HPLC法同时测定活血促愈胶囊中2种黄酮类和4种菲醌类成分的含量
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摘要
目的:建立同时测定活血促愈胶囊中2种黄酮类(芦丁、山柰酚-3-O-芸香糖苷)和4种菲醌类成分(二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ、丹参酮Ⅱ_A)含量的方法。方法:采用高效液相色谱法。色谱柱为Welch Ultimate XB-C18,流动相为乙腈-0.1%磷酸溶液(梯度洗脱),流速为1.0 mL/min,柱温为20℃,检测波长为270 nm,进样量为10μL。结果:芦丁、山柰酚-3-O-芸香糖苷、二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ、丹参酮Ⅱ_A的检测质量浓度线性范围分别为172.13～860.66μg/mL(r=0.999 7)、15.33～76.66μg/mL(r=0.999 8)、12.81～64.06μg/mL(r=0.999 3)、5.90～29.52μg/m L(r=0.999 3)、5.12～25.60μg/mL(r=0.999 2)、6.71～33.57μg/m L(r=0.999 7),检测限分别为0.08、0.01、0.01、0.01、0.01、0.01μg/mL,定量限分别为0.27、0.02、0.03、0.03、0.03、0.03μg/m L;精密度、稳定性、重复性试验的RSD均小于2.0%(n=6);加样回收率分别为97.54%～100.25%(RSD=1.07%,n=6)、96.90%～101.91%(RSD=1.73%,n=6)、96.24%～102.89%(RSD=2.32%,n=6)、97.04%～102.18%(RSD=1.82%,n=6)、95.06%～97.73%(RSD=1.18%,n=6)、95.59%～101.40%(RSD=2.29%,n=6)。结论:该方法灵敏、快速、操作简便、重复性好,可用于活血促愈胶囊中芦丁、山柰酚-3-O-芸香糖苷、二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ、丹参酮Ⅱ_A含量的同时测定。
OBJECTIVE:To establish a method for simultaneous determination of two flavonoids(rutin and kaempferol-3-Orutinoside) and four phenoquinones(dihydrotanshinone Ⅰ,cryptotanshinone,tanshinone Ⅰ and tanshinone Ⅱ_A)in Huoxue cuyu capsules. METHODS:HPLC method was adopted. The determination was performed on Welch Ultimate XB-C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution(gradient elution)at the flow rate of 1.0 mL/min. The column temperature was set at 20 ℃,and detection wavelength was set at 270 nm. The sample size was 10 μL. RESULTS:The linear range of rutin,kaempferol-3-O-rutinoside,dihydrotanshinone Ⅰ,cryptotanshinone,tanshinone Ⅰ and tanshinoneⅡ_Awere 172.13-860.66 μg/m L(r=0.999 7),15.33-76.66 μg/mL(r=0.999 8),12.81-64.06 μg/mL(r=0.999 3),5.90-29.52 μg/mL(r=0.999 3),5.12-25.60 μg/m L(r=0.999 2),6.71-33.57 μg/mL(r=0.999 7),respectively. The limits of detection were 0.08,0.01,0.01,0.01,0.01,0.01 μg/mL. The limits of quantitation were 0.27,0.02,0.03,0.03,0.03,0.03 μg/mL,respectively. RSDs of precision,stability test and repetition tests were all lower than 2.0%(n=6). The recoveries were 97.54%-100.25%(RSD=1.07%,n=6),96.90%-101.91%(RSD=1.73%,n=6),96.24%-102.89%(RSD=2.32%,n=6),97.04%-102.18%(RSD=1.82%,n=6),95.06%-97.73%(RSD=1.18%,n=6),95.59%-101.40%(RSD=2.29%,n=6),respectively. CONCLUSIONS:The method is sensitive,rapid,simple and has good reproducibility. It can be used for simultaneous determination of rutin, kaempferol-3-O-rutinoside, dihydrotanshinone Ⅰ,cryptotanshinone,tanshinone Ⅰ,tanshinoneⅡ_A in Huoxue cuyu capsules.
