HPLC法同时测定不同产地甘西鼠尾草及丹参中8种主要成分的含量
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摘要
目的建立同时测定甘西鼠尾草及丹参中丹参素钠、迷迭香酸、紫草酸、丹酚酸B、二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ和丹参酮ⅡA含量的HPLC法,对比不同产地甘西鼠尾草及丹参中8种成分的含量。方法采用SunFire C_(18)(250 mm×4.6 mm,5μm)色谱柱。流动相为乙腈(A)-0.8 mL·L~(-1)甲酸超纯水溶液(B),梯度洗脱(0～19 min,20%A～60%A;19～49 min,60%A～81%A;49～53 min,81%A～91%A;53～57 min,95%A)。流速:1.0 mL·min~(-1);检测波长:270 nm;柱温:26℃;进样量:2μL。结果质量浓度与峰面积线性关系良好(r>0.971 3),精密度、重复性及回收率均符合要求。在50 min内甘西鼠尾草及丹参中8种成分均可实现完全分离。结论青海不同产地甘西鼠尾草较其他产地甘西鼠尾草在脂溶性成分和水溶性成分上均有较好的优势,与丹参相比主要的差别在于丹酚酸B和迷迭香酸的含量,丹酚酸B的含量较丹参样品低,其余4种脂溶性成分均优于丹参样品。
Objective To establish an HPLC method for the simultaneous determination of 8 compounds including sodium danshensu,rosmarinic acid,lithospermic acid,salvianolic acid B,dihydrotanshinquinone-Ⅰ and cryptotanshinone,tanshinone Ⅰ and tanshinone ⅡA in Salvia przewalskii Maxim and Radix Salviae miltiorrhizae collected from different origins,and to compare the contents.Methods The HPLC separation was performed on an SunFire C_(18)(250 mm×4.6 mm,5 μm) column.The mobile phase A was acetonitrile,and the mobile phase B was acetonitrile-0.8 mL·L~(-1)formic acid.The separation was achieved using the following gradient program:(0-19 min,20%A-60%A;19-49 min,60%A-81%A;49-53 min,81%A-91%A; 53-57 min,95%A),at a flow rate of 1.0 mL·min~(-1).The column temperature was controlled at 26 ℃,the UV detection wavelength was 270 nm,and the injection volume was 2 μL.Results The correlation coefficients were more than 0.971 3.Precision,repeatability and recovery met the requirements.The baseline separation was obtained within 50 min for the 8 compounds.Conclusion Compared with other areas,Salvia przewalskii Maxim produced from Qinghai Province has a better advantage in terms of liposoluble and hydrosoluble constituents.Meanwhile,the main difference between Salvia przewalskii Maxim and Radix Salvia miltiorrhiza is the content of salvianolic acid B and rosmarinic acid.The content of salvianolic acid B in Salvia przewalskii Maxim is lower,but the other 4 contents are higher than Radix Salvia miltiorrhiza.
Objective To establish an HPLC method for the simultaneous determination of 8 compounds including sodium danshensu,rosmarinic acid,lithospermic acid,salvianolic acid B,dihydrotanshinquinone-Ⅰ and cryptotanshinone,tanshinone Ⅰ and tanshinone ⅡA in Salvia przewalskii Maxim and Radix Salviae miltiorrhizae collected from different origins,and to compare the contents.Methods The HPLC separation was performed on an SunFire C_(18)(250 mm×4.6 mm,5 μm) column.The mobile phase A was acetonitrile,and the mobile phase B was acetonitrile-0.8 mL·L~(-1)formic acid.The separation was achieved using the following gradient program:(0-19 min,20%A-60%A;19-49 min,60%A-81%A;49-53 min,81%A-91%A; 53-57 min,95%A),at a flow rate of 1.0 mL·min~(-1).The column temperature was controlled at 26 ℃,the UV detection wavelength was 270 nm,and the injection volume was 2 μL.Results The correlation coefficients were more than 0.971 3.Precision,repeatability and recovery met the requirements.The baseline separation was obtained within 50 min for the 8 compounds.Conclusion Compared with other areas,Salvia przewalskii Maxim produced from Qinghai Province has a better advantage in terms of liposoluble and hydrosoluble constituents.Meanwhile,the main difference between Salvia przewalskii Maxim and Radix Salvia miltiorrhiza is the content of salvianolic acid B and rosmarinic acid.The content of salvianolic acid B in Salvia przewalskii Maxim is lower,but the other 4 contents are higher than Radix Salvia miltiorrhiza.
引文
[1] 曾敏,王雨萌,贾璞,等.HPLC法测定红曲灵芝丹参胶囊中4种成分的含量[J].西北药学杂志,2017,32(6):679-682.
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[6] 林超,刘兆国,钱星,等.丹酚酸B在心血管疾病中药理作用研究进展[J].中国药理学通报,2015,31(4):449-452.
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[9] 程体娟,罗慧英,王巧明,等.甘西鼠尾草注射液与丹参注射液对大鼠皮层神经细胞缺血性损伤保护作用的比较研究[J].中国药理与临床,2005,21(3):34-36.
[10] 程体娟,杨志勇.甘西鼠尾草注射液和丹参注射液抗大鼠急性脑缺血和抗脂质过氧化作用的比较[J].中国临床药理学与治疗学,2003,8(1):23-26.
[11] 叶剑.丹参的药用成分与药理作用探析[J].陕西中医学院学报,2012,35(5):71-73.
[12] 杨阳,朱斌,吴志军,等.甘西鼠尾草提取物对正常大鼠全血粘度影响及对水负荷大鼠利尿作用[J].中国医院药学杂志,2012,32(10):751-754.
