板蓝根和亚麻籽化学成分研究及活性评价
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摘要
复方板蓝根颗粒为《中华人民共和国卫生部药品标准》所收载的中药成方制剂,由板蓝根和大青叶两种中药材经水醇法提取后制备而成,具有清热解毒、凉血之功效,用于瘟病发热、出斑、风热感冒、咽喉肿烂、流行性乙型脑炎、肝炎和腮腺炎等;亚麻籽为亚麻科一年生草本植物,富含多种营养和保健成分,具有多种保健功能,作为原料已广泛应用于保健品和功能性食品的生产中。
本论文对复方板蓝根颗粒制成颗粒剂前的复方板蓝根提取物和亚麻籽的化学成分和活性进行了系统的研究,主要包括以下几个方面:
1.采用系统溶剂萃取的方法,将复方板蓝根提取物及其单味药逐级分离,得到了样品B1、B2、B3、B4、B、D1、D2、D3、D4、D、FF1、FF2、FF3、FF4和FF共15个化学组分,利用高效液相色谱与质谱联用技术、电喷雾多级串联质谱技术并结合高分辨质谱技术,对复方板蓝根提取物及其单味药各化学组分的化学成分进行了分析。
2.建立了复方板蓝根提取物各化学组分的质谱或色谱指纹图谱分析方法,并采用高效液相色谱技术和紫外分光光度法对复方板蓝根提取物各化学组分的主要化学成分进行了定量分析。
3.对复方板蓝根提取物的抗病毒活性、抗炎活性、抗菌活性、解热活性和免疫增强活性进行了评价,确定了复方板蓝根提取物中具有抗病毒、抗炎、抗菌、解热和免疫增强活性的化学组分。
4.对复方板蓝根提取物及其单味药的抗氧化活性进行了评价,确定了复方板蓝根提取物及其单味药具有抗氧化活性的化学组分。
5.对天然产物亚麻籽在发芽过程中的化学成分进行了定性和定量分析,并对亚麻籽发芽过程中的抗氧化活性进行了评价。
本论文的研究结果为复方板蓝根颗粒药效物质基础的研究、质量控制标准的制定以及谱效关系的研究奠定了基础。为亚麻籽作为功能性食品和保健品的原料提供了理论依据。
Chemical Composition and Bioactivity of Radix Isatidis and Flaxseed
The chemical components in traditional Chinese medicine (TCM) are responsible for their therapeutic effects and are very important in the modernization of TCM. High performance liquid chromatography (HPLC) and mass spectrometry (MS) techniques have been proved to be a powerful tool for the analysis of TCM due to the high efficiency, sensitivity, speed, resolution and low level of sample consumption. Especially with the development of high performance liquid chromatography coupled to mass spectrometry (HPLC-MS), it has a provided direct, rapid and effective method for the identification of known compounds, structural characterization of unknown compounds and screening of bioactive components in TCM. In this paper, we have studied on the chemical composition and bioactivity of extract of compound Indigowoad Root and flaxseed, including five parts.
1、Analyze chemical composition of extract of compound Indigowoad Root and its single herbal medicines.
We separated different chemical composition by systemic solvent step by step extraction from extract of compound Indigowoad Root and its single herbal medicines. Then we obtained the fifteen samples B1, B2, B3, B4, B, D1, D2, D3, D4, D, FF1, FF2, FF3, FF4 and FF. HPLC-MS has been applied successfully to the analyze the chemical composition from ethyl and n-BuOH extracts of extract of compound Indigowoad Root and its single herbal medicines. Twelve compounds were found in extracts from ethyl of extract of compound Indigowoad Root and its single herbal medicines, which are indoles, quinazolinones, organic acids and porphyrin; Eighteen compounds were found in extracts from n-BuOH of extract of compound Indigowoad Root and its single herbal medicines, which are nucleosides, purines, amino acids and flavonoids; Eleven compounds were found in extracts from 95% ethanol and water of extract of compound Indigowoad Root and its single herbal medicines, which are carbohydrates, amino acids, organic acids and two primary products of the maillard reaction.
2、The quantitative and fingerprint chromatograms analysis of chemical composition of extract of compound Indigowoad Root.
The MS and HPLC fingerprint chromatograms analytical methods had been developed and utilized to analyze the ethyl, n-BuOH, 95% ethanol and water extracts of extract of compound Indigowoad Root. There were ten common fingerprint peaks found in ethyl extracts of extract of compound Indigowoad Root, nineteen common fingerprint peaks found in n-BuOH extracts of extract of compound Indigowoad Root, nine common fingerprint peaks found in 95% ethanol extracts of extract of compound Indigowoad Root and seven common fingerprint peaks found in water extracts of extract of compound Indigowoad Root. Good similarities with correlation coefficients over 93% were found between the extracts from different sources of extract of compound Indigowoad Root. It showed that the extract of compound Indigowoad Root from different sources had similar chemical characteristics, and it also reflected the Indigowoad Root and Indigowoad Leaf from different sources had homologies and their chemical composition were similar after the medicine compatibility. The results of quantitative analysis showed that the contents of nucleosides, purines and flavonoids in extracts of extract of compound Indigowoad Root from different sources had little differences. The fingerprint chromatograms and quantitative analysis of cytidine, adenine. uridine, guanosine, adenosine, isovitexin and total flavonoids can be used for the quality evaluation of extract of compound Indigowoad Root, and the results were also useful for the studies on the spectrum-effect relationships of extract of compound Indigowoad Root.