OBJECTIVE:To establish a method for simultaneous determination of two flavonoids(rutin and kaempferol-3-Orutinoside) and four phenoquinones(dihydrotanshinone Ⅰ,cryptotanshinone,tanshinone Ⅰ and tanshinone Ⅱ_A)in Huoxue cuyu capsules. METHODS:HPLC method was adopted. The determination was performed on Welch Ultimate XB-C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution(gradient elution)at the flow rate of 1.0 mL/min. The column temperature was set at 20 ℃,and detection wavelength was set at 270 nm. The sample size was 10 μL. RESULTS:The linear range of rutin,kaempferol-3-O-rutinoside,dihydrotanshinone Ⅰ,cryptotanshinone,tanshinone Ⅰ and tanshinoneⅡ_Awere 172.13-860.66 μg/m L(r=0.999 7),15.33-76.66 μg/mL(r=0.999 8),12.81-64.06 μg/mL(r=0.999 3),5.90-29.52 μg/mL(r=0.999 3),5.12-25.60 μg/m L(r=0.999 2),6.71-33.57 μg/mL(r=0.999 7),respectively. The limits of detection were 0.08,0.01,0.01,0.01,0.01,0.01 μg/mL. The limits of quantitation were 0.27,0.02,0.03,0.03,0.03,0.03 μg/mL,respectively. RSDs of precision,stability test and repetition tests were all lower than 2.0%(n=6). The recoveries were 97.54%-100.25%(RSD=1.07%,n=6),96.90%-101.91%(RSD=1.73%,n=6),96.24%-102.89%(RSD=2.32%,n=6),97.04%-102.18%(RSD=1.82%,n=6),95.06%-97.73%(RSD=1.18%,n=6),95.59%-101.40%(RSD=2.29%,n=6),respectively. CONCLUSIONS:The method is sensitive,rapid,simple and has good reproducibility. It can be used for simultaneous determination of rutin, kaempferol-3-O-rutinoside, dihydrotanshinone Ⅰ,cryptotanshinone,tanshinone Ⅰ,tanshinoneⅡ_A in Huoxue cuyu capsules.
引文
[1]徐琳本,陈丽萍,肖梅英.活血促愈胶囊对外伤血瘀证大鼠模型的影响[J].中国实验方剂学杂志,2013,19(4):270-273.
[2]邹旦,彭六明,肖放军,等.活血促愈胶囊治疗骨折早期软组织损伤30例[J].湖南中医杂志,2012,28(2):32-33.
[3]陈丹,李柯,卢茂芳,等. HPLC法同时测定活血促愈胶囊中5个主要活性成分的含量[J].药物分析杂志,2017,37(12):2185-2190.
[4]王笑,王雨,张冰,等.槐不同药用部位本草学、化学成分和药理作用研究进展[J].中草药,2018,49(18):4461-4467.
[5]国家药典委员会.中华人民共和国药典:一部[S].2015年版.北京:中国医药科技出版社,2015:76、354.
[6]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.
[7]张慧娴,杜守颖,陆洋,等. HPLC同时测定丹参水溶性及脂溶性5种成分的含量[J].中国实验方剂学杂志,2010,16(15):34-38.
[8]舒文将,刘金磊,邹蓉,等.高效液相色谱法测定不同产地和采收期对槐米中芦丁含量的影响[J].时珍国医国药,2017,28(3):709-711.
[9]李俊,王冬,张丽梅,等.复方黄槐片质量标准的提高[J].中国药师,2017,20(12):2251-2254.
[10]程中琴,刘小妹,施崇精,等. HPLC法同时测定平脏调神颗粒中8种成分的含量[J].中国药房,2018,29(1):33-37.
[11]付娟,张海弢,杨素德,等. HPLC法同时测定芪白平肺颗粒中4种皂苷类成分的含量[J].中国药房,2015,26(33):4698-4700.
[12]王姗,张毅,张建华,等. HPLC法同时测定金钱草药材中芦丁和烟花苷的含量[J].沈阳药科大学学报,2017,34(12):1067-1071.
