GC-MS Analysis of Petroleum Ether Fraction from the Ethanol Extract of Aconitum sinomontanum before and after Processing / 中国药房

OBJECTIVE：To compare the chemical constituents of petroleum ether fraction from ethanol extract of Aconitum sinomontanum before and after processing. METHODS ：After A. sinomontanum was purified with water ，the raw product decoction pieces were prepared ；the raw decoction pieces were steamed with licorice juice under high pressure to prepare processed decoction pieces of A. sinomontanum . The petroleum ether fractions of raw product and processed product were obtained after ultrasonic extraction with 95％ ethanol. The chemical constituents in the samples were analyzed by GC-MS. NIST 2014 mass spectrometry database was used to compare and match the components . The peak area normalization method was used to determine the relative percentage content of each component. RESULTS ：Before and after processing ，fatty acids and esters were the main components in the petroleum ether fraction from ethanol extract. Totally 18 chromatographic peaks were detected in the detection pieces of raw product，and 13 compounds were identified ，accounting for 94.60％ of the total content of volatile components. The components with relatively high content were （Z，Z，Z）-9，12，15-octadecatrienoic acid （26.13％），hexadecanoic acid ethyl ester （25.27％）， palmitoleic acid （10.84％），ethyl linoleic acid （10.67％），（Z，Z）-9，12-octadecenoic acid methyl ester （6.66％），pentadecanoic acid（5.11％）and so on. Totally 25 chromatographic peaks were detected in the decoction pieces of processed products，and 18 components were identified ，accounting for 82.40％ of the total content of volatile components. The components with relatively high content were palmitoleic acid （18.95％），（Z，Z）-9，12-octadecenoic acid methyl ester （17.93％），hexadecanoic acid ethyl ester（11.94％），（Z，Z，Z）-9，12，15-octadecatrienoic acid （10.54％），（Z，Z）-9，12-octadecenoic acid （5.51％），（Z）-11-hexadecanoic acid（5.30％）and so on. After processing ，7 new components were added ，5 of which were identified as （－）-eucalyptus globulus alcohol，ethyl 2-methyltetrade-canoate，6-methyl-4-phenylcoumarin，β-sitosterol，heptadecane. After processing ，no components disappeared，and the content of some components increased or decreased. CONCLUSIONS ：After processing ，the volatile components in the petroleum ether fraction from ethanol extract of A. sinomontanum are different ，and（－）-eucalyptus globulus alcohol and other components are added after processing.

Study of Acute Toxicity Different Extraction Parts of Aconitum sinomontanum Nakai on Mice / 中国中医药信息杂志

Objective To compare the toxicity of different extraction parts of Aconitum sinomontanum Nakai;To screen out"toxic sections"of Aconitum sinomontanum Nakai; To provide references for further study on toxicity components of Aconitum sinomontanum Nakai. Methods Systematic solvent method was used to extract the 95% ethanol extracts of Aconitum sinomontanum Nakai,and six different extraction fractions(petroleum ether,chloroform, ethyl acetate, butanol, alcohol and water) were obtained. Median lethal dose (LD50) and maximum dose method were used to conduct comparative study on acute toxicity of different extraction parts of Aconitum sinomontanum Nakai. Results Chloroform, water and butanol extractions in LD50of Aconitum sinomontanum Nakai were 89.65, 1805.40 and 24 409.41 mg/kg, and 95% confidence limits were 76.39~108.41, 1521.60~2240.00 and 20 422.54~24 246.95, respectively. The maximum dose of petroleum ether, ethyl acetate and alcohol extractions were 2686.01, 3108.13 and 28 376.21 mg/kg, respectively. Conclusion The maximum toxicity is the extracted section of chloroform, and the minimal toxicity is the extracted section of ethanol.

