附子配伍甘草、大黄、干姜调控药性物质基础研究
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摘要
目的：以中医药理论为指导,根据附子类中药主要以水煎剂入药的用药特点,选择了传统使用的生附子与现在使用的主流品种白附片、黑顺片为研究对象,系统研究附子与甘草、大黄、干姜以不同比例配伍(1：0.5、1：1、1：2)后其水煎液中指标成分含量的变化规律,并结合质谱的研究结果,从物质基础的角度揭示附子通过配伍减毒增效的作用机制,同时也为中药配伍调控药性提供一定的科学依据。
方法：采用紫外分光光度法(UV)、高效液相色谱法(HPLC),对附子中的指标成分(总生物碱、酯型生物碱、双酯型生物碱),甘草中的指标成分(甘草黄酮、甘草皂苷、甘草多糖),大黄中的指标成分(总蒽醌、结合型蒽醌、游离型蒽醌、大黄多糖)、干姜中的指标成分(辣椒素)进行含量测定；采用电喷雾质谱法(ESI)对乌头碱、新乌头碱、次乌头碱及其水解溶液和生附子配伍甘草、大黄、干姜的水煎液进行研究,通过质荷比(m/z)对其中的分子离子峰进行归属。
结果：1.附子配伍甘草：两者配伍后附子中的热性成分—附子总碱的含量略有升高,为2%左右；酯型生物碱和双酯型生物碱含量有显著的下降,最高降幅分别为63%和83%,而且升高和下降的幅度均与配伍甘草的量成正比。同时甘草中的黄酮和皂苷类成分含量也呈下降趋势,最高降幅分别为19%和55%,其下降的幅度也与配伍附子的量成正比。
2.附子配伍大黄：两者配伍后附子的附子总碱、酯型生物碱和双酯型生物碱的含量均有显著的下降,最高降幅分别为47%、64%和92%,而且下降的幅度均与配伍大黄的量成正比。同时大黄中的总蒽醌、结合型蒽醌、游离型蒽醌含量也呈下降趋势,最高降幅分别为64%、57%和75%,其下降的幅度也与配伍附子的量成正比。
3.附子配伍干姜：两者配伍后附子的附子总碱、酯型生物碱含量有一定程度的升高,最高升幅分别为31%和9%,而且升高的幅度与配伍干姜的量成正比；双酯型生物碱的含量与附子单煎液比基本无差别。同时干姜中的辣椒素含量也呈升高趋势,最高升幅分别为15%,其升高的幅度也与配伍附子的量成正比。
4.附子及其配伍药对的质谱研究：从乌头碱、新乌头碱和次乌头碱水解10分钟的质谱图上,观察到了C8位失去—分子乙酰基后的产物-苯甲酰乌头宁；从水解20分钟的质谱图上,观察到了C14位再失去—分子苯甲酰基后的产物-乌头原碱。从附子水煎液的质谱图上,观察到了乌头原碱、单酯型生物碱、双酯型生物碱和脂碱这四类生物碱成分.,共计15种。在配伍甘草和大黄的质谱图中,又观察到3种新的脂碱成分。
结论：本实验的研究结果从物质基础上证明了附子通过配伍能够改变药性,而且配伍的作用影响是相互的,其中配伍甘草是取其平性,缓和附子的大热、大毒；配伍大黄是取其寒性,除了能降低附子的毒性外,也能缓解大黄峻猛的泻下作用,变寒下为温下；配伍干姜是取其热性,共同增强回阳救逆的功效。综上所述,通过配伍调控药性是古人从临床实践中总结出来的,是中药独有的理论,对今天临床医生的用药具有非常重要的指导意义。
Objective:In the direction of Chinese Medicine theory, according to Radix Aconite Lateralis Prepapata mainly in the use of decoction solution, choose the traditional use of the Crude Radix aconiti lateralis preparata and now mainstream films White prepared lateral root of aconite, Black prepared lateral root of aconite as the research object, research Radix Aconite Lateralis Prepapata with Radix Glycyrrhizae, Radix et Rhizoma Rhei and Rhizoma Zingiberis compatibility different ratio (1:0.5,1:1,1:2) after the decoction of its ingredients target changes regular pattern, Combined with mass spectrometry results, From the chemical composition perspective reveals the Radix Aconite Lateralis Prepapata through the combined enhanced effect, reducing toxic mechanism. At the same time provide a scientific basis of Chinese herb control property regulation.
