Studies On The Chemical Constituents And Quality Control Of Lateral Roots Of Aconitum Carmichaelii Debx

Lateral Roots of Aconitum carmichaelii Debx.(Fu zi) are the processed products of the Aconitum carmichaelii Debx., according to different processing methods can be divided into salted aconite root, black prepared lateral root of aconite and sliced white aconite, etc, which are widely distributed and cultivated in southwestern China, especially Sichuan Province. Lateral Roots of A. carmichaelii are often used as cardiac agents for keeping the heart pumping and rescuing the heart deficiency.In order to clarify the medicinal material base of this herb and the safety of clinical medication, the chemical constituents, toxic ingredients and quality control of Lateral Roots of A. carmichaelii were studied systematically in this paper. The results could be seen as follows:1. Research of the chemical compositions in Lateral Roots of A. carmichaelii.Firstly, the EtOH extraction of the Lateral roots of A. carmichaelii were isolated and purified with a variety of chromatographic separation technology, and twenty-five chemical constituents were obtained. And twenty-two chemical constituents were identified by analyzing the physicochemical properties and the spectral datas of these obtained constituents. Those compounds were identified as follows:aconitamide (1), sesamin (2), episesamin (3), isolariciresinol (4), honokiol (5), pinoresinol (6), β-Fruf(2-6)-α-Glu (7), p-hydroxy-cinnamic acid (8), salicylic acid (9), linoleic acid (10), fuziline (11), neoline (12), isotalatizidine (13), karakoline (14), songorine (15),8-OEt-14-benzoylmesaconine (16), mesaconitine (17), hypaconitine (18), aconitine (19), benzoylmesaconine (20), benzoylhypaconitine (21), benzoylaconitine (22). Among them, a new chemical compound (1), and eight chemical compounds (2-9) were isolated from Lateral Roots of A. carmichaelii for the first time. 2. The study on vitro toxicity of different alkaloids isolated from Lateral Roots of A. carmichaelii.Pharmacological experiments were made by comparing the damages of neonatal rat cardiac myocytes caused by diester-diterpenoid alkaloids (aconitine^benzoylmesaconine and benzoylaconitine) and monoester-diterpenoid alkaloid (benzoylmesaconitine) respectively, and the results showed that the poisonousness of the three diester-diterpenoid alkaloids decreased in the order of mesaconitine> aconitine> hypaconitine. However, the poisonousness of the above diester-diterpenoid alkaloids was stronger than that of the monoester-diterpenoid alkaloid benzoylmesaconitine.3. Composition analysis of the alkaloids in Lateral Roots of A carmichaelii.Online composition analysis of the alkaloids in the Lateral Roots of A. carmichaelii was founded by UPLC/Q-TOF-MSE method, and fifteen alkaloids were identified by elementary composition, mass spectrometry fragments and revelent literatures. The results laid the foundation for the research of the chemical compositions and the quality control of the herbal materials.4. UPLC-MS method for simultaneous measurement of seven alkaloid ingredients with single marker in Lateral Roots of A. carmichaelii.A new quantitative method was established for simultaneous determination of multi-component (songorine, fuziline, neoline, benzoylmesaconitine,14-benzoylaconitine, mesaconitine, hypaconitine) in Lateral Roots of A. carmichaelii by using one chemical reference substance and validated its feasibilities. The quantitative analysis of multi-components by single-marker (QAMS) was established and validated with Lateral Roots of A. carmichaelii. Seven main ingredients were evaluated and their relative correlation factors (RCFs) were determined by UPLC-MS. The contents of14-benzoylaconitine in the samples of Lateral Roots of A. carmichaelii were calculated by using the external standard method, and the contents of the six other ingredients were calculated by their RCFs. The contents of these six ingredients in all samples were also determined by using the external standard method. For these six compounds, there was no significant difference between the quantitative results of the two methods. The method is simple, accurate and suitable to evaluate the contents of seven toxic alkaloids in Lateral Roots of A. carmichaelii.5. The research progress on chemical constituents, pharmacological activities and content determination methods of Lateral Roots of A. carmichaelii were reviewed, providing useful references for the further research.