南板蓝根质量评价研究及马蓝愈伤组织培养研究
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摘要
南板蓝根为爵床科植物马蓝Baphicacanthus cusia (Nees) Bremek的干燥根茎及根。具有清热解毒、凉血的功效。广泛用于病毒性传染病,如乙型脑炎、流行性腮腺炎、急性传染性肝炎、感冒(流感)和非典型性肺炎。现今《中国药典》南板蓝根质量标准尚不完善,而商品药材入药部位与各级质量标准均不吻合。
本文在《中国药典》2005年版的基础上,首次建立了HPLC测定南板蓝根药材中主要指标成分靛玉红和靛蓝含量的方法,对全国南板蓝根主产地5省的22份样品进行了质量评价研究,完善了药典指标性成分的定量分析。建议南板蓝根药材按干燥品计算,含靛玉红(C16H1oN202)不得少于0.010%,含靛蓝(C16H10N2O2)不得少于0.015%。
四川宜宾南溪县大坪乡产根茎、根及茎中靛玉红和靛蓝含量均高于其它产地相同部位样品,靛玉红的含量可达0.087%,靛蓝可达0.136%。对马蓝叶中靛玉红、靛蓝含量测定表明,叶的含量明显高于根及根茎和茎,四川都江堰产马蓝叶中靛玉红和靛蓝含量最高,靛玉红的含量可达0.949%,靛蓝可达0.361%。
首次进行了不同产地南板蓝根药材HPLC指纹图谱研究,各产地南板蓝根药材与对照色谱指纹图谱的相似度较好。马蓝不同药用部位的指纹图谱的相关性分析结果显示,茎与根茎及根的相似度较好,而叶与其它部位差异较大。
根据南板蓝根凝集素与抑制流感病毒的效果成正比关系,首次建立了南板蓝根植物凝集素提取和测定方法,不同产地南板蓝根药材中凝集素含量差异较大。马蓝的茎与根茎及根所含凝集素相当,而同一产地样品叶中凝集素水平较低。血细胞凝集活性最强的为广西防城十万山产马蓝根茎及根、另外广西横县产马蓝茎、广东梅县平远产马蓝茎、四川宜宾筠连产马蓝茎凝集活性亦较强。
首次对南板蓝根商品药材入药部位进行了调查,结果表明商品药材主要入药部位为茎基部,有少部分商品药材带有少量的根茎及根,未见纯粹以根茎及根入药的商品药材,与《中国药典》2005年版来源项规定不相符。本文首次以指标性成分的含量测定结合化学指纹图谱研究、生物测定研究,结果表明马蓝的茎与根茎及根可以作为同一药材使用。
首次测定了南板蓝根药材及马蓝其它药用部位中重金属和有害元素含量。南板蓝根中As、Hg的含量均低于《药用植物及制剂外经贸绿色行业标准》的规定,但部分样品Pb、Cd、Cu的含量超标,其中以Cd超标情况最为严重。地上茎做药用的南板蓝根少有重金属超标的情况,马蓝叶中Pb超标情况较根和茎部位严重,可能原因是植株吸收土壤中的铅,富集于叶器官中。
首次研究了不同激素对南板蓝根叶片、茎段愈伤组织诱导的影响,确定了马蓝愈伤组织诱导培养基：MS+6-BA1.0mg／l+NAA0.1mg／l。测定了愈伤组织中靛蓝、靛玉红含量,结果愈伤组织含量为原植株茎部位的8倍,其抗病毒活性的相对凝集素含量为原植株的4倍。
以上工作为综合评价南板蓝根药材质量,并为南板蓝根新资源的开发和保护提供了科学依据。
Baphicacanthus cusia (Nees) Bremek.is the dry rhizome and root of Baphicacanthus cusia (Nees) Bremek. It has cold nature, bitter flavor and enters Heart and Stomach channels. The actions of Baphicacanthus cusia (Nees) Bremek. is clearing heat and detoxification and cooling blood. It's widely applied for epidemic virus control, such as epidemic encephalitis B, epidemic parotitis, acute epidemic hepatitis, common cold (influenza) and SARS
The recent herbal quality standards for Baphicacanthus cusia (Nees) Bremek. is not completed and most of them are still using the above ground stem as Baphicacanthus cusia (Nees) Bremek.. In this article, we established the HPLC tests for Baphicacanthus cusia (Nees) Bremek. samples from different source regions and examination on indigo and Bindirubin contents, which are considered as the index components from different herbal parts of Baphicacanthus cusia (Nees) Bremek. plant. Comparing with recent statistics, this research provides more completed and reliable quality evaluation and quality control standards for Baphicacanthus cusia (Nees) Bremek. The result indicated that the Bindirubin (C16H10N2O2) content should not be less than 0.010%, while indigo(C16H10N2O2)content should be not less than 0.015%, according to the Baphicacanthus cusia (Nees) Bremek. samples from various source regions.