Spectrum and Toxicity Relationships between Multi-Wavelength HPLC Fingerprints and Acute Toxicity of Different Extracts of Aconitum sinomontanum Nakai / 中国药学杂志

OBJECTIVE To establish the HPLC multiple wavelength chromatographic fingerprints (MWCF) of different extracts of Aconitum sinomontanum Nakai (ASN) to clarify the attribution of the fingerprint peaks and their contribution to the acute toxicity. METHODS The experimental drugs (extracts of petroleum ether, chloroform, ethyl acetate, butanol, alcohol and water) were obtained by means of systematic solvent extraction from the 95% ethanol extract of ASN. The HPLC MWCF of different extracts of ASN were established by mean fingerprint method (MFM). The acute toxicity of different extracts in mice were carried out by measuring the median lethal dose (LD50) and maximum dose. The relationship between spectrum and toxicity was established by gray relational analysis. RESULTS The MWCF of different extracts of ASN were established. The acute toxicity of ASN was caused not only by lappacontine (LAP) and ranaconitine, the contribution of other diterpenoid alkaloids should not be neglected, and the contribution of different peaks to toxicity was ranked as (51, 38, 37, 35, 20) 3439323133. CONCLUSION The MWCF developed by MFM can maximally retain the fingerprint peaks, achieve fingerprint information maximization, and effectively improve fingerprint signal quality, thus providing a reference for the comprehensive quality evaluation of ASN. The relationship between the MWCF of different extracts and the acute toxicity is paralleled to some extent. And this will lay a foundation for the research of the toxicity mechanism of ASN.

One new diterpenoid alkaloid from roots of Aconitum sinomontanum / 中草药

Objective: To study the chemical constituents from the roots of Aconitum sinomontanum. Methods: Silica gel, Sephadex LH-20 column chromatography, high performance liquid chromatography, and other chromatographic techniques were used for separation and purification. The structures were elucidated by physiochemical methods and spectral data. Results: Five compounds were isolated from the 80% ethanol extract in the roots of A. sinomontanum, and their structures were identified as 6β,7β,8β,15α- tetrahydroxy-1α,14α,16β,18β-tetramethoxy-aconitan-19-en (1), delcosine (2), lepenine (3), napelline (4), and kirinine B (5). Conclusion: Compound 1 is a new compound named sinomontanum J. Compounds 2-5 are isolated from A. sinomontanum for the first time.

Herbal Textual Research on Aconitum Sinomontanum Nakai / 中国中医药信息杂志

Objective To provide herbalist basis for development of resource-related varieties through the study of herbal textual research of Aconitum sinomontanum Nakai, alias and source of production. Methods Description of Aconitum sinomontanum Nakai and its alias from the ancient herbal books was analyzed. Literature was searched to clarify modern research and conduct analysis. Results Ancient herbal textual records of Aconitum sinomontanum Nakai was not clear enough. The first records about Aconitum sinomontanum Nakai started from late Qing Dynasty, from which the source of production, medicinal parts, alias, harvest processing, efficacy and application of Aconitum sinomontanum Nakai were summed up. Conclusion Aconitum sinomontanum Nakai is the dry roots of Aconitum from Ranunculaceae, which has good medicinal prospects and development value, and can provide basis and guidance for the late clinical application and research.

HPLC Fingerprints and Quality Control of Processed Aconitum sinomontanum Nakai / 中国药学杂志

OBJECTIVE: To establish the fingerprint identification and assay for the quality analysis of processed Aconitum sinomontanum Nakai from different areas. METHODS: HPLC Gradient elution method was developed to establish the fingerprints for processed Aconitum sinomontanum Nakai, and the fingerprints were analyzed and compared by Chinese Materia Medica (CMM) Fingerprint Similarity Evaluation System (2012 edition), principal component analysis (PCA) and cluster analysis (CA). RESULTS: The common fingerprint for processed Aconitum sinomontanum Nakai fingerprints was established, and 18 common fingerprint peaks were identified. The similarity was greater than 0.90 among 10 batches of processed Aconitum sinomontanum Nakai medicinal herbs, and the contents of lappacontine and ranaconitine were determined. The samples from different areas could be classified into four groups, which reflected the quality characteristics of 10 batches of processed Aconitum sinomontanum Nakai from different areas. Four main components with cumulative contribution rate of 88.824% were selected by PCA, and seven chemical components were identified as the ones to determine the quality of processed Aconitum sinomontanum Nakai. CONCLUSION: This method, with good reproducibility and strong characteristics, can be used for the comprehensive quality evaluation of processed Aconitum sinomontanum Nakai.