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[14] 杨阳,王林,张毕奎.丹参中抑制炎症因子的活性成分抗动脉粥样硬化研究进展[J].中南药学,2013,11(4):289-292.
[15] 陈芬燕,郭韧,张毕奎.丹参酮ⅡA的心血管药理作用研究进展[J].中国中药杂志,2015,40(9):1649-1653.
[16] Chen J,Shi D Y,Liu S L,et al.Tanshinone ⅡA induces growth inhibition and apoptosis in gastric cancer in vitro and in vivo[J].Oncol Rep,2012,27:523-528.
[17] 赵丕文,牛建昭,王继峰,等.丹参酮ⅡA抗乳腺癌细胞增殖作用研究[J].中国药理学通报,2010,26(7):903-906.
[18] Liu C,Li J,Wang L,et al.Analysis of tanshinone ⅡA induced cellular apoptosis in leukemia cells by genome-wide expression profiling[J].BMC Complement Altern Med,2012,12:5.
[19] Shan Y F,Shen X,Xie Y K,et al.Inhibitory effects of tanshinone Ⅱ-A on invasion and metastasis of human colon carcinoma cells[J].Acta Pharmacol Sin,2009,30(11):1537-1542.
[20] 中国科学院西北高原生物研究所·青海植物志委员会.青海植物志[M].西宁:青海人民出版社,1996:153.
[21] 程沛,韩东岐,胡伟慧,等.高效液相色谱法同时测定丹参中10种水溶性和4种脂溶性成分的含量[J].药物分析杂志,2015,35(6):991-996.
[22] 胡伟慧,韩东岐,江坤,等.RP-HPLC法同时测定甘西鼠尾草中12种成分的含量[J].药物分析杂志,2016,36(3):486-493.
[2] 王艳伟,代雪平,王晓伟.HPLC法测定益心通脉颗粒中丹参酮Ⅰ、隐丹参酮和丹参酮ⅡA的含量[J].西北药学杂志,2018,33(4):471-474.
[3] 陆丽萍.甘西鼠尾草化学成分及药理作用研究进展[J].中医临床研究,2016,20(8):140-141.
[4] 杨阳,张凤,蔡飞,等.甘西鼠尾草化学成分及药理作用研究进展[J].中药材,2008,31(5):787-790.
[5] 中国科学院中国植物志编辑委员会.中国植物志:66卷[M].北京:科学出版社,1977:86.
[6] 林超,刘兆国,钱星,等.丹酚酸B在心血管疾病中药理作用研究进展[J].中国药理学通报,2015,31(4):449-452.
[7] 陈俞材,方莲花,杜冠华,等.丹参水溶性化合物抗心肌缺血作用的研究进展[J].中国药理学通报,2015,31(2):162-165.
[8] 罗慧英,程体娟.甘西鼠尾草注射液与丹参注射液对化学性缺氧小鼠能量代谢影响的比较[J].中国临床药理学与治疗学,2004,9(2):174-176.
[9] 程体娟,罗慧英,王巧明,等.甘西鼠尾草注射液与丹参注射液对大鼠皮层神经细胞缺血性损伤保护作用的比较研究[J].中国药理与临床,2005,21(3):34-36.
[10] 程体娟,杨志勇.甘西鼠尾草注射液和丹参注射液抗大鼠急性脑缺血和抗脂质过氧化作用的比较[J].中国临床药理学与治疗学,2003,8(1):23-26.
[11] 叶剑.丹参的药用成分与药理作用探析[J].陕西中医学院学报,2012,35(5):71-73.
[12] 杨阳,朱斌,吴志军,等.甘西鼠尾草提取物对正常大鼠全血粘度影响及对水负荷大鼠利尿作用[J].中国医院药学杂志,2012,32(10):751-754.
[13] 林超,刘兆国,钱星,等.丹酚酸B在心血管疾病中药理作用研究进展[J].中国药理学通报,2015,31(4):449-452.
[14] 杨阳,王林,张毕奎.丹参中抑制炎症因子的活性成分抗动脉粥样硬化研究进展[J].中南药学,2013,11(4):289-292.
[15] 陈芬燕,郭韧,张毕奎.丹参酮ⅡA的心血管药理作用研究进展[J].中国中药杂志,2015,40(9):1649-1653.
[16] Chen J,Shi D Y,Liu S L,et al.Tanshinone ⅡA induces growth inhibition and apoptosis in gastric cancer in vitro and in vivo[J].Oncol Rep,2012,27:523-528.
[17] 赵丕文,牛建昭,王继峰,等.丹参酮ⅡA抗乳腺癌细胞增殖作用研究[J].中国药理学通报,2010,26(7):903-906.
[18] Liu C,Li J,Wang L,et al.Analysis of tanshinone ⅡA induced cellular apoptosis in leukemia cells by genome-wide expression profiling[J].BMC Complement Altern Med,2012,12:5.
[19] Shan Y F,Shen X,Xie Y K,et al.Inhibitory effects of tanshinone Ⅱ-A on invasion and metastasis of human colon carcinoma cells[J].Acta Pharmacol Sin,2009,30(11):1537-1542.
[20] 中国科学院西北高原生物研究所·青海植物志委员会.青海植物志[M].西宁:青海人民出版社,1996:153.
[21] 程沛,韩东岐,胡伟慧,等.高效液相色谱法同时测定丹参中10种水溶性和4种脂溶性成分的含量[J].药物分析杂志,2015,35(6):991-996.
[22] 胡伟慧,韩东岐,江坤,等.RP-HPLC法同时测定甘西鼠尾草中12种成分的含量[J].药物分析杂志,2016,36(3):486-493.