Methods:By ultraviolet spectrophotometry (UV), high-performance liquid chromatography (HPLC), determination of target ingredients of Radix Aconite Lateralis Prepapata(total alkaloids, monoeste-diter-penoidalkaloids, diester-diterpenoid alkaloids); Radix Glycyrrhizae(flavonoids,Saponins,Polysaccharide); Radix et Rhizoma Rhei(anthraquinone,anthraquinone-bound, Free anthraquinones, Polysaccharide), Rhizoma Zingiberis(capsaicin). Using electrospray ionization mass spectrometry (ESI) study the aconitine, mesaconitine, hypaconitine, and its hydrolysis solution, and Crude Radix aconiti lateralis preparata compatibility Radix Glycyrrhizae, Radix et Rhizoma Rhei and Rhizoma Zingiberis decoction solution. According to m/z confirm the molecular ion peak.
Results:1. Radix Aconite Lateralis Prepapata compatibility Radix Glycyrrhizae After water decoction, the content of total alkaloids increased slightly, about 2%; monoeste-diter-penoidalkaloids and diester-diterpenoid alkaloids content decreased obviously, the maximum drop of 63% and 83%, and directly proportional to compatibility the amount of Radix Glycyrrhizae. Flavonoids and saponins were decreased obviously, the maximum drop of 19% and 55%, and directly proportional to compatibility the amount of Radix Aconite Lateralis Prepapata.
2. Radix Aconite Lateralis Prepapata compatibility Radix et Rhizoma Rhei After water decoction, the content of total alkaloids, monoeste-diter-penoidalkaloids and diester-diterpenoid alkaloids decreased obviously, the maximum drop of 47%,64% and 92%, and directly proportional to compatibility the amount of Radix et Rhizoma Rhei. Anthraquinone, anthraquinone-bound and free anthraquinones were decreased obviously, the maximum decrease of 64%,57% and 75%, and directly proportional to compatibility the amount of Radix Aconite Lateralis Prepapata.
3. Radix Aconite Lateralis Prepapata compatibility Rhizoma Zingiberis After water decoction, the content of total alkaloids, monoeste-diter-penoidalkaloids increased, the maximum increase of 31% and 9%, and directly proportional to compatibility the amount of Rhizoma Zingiberis; diester-diterpenoid alkaloids content the same as the original. At the same time, capsaicin content increased,the maximum increase of 15%, and directly proportional to compatibility the amount of Radix Aconite Lateralis Prepapata.
4. Radix Aconite Lateralis Prepapata compatibility herb MS study From the aconitine, mesaconitine, hypaconitine hydrolysis of 10 minutes on the mass spectrum figure, we can see the product of they C_8_bit loss a acetyl,it's Benzoylaconine; From they hydrolysis of 20 minutes on the mass spectrum figure, we can see the product of they C_(14)_bit loss a benzoyl, it's aconine; From the Radix Aconite Lateralis Prepapata decoction solution have aconine, monoeste-diter-penoidalkaloids,diester-diterpenoid, lipid alkaloids, total 15,from the compatibility Radix Glycyrrhizae and Radix et Rhizoma Rhei, we observed that the three kinds of new lipid alkaloids.
