The Research Of Different Compatibility Of Mahuang Decoction Influence On Active Ingredients And Their Fingerprints Of Adjunctive Drug And Messenger Drug

Traditional Chinese medicine complex prescription which is the advantage and special feature of the traditional Chinese medicine choose suitable kinds of herbs and determine quantity of the herbs according to some kind of principle. Traditional Chinese medicine complex prescription was proved to be effective and safe through long using history.Traditional Chinese medicine complex prescription compatibility was first recorded in the "Huangdi's Internal Classics" and has formed a theoretic system. One of the representative theory is "QiQingHeHe" which acted as the general principle of the theoretic system.To probe into the study on traditional Chinese medicine complex prescription compatibility by the means of modern technology and methods is very important and very instructive for the modernization of traditional Chinese medicine and at the same time it can help to improve the development of the traditional Chinese medicine theories.There are two main methods that are widely used to study the compatibility of traditional Chinese medicine complex prescription. The first one is through discovering and comparing the pharmacology change between before and after using the medicine to evaluate the medicine's compatibility principal. The other one is through comparing the chemical ingredients changes between single medicine and complex prescription to discuss the mechanism of the medicine's compatibility.Ma Huang Decoction is composed of four kinds of popular Chinese herbs which are principal drug Herba ephedrae, ministerial drug ramulus cinnamomi, adjunctive drug Semen armeniacae amarum and messenger drug Radix et rhizoma glycyrrhizae. This complex prescription was come from the Treatise on Febrile Diseases written by famous doctor Zhang Zhong Jing in Chinese history. The decoction can induce sweat and dispelling exogenous evils and facilitate the flow of gastric qi to relieve asthma.We plan to discuss the relationship between compatibility changes and chemical ingredients, observe the quantity changes of amygdaloside between the single amygdala amara and Ma Huang decoction, the quantity changes of glycyrrhizic acid between the single Radix et rhizoma glycyrrhizae and Ma Huang decoction, and evaluate the changing rule of Radix et rhizoma glycyrrhizae's fingerprint.Objects: (1) To establish the measure method of amygdaloside which was the active ingredient in the amygdaloside adjunctive drug and evaluate the effect of the Ma Huang decoction compatibility on the quantity of the amygdaloside; (2) To establish the measure method of glycyrrhizic acid which was the active ingredient in the Radix et rhizorna glycyrrhizae messenger drug and evaluate the effect of the Ma Huang decoction compatibility on the quantity of the amygdaloside; (3) To establish the fingerprint of the Radix et rhizoma glycyrrhizae and the fingerprint of the Radix et rhizoma glycyrrhizae in the Ma Huang Decoction, and evaluate the effect of the Ma Huang decoction compatibility on the peak area size of the main same peaks of Radix et rhizoma glycyrrhizae fingerprint. four kinds of herbs respectively according to orthogonal design by traditional method and precipitate the decoction by means of ethanol.; (2)To measure the quantity of the amygdaloside in the sample of bitter Semen armeniacae amarum by means of HPLC-ELSD under the condition of chromatographic column Hypersil ODS, moving phase methanol: H2O(30:70), flow rate0.5ml/min, column temperature25℃, ELSD detector and the drift tube temperature is 98℃, the flow rate of carrier gas is 3.2 L·min^-1; (3)To measure the quantity of the amygdaloside in the samples by means of RP-HPLC under the condition of chromatographic column Hypersil ODS, moving phase methanol: H₂O(15:85), flow rate0.5ml/min, column temperature35℃, UV detector: 210nm; (4)To measure the quantity of the glycyrrhizic acid in the samples by means of RP-HPLC under the condition of chromatographic column Zorbax S_B-C₁₈, moving phase acetonitrile 1.5% gradient elution, flow rate0.8ml/min, column temperature is room temperature, DAD detector: 254nm; (5)To establish the HPLC fingerprints of Radix et rhizoma glycyrrhizae in different samples. Under the condition of chromatographic column Zorbax SB-C₁₈, gradient elution flow rate 0.6ml/min. room temperature, DAD detector: 254nm; (6)To analyze the measurement results by means of statistic methods.Results: (1)We establish the quantity measure method of amygdaloside in the bitter Semen armeniacae amarum by means of HPLC-ELSD. We find when the lamygdaloside's quantity is between 1.0 and 15.1ug there is a good linear correlation. The average recovery is 99.0% and the RSD is 2.9%; (2)We establish the quantity measure method of amygdaloside in Ma Huang decoction by means of HPLC. We find when the lamygdaloside's quantity is between 101 and 5030ng the there is a good linear correlation. The average recovery is 98.5% and the RSD is 1.17%. We also find that the other three kinds of herbs in the Ma Huang decoction has no influence on the level of amygdaloside; (3)We establish the quantity measure method of glycyrrhizic acid in Ma Huang decoction by means of HPLC. We find when the ammonium glycyrrhizinate level is between 101 and 5050ng there is a good linear correlation. The average recovery is 102.5% and the RSD is 1.81%. We also find that the other three kinds of herbs in the Ma Huang decoction have influence on the level of ammonium glycyrrhizinate; (4)We establish the Radix et rhizoma glycyrrhizae fingerprint and determine the 38 peaks by means of HPLC. This method has good stability and reproducibility that in accordance with the SSDA standards; (5)We establish the Radix et rhizoma glycyrrhizae fingerprint of Ma Huang decoction and determine eight peaks by means of HPLC. We find that there are seven of the eight peaks areas decrease significantly while six of the eight peaks areas increased significantly.Conclusions: We find the Ma Huang Decoction prescription compatibility can influence the level of amygdaloside and glycyrrhizic acid which is the active ingredients of bitter Semen armeniacae amarum and Radix et rhizoma glycyrrhizae respectively in a regular way. We find comparing with single herbs the level of chemical ingredients in amygdala amara and Radix et rhizoma glycyrrhizae has changed after compatibility. From our research we can make a conclusion that the changing chemical ingredients induced by compatibility lead to changing pharmacology effects and function of the Ma Huang decoction.New ideas: (1) Our research is the first one discussing Ma Huang Decoction prescription compatibility theory from the level of amygdaloside and glycyrrhizic acid in a way of dismantle the original prescription,discussing Ma Huang Decoction prescription compatibility theory influence on the Radix et rhizoma glycyrrhizae fingerprint; (2) Our research is the first one measuring the quantity of amygdaloside by means of HPLC-ELSD which provide a new method and can be used to detect the amygdaloside in both medical material and preparation containing bitter Semen armeniacae amarum.