3、Pharmacodynamic study of the extract of compound Indigowoad Root.
In this work, we have studied the anti-virus, anti-inflammatory, antibacterial, antipyretic and immunity enhancement activity of extract of compound Indigowoad Root. The results showed that the components of anti-virus are mainly existed in the middle polarity part. The components of anti-inflammatory are mainly existed in the big polarity part. The components of antibacterial are mainly existed in the small and middle polarity part. The ones of antipyretic and immunity enhancement are mainly existed in the big polarity part.
The results of chemical composition, fingerprints and pharmacodynamic of extract of compound Indigowoad Root showed that nucleosides, purines, amino acids and flavonoids might be the main anti-viral components. Carbohydrates, amino acids and organic acids might be the main anti-inflammatory components. Indoles, quinazolinones, organic acids, porphyrin, carbohydrates and amino acids might be the main anti-bacterial components. Carbohydrates, amino acids and organic acids might be the main active ingredient for antipyretic and immunity enhancement. The results were useful for the quality control of the extract of compound Indigowoad Root and the bioactive material basis of this prescription.
4、Antioxidant activity of extract of compound Indigowoad Root and its single herbal medicines.
The antioxidant activity of extract of compound Indigowoad Root and its single herbal medicines had been successfully investigated by the ferric reducing antioxidant power (FRAP) and radical scavenging (DPPH) methods. The results showed that the components of antioxidant activity are mainly existed in the small polarity part. Indoles, quinazolinones, organic acids and porphyrins might be the main antioxidative components.
5、Changes in phytochemicals and antioxidant activity of flaxseed during germination.
The chemical composition of flaxseed sprouts and its alkaline and acid hydrolysis products were analyzed by HPLC-MSn. The total flavonoid content (TFC) and total phenolic content (TPC) of flaxseed sprouts and its alkaline and acid hydrolysis products were measured by ultraviolet spectrophotometry methods. The antioxidant activities of flaxseed sprouts and its alkaline and acid hydrolysis products were determined by the FRAP, DPPH and oxygen radical absorbance capacity (ORAC). The mainly chemical components in flaxseed sprouts were sucrose, nucleosides and flavonoids, and the ones in alkaline and acid hydrolysis products were phenolics and lignans. The TFC and TPC values of flaxseed sprouts and its alkaline and acid hydrolysis products were increased from day 0 to day 8, and the antioxidant activity were positively correlated with the TFC and TPC values. Germination was shown to be a good process to increase the flavonoid content of flaxseeds as well as the antioxidant activity.
本论文对复方板蓝根颗粒制成颗粒剂前的复方板蓝根提取物和亚麻籽的化学成分和活性进行了系统的研究,主要包括以下几个方面:
1.采用系统溶剂萃取的方法,将复方板蓝根提取物及其单味药逐级分离,得到了样品B1、B2、B3、B4、B、D1、D2、D3、D4、D、FF1、FF2、FF3、FF4和FF共15个化学组分,利用高效液相色谱与质谱联用技术、电喷雾多级串联质谱技术并结合高分辨质谱技术,对复方板蓝根提取物及其单味药各化学组分的化学成分进行了分析。
2.建立了复方板蓝根提取物各化学组分的质谱或色谱指纹图谱分析方法,并采用高效液相色谱技术和紫外分光光度法对复方板蓝根提取物各化学组分的主要化学成分进行了定量分析。
3.对复方板蓝根提取物的抗病毒活性、抗炎活性、抗菌活性、解热活性和免疫增强活性进行了评价,确定了复方板蓝根提取物中具有抗病毒、抗炎、抗菌、解热和免疫增强活性的化学组分。
4.对复方板蓝根提取物及其单味药的抗氧化活性进行了评价,确定了复方板蓝根提取物及其单味药具有抗氧化活性的化学组分。
5.对天然产物亚麻籽在发芽过程中的化学成分进行了定性和定量分析,并对亚麻籽发芽过程中的抗氧化活性进行了评价。
本论文的研究结果为复方板蓝根颗粒药效物质基础的研究、质量控制标准的制定以及谱效关系的研究奠定了基础。为亚麻籽作为功能性食品和保健品的原料提供了理论依据。
Chemical Composition and Bioactivity of Radix Isatidis and Flaxseed
The chemical components in traditional Chinese medicine (TCM) are responsible for their therapeutic effects and are very important in the modernization of TCM. High performance liquid chromatography (HPLC) and mass spectrometry (MS) techniques have been proved to be a powerful tool for the analysis of TCM due to the high efficiency, sensitivity, speed, resolution and low level of sample consumption. Especially with the development of high performance liquid chromatography coupled to mass spectrometry (HPLC-MS), it has a provided direct, rapid and effective method for the identification of known compounds, structural characterization of unknown compounds and screening of bioactive components in TCM. In this paper, we have studied on the chemical composition and bioactivity of extract of compound Indigowoad Root and flaxseed, including five parts.