Conclusion:The experiment results was based on material foundation evidence of Aconite can change herb property regulation through the compatibility herb, and mutual influence, compatibility Radix Glycyrrhiza use ping xing, ease Aconite big hot and toxicity; compatibility Radix et Rhizoma Rhei use han xing, except reducing Aconite toxicity; also ease Radix et Rhizoma Rhei strong role of diarrhea, variable hanxia to wenxia; compatibility Rhizoma Zingiberis use wen xing, together enhance the huiyangjiuli effection. In a word, compatibility change herb property regulation was eld come from the clinical practice, are unique theory of Chinese herb, of clinical doctors today have a very important guiding significance
方法：采用紫外分光光度法(UV)、高效液相色谱法(HPLC),对附子中的指标成分(总生物碱、酯型生物碱、双酯型生物碱),甘草中的指标成分(甘草黄酮、甘草皂苷、甘草多糖),大黄中的指标成分(总蒽醌、结合型蒽醌、游离型蒽醌、大黄多糖)、干姜中的指标成分(辣椒素)进行含量测定；采用电喷雾质谱法(ESI)对乌头碱、新乌头碱、次乌头碱及其水解溶液和生附子配伍甘草、大黄、干姜的水煎液进行研究,通过质荷比(m/z)对其中的分子离子峰进行归属。
结果：1.附子配伍甘草：两者配伍后附子中的热性成分—附子总碱的含量略有升高,为2%左右；酯型生物碱和双酯型生物碱含量有显著的下降,最高降幅分别为63%和83%,而且升高和下降的幅度均与配伍甘草的量成正比。同时甘草中的黄酮和皂苷类成分含量也呈下降趋势,最高降幅分别为19%和55%,其下降的幅度也与配伍附子的量成正比。
2.附子配伍大黄：两者配伍后附子的附子总碱、酯型生物碱和双酯型生物碱的含量均有显著的下降,最高降幅分别为47%、64%和92%,而且下降的幅度均与配伍大黄的量成正比。同时大黄中的总蒽醌、结合型蒽醌、游离型蒽醌含量也呈下降趋势,最高降幅分别为64%、57%和75%,其下降的幅度也与配伍附子的量成正比。
3.附子配伍干姜：两者配伍后附子的附子总碱、酯型生物碱含量有一定程度的升高,最高升幅分别为31%和9%,而且升高的幅度与配伍干姜的量成正比；双酯型生物碱的含量与附子单煎液比基本无差别。同时干姜中的辣椒素含量也呈升高趋势,最高升幅分别为15%,其升高的幅度也与配伍附子的量成正比。
4.附子及其配伍药对的质谱研究：从乌头碱、新乌头碱和次乌头碱水解10分钟的质谱图上,观察到了C8位失去—分子乙酰基后的产物-苯甲酰乌头宁；从水解20分钟的质谱图上,观察到了C14位再失去—分子苯甲酰基后的产物-乌头原碱。从附子水煎液的质谱图上,观察到了乌头原碱、单酯型生物碱、双酯型生物碱和脂碱这四类生物碱成分.,共计15种。在配伍甘草和大黄的质谱图中,又观察到3种新的脂碱成分。
结论：本实验的研究结果从物质基础上证明了附子通过配伍能够改变药性,而且配伍的作用影响是相互的,其中配伍甘草是取其平性,缓和附子的大热、大毒；配伍大黄是取其寒性,除了能降低附子的毒性外,也能缓解大黄峻猛的泻下作用,变寒下为温下；配伍干姜是取其热性,共同增强回阳救逆的功效。综上所述,通过配伍调控药性是古人从临床实践中总结出来的,是中药独有的理论,对今天临床医生的用药具有非常重要的指导意义。
Objective:In the direction of Chinese Medicine theory, according to Radix Aconite Lateralis Prepapata mainly in the use of decoction solution, choose the traditional use of the Crude Radix aconiti lateralis preparata and now mainstream films White prepared lateral root of aconite, Black prepared lateral root of aconite as the research object, research Radix Aconite Lateralis Prepapata with Radix Glycyrrhizae, Radix et Rhizoma Rhei and Rhizoma Zingiberis compatibility different ratio (1:0.5,1:1,1:2) after the decoction of its ingredients target changes regular pattern, Combined with mass spectrometry results, From the chemical composition perspective reveals the Radix Aconite Lateralis Prepapata through the combined enhanced effect, reducing toxic mechanism. At the same time provide a scientific basis of Chinese herb control property regulation.
