黔产钩藤药材茎枝和叶的HPLC指纹图谱比较
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摘要
目的:比较黔产钩藤带钩茎枝和叶的HPLC指纹图谱,分析二者化学成分的差异,为钩藤叶的后续研究提供实验依据。方法:采用《中药色谱指纹图谱相似度评价系统》2004A版计算钩藤带钩茎枝和叶各10批样品的HPLC指纹图谱的相似度,以钩藤碱和异钩藤碱为指标成分进行指纹图谱比较,色谱条件为Kromasil C18色谱柱(4.6 mm×250 mm,5μm),流动相甲醇(A)-0.02%氨水(B)梯度洗脱(0~45 min,30%~60%A;45~75 min,60%~64%A;75~90 min,64%~70%A;90~95 min,70%~70%A),流速1.0 mL·min-1,柱温30℃,检测波长245 nm,进样量10μL。结果:确定了16个共有峰构成10批钩藤茎枝药材的特征指纹图谱,各批样品的相似度均>0.87;16个共有峰构成10批钩藤叶特征指纹图谱,各批样品的相似度均>0.80。结论:钩藤叶中异钩藤碱含量高于钩藤茎枝,但二者含有的钩藤碱含量相差不大。建立的指纹图谱条件能用于比对钩藤茎枝和叶中主要化学成分,为钩藤叶的药用研究提供参考。
Objective: To establish HPLC fingerprint for comparative studying on chemical substances difference between stems and leaves of Uncariae Ramulus Cum Uncis,in order to lay scientific basis for follow-up studies of Uncariae Ramulus Cum Uncis leaves. Method: Similarity of HPLC fingerprint was calculated by 2004 A version of "traditional medicine fingerprint similarity software evaluation system"for 10 batches of stems and 10 batches of leaves in Uncariae Ramulus Cum Uncis. Taking rhynchophylline and isorhynchophylline as index components,HPLC analysis was performed on a Kromasil column( 4. 6 mm × 250 mm,5 μm) with gradient eluting system consisted of methanol( A)-0. 02% ammonia( B)( 0-45 min,30%-60% A; 45-75 min,60%-64% A; 75-90 min,64%-70% A; 90-95 min,70%-70% A), flow rate was 1. 0 mL ·min- 1,column temperature was maintained at 30 ℃ and detection wavelength was set at 245 nm,injection volume of 10 μL.Result: Sixteen common peaks were obtained in 10 batches of Uncariae Ramulus Cum Uncis stems with similarity higher than 0. 87 in each batches of samples; sixteen common peaks were obtained in 10 batches of Uncariae Ramulus Cum Uncis leaves with similarity higher than 0. 80 in each batches of samples. Conclusion: The content of isorhynchophylline in leaves is higher than that in leaves, but the content of rhynchophylline have little difference. This established fingerprint conditions can be used as initial comparative studies on main chemical components of Uncariae Ramulus Cum Uncis stems and leaves,it provide a reference for medicinal research of Uncariae Ramulus Cum Uncis leaves.
Objective: To establish HPLC fingerprint for comparative studying on chemical substances difference between stems and leaves of Uncariae Ramulus Cum Uncis,in order to lay scientific basis for follow-up studies of Uncariae Ramulus Cum Uncis leaves. Method: Similarity of HPLC fingerprint was calculated by 2004 A version of "traditional medicine fingerprint similarity software evaluation system"for 10 batches of stems and 10 batches of leaves in Uncariae Ramulus Cum Uncis. Taking rhynchophylline and isorhynchophylline as index components,HPLC analysis was performed on a Kromasil column( 4. 6 mm × 250 mm,5 μm) with gradient eluting system consisted of methanol( A)-0. 02% ammonia( B)( 0-45 min,30%-60% A; 45-75 min,60%-64% A; 75-90 min,64%-70% A; 90-95 min,70%-70% A), flow rate was 1. 0 mL ·min- 1,column temperature was maintained at 30 ℃ and detection wavelength was set at 245 nm,injection volume of 10 μL.Result: Sixteen common peaks were obtained in 10 batches of Uncariae Ramulus Cum Uncis stems with similarity higher than 0. 87 in each batches of samples; sixteen common peaks were obtained in 10 batches of Uncariae Ramulus Cum Uncis leaves with similarity higher than 0. 80 in each batches of samples. Conclusion: The content of isorhynchophylline in leaves is higher than that in leaves, but the content of rhynchophylline have little difference. This established fingerprint conditions can be used as initial comparative studies on main chemical components of Uncariae Ramulus Cum Uncis stems and leaves,it provide a reference for medicinal research of Uncariae Ramulus Cum Uncis leaves.
引文
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[2]莫志贤,许丹丹.钩藤临床运用的新进展[J].时珍国医国药,2006,17(5):684.
[3]霍青,赵婧,李运伦,等.钩藤临床应用研究概况[J].山东中医药杂志,2010,29(6):426.
[4]汪冰.新药用资源钩藤叶的基础研究[D].沈阳:沈阳药科大学,2006.
[5]张峻,杨成金,吴大刚,等.钩藤的化学成分研究[J].中草药,1998,29(10):649.
[6]叶齐,齐荔红.钩藤的主要成分及生物活性研究进展[J].西北药学杂志,2012,27(5):508.
[7]林晓亮.毛钩藤叶的生药学鉴定及药理学研究[D].广州:南方医科大学,2007.
[8]胡岚岚,汤建林.HPLC-ELSD测定中药钩藤中钩藤碱的含量[J].重庆医学,2013,42(10):1089.
[9]杨秀娟,洪燕龙,吴飞,等.HPLC测定钩藤中和异钩藤碱的方法学探讨[J].中国中药杂志,2013,38(5):720.
[10]广西壮族自治区革命委员会卫生局.广西本草选编.上册[M].桂林:广西人民出版社,1974:886.