This article also provides the fingerprint analysis for Baphicacanthus cusia (Nees) Bremek.samples from various source region. The fingerprint between the samples and the control chromatographic has good similarities. The relational analysis of fingerprint on different herbal parts of Baphicacanthus cusia (Nees) Bremek. shows that the rhizome and root have better similarities, comparing with the leaf and other parts.
The methods of extracting and testing lecin in Baphicacanthus cusia (Nees) Bremek. have been established, according to the direct ratio between Baphicacanthus cusia (Nees) Bremek. lecin and its influenza virus inhibition effects. The Baphicacanthus cusia (Nees) Bremek. from different souse regions shows obvious difference on lecin content. The Northern Baphicacanthus cusia (Nees) Bremek. has no obvious blood coagulation effects. When we test the same source region samples, the rhizome, root and stem have equal lecin, but the leaf has lower lecin level.
As, Hg contents in Baphicacanthus cusia (Nees) Bremek. is under the limitation in Foreign Trade and Economic Green Industry Criterion of Medicinal Plants and Pharmaceutics. But some samples has Pb、Cd、Cu contents above quota, especially Cd. The upper ground stem rarely has heavy metal above quota. Pb above quota is severer in leaf than in rhizome and the reason could be that the plant absorbed Pb from soil and Pb accumulated in leaves.
To compare the results from bioavailability tests and chemical components analysis on Baphicacanthus cusia (Nees) Bremek. from different souse region, we found that contents of Bindirubin and indigo have positive correlation and direct ratio with the content of lecin from rhizome, root and stem have direct relational in all samples. But negative correlation results present in the leaf samples. Combining these results with fingerprint research, we can give the conclusion that the rhizome, root and stem of Baphicacanthus cusia (Nees) Bremek. can be applied as one herb. Since its leaf has significant differences in chemical components and anti virus bioavailability test results, Baphicacanthus cusia (Nees) Bremek. should be used as a separate herbal part.
We researched different hormone influences on Baphicacanthus cusia (Nees) Bremek. leaf and stem inducement callus and ensured the Baphicacanthus cusia (Nees) Bremek. callus culture medium:MS+6-BA1.0mg/l+NAA0.1mg/l. The indigo, Bindirubincontentis 8 times as former stem parts of the plants and the lecin relative content of anti virus ingredient reaches 4 times as that of before.
本文在《中国药典》2005年版的基础上,首次建立了HPLC测定南板蓝根药材中主要指标成分靛玉红和靛蓝含量的方法,对全国南板蓝根主产地5省的22份样品进行了质量评价研究,完善了药典指标性成分的定量分析。建议南板蓝根药材按干燥品计算,含靛玉红(C16H1oN202)不得少于0.010%,含靛蓝(C16H10N2O2)不得少于0.015%。
四川宜宾南溪县大坪乡产根茎、根及茎中靛玉红和靛蓝含量均高于其它产地相同部位样品,靛玉红的含量可达0.087%,靛蓝可达0.136%。对马蓝叶中靛玉红、靛蓝含量测定表明,叶的含量明显高于根及根茎和茎,四川都江堰产马蓝叶中靛玉红和靛蓝含量最高,靛玉红的含量可达0.949%,靛蓝可达0.361%。
首次进行了不同产地南板蓝根药材HPLC指纹图谱研究,各产地南板蓝根药材与对照色谱指纹图谱的相似度较好。马蓝不同药用部位的指纹图谱的相关性分析结果显示,茎与根茎及根的相似度较好,而叶与其它部位差异较大。
根据南板蓝根凝集素与抑制流感病毒的效果成正比关系,首次建立了南板蓝根植物凝集素提取和测定方法,不同产地南板蓝根药材中凝集素含量差异较大。马蓝的茎与根茎及根所含凝集素相当,而同一产地样品叶中凝集素水平较低。血细胞凝集活性最强的为广西防城十万山产马蓝根茎及根、另外广西横县产马蓝茎、广东梅县平远产马蓝茎、四川宜宾筠连产马蓝茎凝集活性亦较强。
首次对南板蓝根商品药材入药部位进行了调查,结果表明商品药材主要入药部位为茎基部,有少部分商品药材带有少量的根茎及根,未见纯粹以根茎及根入药的商品药材,与《中国药典》2005年版来源项规定不相符。本文首次以指标性成分的含量测定结合化学指纹图谱研究、生物测定研究,结果表明马蓝的茎与根茎及根可以作为同一药材使用。
首次测定了南板蓝根药材及马蓝其它药用部位中重金属和有害元素含量。南板蓝根中As、Hg的含量均低于《药用植物及制剂外经贸绿色行业标准》的规定,但部分样品Pb、Cd、Cu的含量超标,其中以Cd超标情况最为严重。地上茎做药用的南板蓝根少有重金属超标的情况,马蓝叶中Pb超标情况较根和茎部位严重,可能原因是植株吸收土壤中的铅,富集于叶器官中。
首次研究了不同激素对南板蓝根叶片、茎段愈伤组织诱导的影响,确定了马蓝愈伤组织诱导培养基：MS+6-BA1.0mg／l+NAA0.1mg／l。测定了愈伤组织中靛蓝、靛玉红含量,结果愈伤组织含量为原植株茎部位的8倍,其抗病毒活性的相对凝集素含量为原植株的4倍。
以上工作为综合评价南板蓝根药材质量,并为南板蓝根新资源的开发和保护提供了科学依据。