Methods:By ultraviolet spectrophotometry (UV), high-performance liquid chromatography (HPLC), determination of target ingredients of Radix Aconite Lateralis Prepapata(total alkaloids, monoeste-diter-penoidalkaloids, diester-diterpenoid alkaloids); Radix Glycyrrhizae(flavonoids,Saponins,Polysaccharide); Radix et Rhizoma Rhei(anthraquinone,anthraquinone-bound, Free anthraquinones, Polysaccharide), Rhizoma Zingiberis(capsaicin). Using electrospray ionization mass spectrometry (ESI) study the aconitine, mesaconitine, hypaconitine, and its hydrolysis solution, and Crude Radix aconiti lateralis preparata compatibility Radix Glycyrrhizae, Radix et Rhizoma Rhei and Rhizoma Zingiberis decoction solution. According to m/z confirm the molecular ion peak.
Results:1. Radix Aconite Lateralis Prepapata compatibility Radix Glycyrrhizae After water decoction, the content of total alkaloids increased slightly, about 2%; monoeste-diter-penoidalkaloids and diester-diterpenoid alkaloids content decreased obviously, the maximum drop of 63% and 83%, and directly proportional to compatibility the amount of Radix Glycyrrhizae. Flavonoids and saponins were decreased obviously, the maximum drop of 19% and 55%, and directly proportional to compatibility the amount of Radix Aconite Lateralis Prepapata.
2. Radix Aconite Lateralis Prepapata compatibility Radix et Rhizoma Rhei After water decoction, the content of total alkaloids, monoeste-diter-penoidalkaloids and diester-diterpenoid alkaloids decreased obviously, the maximum drop of 47%,64% and 92%, and directly proportional to compatibility the amount of Radix et Rhizoma Rhei. Anthraquinone, anthraquinone-bound and free anthraquinones were decreased obviously, the maximum decrease of 64%,57% and 75%, and directly proportional to compatibility the amount of Radix Aconite Lateralis Prepapata.
3. Radix Aconite Lateralis Prepapata compatibility Rhizoma Zingiberis After water decoction, the content of total alkaloids, monoeste-diter-penoidalkaloids increased, the maximum increase of 31% and 9%, and directly proportional to compatibility the amount of Rhizoma Zingiberis; diester-diterpenoid alkaloids content the same as the original. At the same time, capsaicin content increased,the maximum increase of 15%, and directly proportional to compatibility the amount of Radix Aconite Lateralis Prepapata.
4. Radix Aconite Lateralis Prepapata compatibility herb MS study From the aconitine, mesaconitine, hypaconitine hydrolysis of 10 minutes on the mass spectrum figure, we can see the product of they C_8_bit loss a acetyl,it's Benzoylaconine; From they hydrolysis of 20 minutes on the mass spectrum figure, we can see the product of they C_(14)_bit loss a benzoyl, it's aconine; From the Radix Aconite Lateralis Prepapata decoction solution have aconine, monoeste-diter-penoidalkaloids,diester-diterpenoid, lipid alkaloids, total 15,from the compatibility Radix Glycyrrhizae and Radix et Rhizoma Rhei, we observed that the three kinds of new lipid alkaloids.
Conclusion:The experiment results was based on material foundation evidence of Aconite can change herb property regulation through the compatibility herb, and mutual influence, compatibility Radix Glycyrrhiza use ping xing, ease Aconite big hot and toxicity; compatibility Radix et Rhizoma Rhei use han xing, except reducing Aconite toxicity; also ease Radix et Rhizoma Rhei strong role of diarrhea, variable hanxia to wenxia; compatibility Rhizoma Zingiberis use wen xing, together enhance the huiyangjiuli effection. In a word, compatibility change herb property regulation was eld come from the clinical practice, are unique theory of Chinese herb, of clinical doctors today have a very important guiding significance
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