[13]符诗聪,史炜镔,张昊,等.丹参有效部位(丹参-9403)对骨折愈合的骨计量学研究[J].中国中西医结合杂志,2000(S1):116-117.
[14]赵仁霞.丹参的现代药理研究及临床应用[J].中国医药指南,2011,9(12):291-292.
[15]戴新新,宿树兰,郭盛,等.丹参酮类成分的生物活性与应用开发研究进展[J].中草药,2017,48(7):1442-1448.
[16]张文通,李俊,吴玉婷,等.山柰酚-3-O-芸香糖苷对血管平滑肌细胞增殖、迁移及TGFBR1信号通路活化的影响[J].中国病理生理杂志,2018,34(5):832-838.
[17]李秋红,栾仲秋,王继坤.中药槐米的化学成分、炮制研究及药理作用研究进展[J].中医药学报,2017,45(3):112-116.
[18]方小燕.聚酰胺树脂在分离纯化黄酮中的应用研究[J].海峡药学,2013,25(5):41-42.
[19]李梓盟,张柯达,吴金虎.聚酰胺树脂纯化复方“前愈”总黄酮的工艺研究[J].中国新药杂志,2016,25(8):913-916.
[2]邹旦,彭六明,肖放军,等.活血促愈胶囊治疗骨折早期软组织损伤30例[J].湖南中医杂志,2012,28(2):32-33.
[3]陈丹,李柯,卢茂芳,等. HPLC法同时测定活血促愈胶囊中5个主要活性成分的含量[J].药物分析杂志,2017,37(12):2185-2190.
[4]王笑,王雨,张冰,等.槐不同药用部位本草学、化学成分和药理作用研究进展[J].中草药,2018,49(18):4461-4467.
[5]国家药典委员会.中华人民共和国药典:一部[S].2015年版.北京:中国医药科技出版社,2015:76、354.
[6]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.
[7]张慧娴,杜守颖,陆洋,等. HPLC同时测定丹参水溶性及脂溶性5种成分的含量[J].中国实验方剂学杂志,2010,16(15):34-38.
[8]舒文将,刘金磊,邹蓉,等.高效液相色谱法测定不同产地和采收期对槐米中芦丁含量的影响[J].时珍国医国药,2017,28(3):709-711.
[9]李俊,王冬,张丽梅,等.复方黄槐片质量标准的提高[J].中国药师,2017,20(12):2251-2254.
[10]程中琴,刘小妹,施崇精,等. HPLC法同时测定平脏调神颗粒中8种成分的含量[J].中国药房,2018,29(1):33-37.
[11]付娟,张海弢,杨素德,等. HPLC法同时测定芪白平肺颗粒中4种皂苷类成分的含量[J].中国药房,2015,26(33):4698-4700.
[12]王姗,张毅,张建华,等. HPLC法同时测定金钱草药材中芦丁和烟花苷的含量[J].沈阳药科大学学报,2017,34(12):1067-1071.
[13]符诗聪,史炜镔,张昊,等.丹参有效部位(丹参-9403)对骨折愈合的骨计量学研究[J].中国中西医结合杂志,2000(S1):116-117.
[14]赵仁霞.丹参的现代药理研究及临床应用[J].中国医药指南,2011,9(12):291-292.
[15]戴新新,宿树兰,郭盛,等.丹参酮类成分的生物活性与应用开发研究进展[J].中草药,2017,48(7):1442-1448.
[16]张文通,李俊,吴玉婷,等.山柰酚-3-O-芸香糖苷对血管平滑肌细胞增殖、迁移及TGFBR1信号通路活化的影响[J].中国病理生理杂志,2018,34(5):832-838.
[17]李秋红,栾仲秋,王继坤.中药槐米的化学成分、炮制研究及药理作用研究进展[J].中医药学报,2017,45(3):112-116.
[18]方小燕.聚酰胺树脂在分离纯化黄酮中的应用研究[J].海峡药学,2013,25(5):41-42.
[19]李梓盟,张柯达,吴金虎.聚酰胺树脂纯化复方“前愈”总黄酮的工艺研究[J].中国新药杂志,2016,25(8):913-916.