Baphicacanthus cusia (Nees) Bremek.is the dry rhizome and root of Baphicacanthus cusia (Nees) Bremek. It has cold nature, bitter flavor and enters Heart and Stomach channels. The actions of Baphicacanthus cusia (Nees) Bremek. is clearing heat and detoxification and cooling blood. It's widely applied for epidemic virus control, such as epidemic encephalitis B, epidemic parotitis, acute epidemic hepatitis, common cold (influenza) and SARS
The recent herbal quality standards for Baphicacanthus cusia (Nees) Bremek. is not completed and most of them are still using the above ground stem as Baphicacanthus cusia (Nees) Bremek.. In this article, we established the HPLC tests for Baphicacanthus cusia (Nees) Bremek. samples from different source regions and examination on indigo and Bindirubin contents, which are considered as the index components from different herbal parts of Baphicacanthus cusia (Nees) Bremek. plant. Comparing with recent statistics, this research provides more completed and reliable quality evaluation and quality control standards for Baphicacanthus cusia (Nees) Bremek. The result indicated that the Bindirubin (C16H10N2O2) content should not be less than 0.010%, while indigo(C16H10N2O2)content should be not less than 0.015%, according to the Baphicacanthus cusia (Nees) Bremek. samples from various source regions.
This article also provides the fingerprint analysis for Baphicacanthus cusia (Nees) Bremek.samples from various source region. The fingerprint between the samples and the control chromatographic has good similarities. The relational analysis of fingerprint on different herbal parts of Baphicacanthus cusia (Nees) Bremek. shows that the rhizome and root have better similarities, comparing with the leaf and other parts.
The methods of extracting and testing lecin in Baphicacanthus cusia (Nees) Bremek. have been established, according to the direct ratio between Baphicacanthus cusia (Nees) Bremek. lecin and its influenza virus inhibition effects. The Baphicacanthus cusia (Nees) Bremek. from different souse regions shows obvious difference on lecin content. The Northern Baphicacanthus cusia (Nees) Bremek. has no obvious blood coagulation effects. When we test the same source region samples, the rhizome, root and stem have equal lecin, but the leaf has lower lecin level.
As, Hg contents in Baphicacanthus cusia (Nees) Bremek. is under the limitation in Foreign Trade and Economic Green Industry Criterion of Medicinal Plants and Pharmaceutics. But some samples has Pb、Cd、Cu contents above quota, especially Cd. The upper ground stem rarely has heavy metal above quota. Pb above quota is severer in leaf than in rhizome and the reason could be that the plant absorbed Pb from soil and Pb accumulated in leaves.
To compare the results from bioavailability tests and chemical components analysis on Baphicacanthus cusia (Nees) Bremek. from different souse region, we found that contents of Bindirubin and indigo have positive correlation and direct ratio with the content of lecin from rhizome, root and stem have direct relational in all samples. But negative correlation results present in the leaf samples. Combining these results with fingerprint research, we can give the conclusion that the rhizome, root and stem of Baphicacanthus cusia (Nees) Bremek. can be applied as one herb. Since its leaf has significant differences in chemical components and anti virus bioavailability test results, Baphicacanthus cusia (Nees) Bremek. should be used as a separate herbal part.
We researched different hormone influences on Baphicacanthus cusia (Nees) Bremek. leaf and stem inducement callus and ensured the Baphicacanthus cusia (Nees) Bremek. callus culture medium:MS+6-BA1.0mg/l+NAA0.1mg/l. The indigo, Bindirubincontentis 8 times as former stem parts of the plants and the lecin relative content of anti virus ingredient reaches 4 times as that of before.
引文
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