Study on HPLC fingerprint and quality control of analgesic and anti-inflammatory effective parts in crude and processed roots of Aconitum sinomontanum / 中草药

Objective: To set up an analysis method of fingerprints for analgesic and anti-inflammatory effective parts from crude and processed roots of Aconitum sinomontanum (AS), and to discuss the chemical composition changes after processing that enhance the analgesic and anti-inflammatory effect. Methods: HPLC gradient elution method was developed to establish fingerprints for 10 batches of chloroform extract from crude and processed the roots of AS in different areas. And the fingerprint were analyzed and compared by Chinese Materia Medica (CMM) Fingerprint Similarity Evaluation System (2012 edition). Results: The fingerprints of chloroform extract from crude and processed the roots of AS were set up by HPLC. The gained 3 and 15 common peaks from crude and processed roots of AS, respectively. processed product added 12 peaks, including 1 peak, 2 peak, 7 peak increase were significant, accounting for the new peak area of 72.3%-84.5%. And determination of lappacontine and ranaconitine of chloroform extract from crude and proceed products, after processing the content were reduced, ranaconitine content reduced to the original one-third. Conclusion: This method with good reproducibility, and strong characteristic, and could be used for the full quality evaluation of analgesic and anti-inflammatory effect parts from crude and processed the roots of AS. To provide scientific basis for elucidating the chemical substance base and processing principle of crude and processed roots of AS.

Routine Examination of Aconitum sinomontanum Nakai and Content Determination of Lappacontine and Ranaconitine / 中国中医药信息杂志

Objective To study the routine examination of Aconitum sinomontanum Nakai and determine the contents of lappacontine and ranaconitine; To provide basis for establishing the quality standard of Aconitum sinomontanum Nakai.Methods Aconitum sinomontanum Nakai were collected from different areas.A method of TLC was used for qualitative discrimination. The methods in the Chinese Pharmacopoeia were adopted for the determination of moisture content, ash content and extractives. Determination of lappacontine and ranaconitine were performed by HPLC. Results The TLC showed that the spots were clear and the separation was good. Individual provisional standards:the moisture,total ash and acid-insoluble ash content of Aconitum sinomontanum Nakai were not more than 11.0%, 12.0%, and 7.0%, respectively; water soluble and alcohol soluble extractives were not less than 18.2% and 10.6%,respectively.The content of ranaconitine and lappacontine in Aconitum sinomontanum Nakai were not less than 0.125% and 0.815%, respectively. Conclusion The method established by the study is accurate and reliable,and can be used for quality evaluation of Aconitum sinomontanum Nakai.

Routine Examination of Aconitum sinomontanum Nakai and Content Determination of Lappacontine and Ranaconitine / 中国中医药信息杂志

Objective To study the routine examination of Aconitum sinomontanum Nakai and determine the contents of lappacontine and ranaconitine; To provide basis for establishing the quality standard of Aconitum sinomontanum Nakai.Methods Aconitum sinomontanum Nakai were collected from different areas.A method of TLC was used for qualitative discrimination. The methods in the Chinese Pharmacopoeia were adopted for the determination of moisture content, ash content and extractives. Determination of lappacontine and ranaconitine were performed by HPLC. Results The TLC showed that the spots were clear and the separation was good. Individual provisional standards:the moisture,total ash and acid-insoluble ash content of Aconitum sinomontanum Nakai were not more than 11.0%, 12.0%, and 7.0%, respectively; water soluble and alcohol soluble extractives were not less than 18.2% and 10.6%,respectively.The content of ranaconitine and lappacontine in Aconitum sinomontanum Nakai were not less than 0.125% and 0.815%, respectively. Conclusion The method established by the study is accurate and reliable,and can be used for quality evaluation of Aconitum sinomontanum Nakai.