1、Analyze chemical composition of extract of compound Indigowoad Root and its single herbal medicines.
We separated different chemical composition by systemic solvent step by step extraction from extract of compound Indigowoad Root and its single herbal medicines. Then we obtained the fifteen samples B1, B2, B3, B4, B, D1, D2, D3, D4, D, FF1, FF2, FF3, FF4 and FF. HPLC-MS has been applied successfully to the analyze the chemical composition from ethyl and n-BuOH extracts of extract of compound Indigowoad Root and its single herbal medicines. Twelve compounds were found in extracts from ethyl of extract of compound Indigowoad Root and its single herbal medicines, which are indoles, quinazolinones, organic acids and porphyrin; Eighteen compounds were found in extracts from n-BuOH of extract of compound Indigowoad Root and its single herbal medicines, which are nucleosides, purines, amino acids and flavonoids; Eleven compounds were found in extracts from 95% ethanol and water of extract of compound Indigowoad Root and its single herbal medicines, which are carbohydrates, amino acids, organic acids and two primary products of the maillard reaction.
2、The quantitative and fingerprint chromatograms analysis of chemical composition of extract of compound Indigowoad Root.
The MS and HPLC fingerprint chromatograms analytical methods had been developed and utilized to analyze the ethyl, n-BuOH, 95% ethanol and water extracts of extract of compound Indigowoad Root. There were ten common fingerprint peaks found in ethyl extracts of extract of compound Indigowoad Root, nineteen common fingerprint peaks found in n-BuOH extracts of extract of compound Indigowoad Root, nine common fingerprint peaks found in 95% ethanol extracts of extract of compound Indigowoad Root and seven common fingerprint peaks found in water extracts of extract of compound Indigowoad Root. Good similarities with correlation coefficients over 93% were found between the extracts from different sources of extract of compound Indigowoad Root. It showed that the extract of compound Indigowoad Root from different sources had similar chemical characteristics, and it also reflected the Indigowoad Root and Indigowoad Leaf from different sources had homologies and their chemical composition were similar after the medicine compatibility. The results of quantitative analysis showed that the contents of nucleosides, purines and flavonoids in extracts of extract of compound Indigowoad Root from different sources had little differences. The fingerprint chromatograms and quantitative analysis of cytidine, adenine. uridine, guanosine, adenosine, isovitexin and total flavonoids can be used for the quality evaluation of extract of compound Indigowoad Root, and the results were also useful for the studies on the spectrum-effect relationships of extract of compound Indigowoad Root.
3、Pharmacodynamic study of the extract of compound Indigowoad Root.
In this work, we have studied the anti-virus, anti-inflammatory, antibacterial, antipyretic and immunity enhancement activity of extract of compound Indigowoad Root. The results showed that the components of anti-virus are mainly existed in the middle polarity part. The components of anti-inflammatory are mainly existed in the big polarity part. The components of antibacterial are mainly existed in the small and middle polarity part. The ones of antipyretic and immunity enhancement are mainly existed in the big polarity part.
The results of chemical composition, fingerprints and pharmacodynamic of extract of compound Indigowoad Root showed that nucleosides, purines, amino acids and flavonoids might be the main anti-viral components. Carbohydrates, amino acids and organic acids might be the main anti-inflammatory components. Indoles, quinazolinones, organic acids, porphyrin, carbohydrates and amino acids might be the main anti-bacterial components. Carbohydrates, amino acids and organic acids might be the main active ingredient for antipyretic and immunity enhancement. The results were useful for the quality control of the extract of compound Indigowoad Root and the bioactive material basis of this prescription.
4、Antioxidant activity of extract of compound Indigowoad Root and its single herbal medicines.
The antioxidant activity of extract of compound Indigowoad Root and its single herbal medicines had been successfully investigated by the ferric reducing antioxidant power (FRAP) and radical scavenging (DPPH) methods. The results showed that the components of antioxidant activity are mainly existed in the small polarity part. Indoles, quinazolinones, organic acids and porphyrins might be the main antioxidative components.
5、Changes in phytochemicals and antioxidant activity of flaxseed during germination.
The chemical composition of flaxseed sprouts and its alkaline and acid hydrolysis products were analyzed by HPLC-MSn. The total flavonoid content (TFC) and total phenolic content (TPC) of flaxseed sprouts and its alkaline and acid hydrolysis products were measured by ultraviolet spectrophotometry methods. The antioxidant activities of flaxseed sprouts and its alkaline and acid hydrolysis products were determined by the FRAP, DPPH and oxygen radical absorbance capacity (ORAC). The mainly chemical components in flaxseed sprouts were sucrose, nucleosides and flavonoids, and the ones in alkaline and acid hydrolysis products were phenolics and lignans. The TFC and TPC values of flaxseed sprouts and its alkaline and acid hydrolysis products were increased from day 0 to day 8, and the antioxidant activity were positively correlated with the TFC and TPC values. Germination was shown to be a good process to increase the flavonoid content of flaxseeds as well as the antioxidant activity.
引文
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