四川道地产区乌头药材不同部位6种生物碱含量对比研究[J].中国中药杂志,2013,38(17):2761-2767.
[4]徐婷,钟凌云,罗诣涵.不同姜制附子中6种生物碱含量的比较[J].中成药,2017,39(12):2555-2559.
[5]刘科兰,刘荣华,黄慧莲.附子不同炮制品三种单酯型生物碱的含量差异研究[J].时珍国医国药,2016,27(4):864-866.
[6]舒晓燕,余马,侯大斌,等.不同栽培区附子生物碱含量的比较研究[J].安徽农业科学,2014,42(30):10519-10520.
[7]黄志芳,易进海,陈东安,等.制川乌HPLC特征图谱研究和6种酯型生物碱的含量测定[J].药物分析杂志,2011,31(2):217-221.
[8]赵纳,侯大斌,刘向鸿.不同炮制方法对附子中乌头总碱和双酯型生物碱含量的影响[J].中药材,2011,34(1):39-42.
[9]Xie Y,Jiang Z H,Zhou H,et al.Simultaneous determination of six Aconitum alkaloids in proprietary Chinese medicines by high-performance liquid chromatography[J].J Chromatogr A,2005,1093(1):195-203.
[10]黄志芳,易进海,唐小龙,等.乌头类药材酯型生物碱含量测定方法的优化[J].中国实验方剂学杂志,2013,19(14):101-104.
[11]黄志芳,杨昌林,张意涵,等.UPLC-MS/MS法测定市售附子中的13种生物碱[J].华西药学杂志,2016,31(5):505-508.
[12]唐小龙,易进海,夏燕莉,等.不同炮制方法对附子6种酯型生物碱含量的影响[J].中国实验方剂学杂志,2013,19(21):96-100.
[13]张定堃,王伽伯,杨明,等.中药品质整合评控实践:附子品质综合指数[J].中国中药杂志,2015,40(13):2582-2588.
[14]张定堃.中药质量整合评控模式和方法研究[D].成都:成都中医药大学,2016.
[15]谭鹏,张海珠,张青,等.UPLC法同时测定大黄中10个蒽醌衍生物的含量[J].中草药,2018,49(4):928-934.
[16]张悦,朱荫,叶火香,等.不同产地香茶的主要化学成分含量的差异分析[J].食品科学,2017,38(22):184-191.
[17]谢俊大,孙磊.重楼药材多指标含量分析及化学计量学综合质量评价[J].药物分析杂志,2015,35(9):1585-1590.
[18]张文霖.主成分分析在SPSS中的操作应用[J].市场研究,2005(12):31-34.
[19]侯小龙,熊苗苗,朱红莲,等.不同品系子莲荷叶中荷叶碱和槲皮素的含量测定及品质评价[J].中国医院药学杂志,2013,33(8):666-668.
[20]杨正明,刘哲,邓秋林,等.四川道地产区江油附子和川乌中4种核苷类成分含量测定及多元统计分析[J].中草药,2018,49(23):5657-5664.