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2021, (14) 3514-3521

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“失传”中药材宣黄连的分子鉴定与生物碱含量分析
Molecular identification and alkaloids content of endangered Chinese medicinal plant Coptis chinensis var. brevisepala

王安琪;李西文;袁庆军;郭宁;杨滨;孙奕;
WANG An-qi;LI Xi-wen;YUAN Qing-jun;GUO Ning;YANG Bin;SUN Yi;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences;National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;

摘要(Abstract):

据本草记载,宣黄连为古代医家推崇的道地药材。与其他黄连品种相比,其根茎“节如连珠”,具有很高的药用价值。然而,宣黄连作为历史“失传”中药材,它的药效物质与药用价值尚未得到深入阐明。因此,该研究基于液质联用技术对宣黄连根茎中药效成分进行了定量测定,比较分析了宣黄连与味连等3个《中国药典》黄连品种的异喹啉生物碱含量差异;采用分子鉴定技术准确地鉴定出宣黄连12批,味连9批,雅连4批和云连1批。选用Waters CORTECS C18色谱柱(4.6 mm×150 mm,2.7μm),以乙腈-0.4%甲酸水为流动相进行梯度洗脱。质谱分析采用电喷雾正离子模式采集,定量结果显示8个主要生物碱成分在考察的浓度范围内线性关系良好(R2>0.996),回收率为95.18%2>0.996),回收率为95.18%105.0%,RSD为0.28%105.0%,RSD为0.28%3.7%。宣黄连根茎与其他种黄连根茎相比,小檗碱和非洲防己碱含量最高,且生物碱总量与味连相近而高于雅连和云连;宣黄连的根茎中生物碱含量高于根和地上部分。应用主成分分析(PCA)可区分出不同品种黄连的主要生物碱成分。研究结果表明,该分析方法可靠、高效,为宣黄连的质量研究提供参考。
According to the records of Chinese materia medica,Coptis chinensis var. brevisepala is an authentic Chinese medicinal plant highly recommended by ancient physicians since its rhizome is like a string of beads and has a good medicinal value. However,its medicinal components and values remain to be studied as it is endangered because of overexploitation. Therefore,this study aims to quantitatively determine its effective components based on UPLC-QTOF-MS,and to compare the contents of isoquinoline alkaloids in C.chinensis var. brevisepala with those in other Coptis species. Meanwhile,molecular methods accurately identified 12 batches of C. chinensis var. brevisepala,9 batches of C. chinensis,4 batches of C. deltoidea,and 1 batch of C. teeta. Gradient elution was performed with Waters CORTECS C18 column( 4. 6 mm× 150 mm,2. 7 μm) and the mobile phase acetonitrile-water with 0. 4% formic acid. Mass spectrometry was conducted in ESI positive mode. The quantitative results showed that 8 main alkaloids had a good linear relationship within the concentration range( R3.7%。宣黄连根茎与其他种黄连根茎相比,小檗碱和非洲防己碱含量最高,且生物碱总量与味连相近而高于雅连和云连;宣黄连的根茎中生物碱含量高于根和地上部分。应用主成分分析(PCA)可区分出不同品种黄连的主要生物碱成分。研究结果表明,该分析方法可靠、高效,为宣黄连的质量研究提供参考。
According to the records of Chinese materia medica,Coptis chinensis var. brevisepala is an authentic Chinese medicinal plant highly recommended by ancient physicians since its rhizome is like a string of beads and has a good medicinal value. However,its medicinal components and values remain to be studied as it is endangered because of overexploitation. Therefore,this study aims to quantitatively determine its effective components based on UPLC-QTOF-MS,and to compare the contents of isoquinoline alkaloids in C.chinensis var. brevisepala with those in other Coptis species. Meanwhile,molecular methods accurately identified 12 batches of C. chinensis var. brevisepala,9 batches of C. chinensis,4 batches of C. deltoidea,and 1 batch of C. teeta. Gradient elution was performed with Waters CORTECS C18 column( 4. 6 mm× 150 mm,2. 7 μm) and the mobile phase acetonitrile-water with 0. 4% formic acid. Mass spectrometry was conducted in ESI positive mode. The quantitative results showed that 8 main alkaloids had a good linear relationship within the concentration range( R2>0. 996),with the recovery rate of 95. 18%-105. 0% and the RSD of 0. 28%-3. 7%. Compared with that of other Coptis species,the rhizome of C. chinensis var. brevisepala had the highest contents of berberine and columbamine. The total content of the 8 alkaloids in C. chinensis var. brevisepala was similar to that in C. chinensis but higher than that of the other two species. PCA was performed to compare the alkaloids among the 4 species. Besides,the 8 alkaloids were evaluated in different parts of C. chinensis var. brevisepala. The results indicate that this method is reliable and efficient and can provide a reference for the quality research.

关键词(KeyWords): 宣黄连;液质联用技术;分子鉴定;定量分析;异喹啉生物碱
Coptis chinensis var.brevisepala;UPLC-QTOF-MS;molecular identification;quantitative analysis;isoquinoline alkaloids

Abstract:

Keywords:

基金项目(Foundation): 国家重点研发计划项目(2018YFC1706104)

作者(Authors): 王安琪;李西文;袁庆军;郭宁;杨滨;孙奕;
WANG An-qi;LI Xi-wen;YUAN Qing-jun;GUO Ning;YANG Bin;SUN Yi;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences;National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;

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