Hupehenine

Investigation of association of chemical profiles with the tracheobronchial relaxant activity of Chinese medicinal herb Beimu derived from various Fritillaria species.[Pubmed:28842340]

J Ethnopharmacol. 2018 Jan 10;210:39-46.

ETHNOPHARMACOLOGICAL RELEVANCE: Fritillariae Bulbus (Beimu in Chinese) is derived from the bulbus of many Fritillaria species (family Liliaceae), which has been used as an antitussive herb in traditional Chinese medicine for more than 2000 years. Due to the complexity of plant origins and significant variations in chemical profiles, the characterization of the profile of the major bioactive constituents and its association with pharmacological activity are important for the quality control of Beimu herbs from different origins. AIM OF THE STUDY: This study aims to investigate the distribution of major bioactive isosteroidal alkaloids in Beimu herbs of different origins and its correlation with the tracheobronchial relaxant activity. METHODS: Quantification of 7 main bioactive 5alpha-cevanine isosteroidal alkaloids, including ebeiedine, ebeiedinone, Hupehenine, isoverticine, verticine, verticinone and imperialine, in 23 Fritillaria species was performed using gas chromatography. The relaxant effect of different extracts of 4 commonly used Beimu herbs, namely Zhe-Beimu (F. thunbergii Miq.), Chuan-Beimu (F. cirrhosa D. Don), Hubei-Beimu (F. hupehensis Hsiao et K. C. Hsia) and Yi-Beimu (F. pallidiflora Schrenk), was evaluated using rat isolated tracheal and bronchial preparations pre-contracted with carbachol, the well established in vitro antitussive model. RESULTS: Amongst 23 Fritillaria species detected, significant variations of the types and quantities of 7 major isosteroidal alkaloids were determined, which served as an important indicator for the classification of different Beimu herbs with distinct geographic distributions. Based on the type and quantity of these alkaloids, different origins of Beimu could be clearly clustered into several subgroups by principal component analysis. Furthermore, both crude alkaloid and water extracts of all 4 Beimu herbs showed a dose-dependent tracheobronchial relaxation with different potencies. The total content of alkaloids (weight adjusted based on the activity of individual alkaloids) in Beimu extracts significantly correlated with their tracheobronchial relaxation effects (r(2) > 0.9, p < 0.001). CONCLUSIONS: The results demonstrated that the differences in chemical profile of major bioactive isosteroidal alkaloids and pharmacological activity of Beimu could be incorporated into a simple and unified method for quality control and potential prediction of activity of Beimu herbs from different origins.

[Comparative Analysis of Content of Four Alkaloids in Fritillaria hupehensis and Fritillaria ebeiensis var. purpurea].[Pubmed:27254925]

Zhong Yao Cai. 2015 Oct;38(10):2105-8.

OBJECTIVE: To establish an assay method for simultaneous determination of peimine, peiminine, peimissine and Hupehenine and to make a comparative analysis of the content of four alkaloids in Fritillaria hupehensis and Fritillaria ebeiensis var. purpurea for the first time. METHODS: A Unitary C18 column(250 mm x 4.6 mm, 5 mum) was chosen with acetonitrile-water (containing 0.05% diethylamine) as mobile phase in a gradient program. The column temperature was 35 degrees C and the flow-rate was 1.0 mL/min. RESULTS: There was high content of peiminine and the content of peimissine was inferior to peiminine in Fritillaria hupehensis. Relatively speaking, peimine and Hupehenine were much lower than the other two ingredients. Fritillaria ebeiensis var. purpurea also contained high levels of peiminine, the minimum content of peimine and equivalent content of peimissine comparing with Fritillaria hupehensis. In addition, it didn't contain Hupehenine in Fritillaria ebeiensis var. purpurea. CONCLUSION: This method is simple and fast, and it has good separation, reproducibility and reliable results. Also, it can be used as basis for the quality evaluation of Fritillaria hupehensis and Fritillaria ebeiensis var. purpurea.

Determination and validation of hupehenine in rat plasma by UPLC-MS/MS and its application to pharmacokinetic study.[Pubmed:26033449]

Biomed Chromatogr. 2015 Dec;29(12):1805-10.

In this work, a sensitive and selective ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for determination of Hupehenine in rat plasma was developed and validated. After addition of imperialine as an internal standard (IS), protein precipitation by acetonitrile-methanol (9:1, v/v) was used to prepare samples. Chromatographic separation was achieved on a UPLC BEH C18 column (2.1 x 100 mm, 1.7 microm) with 0.1% formic acid and acetonitrile as the mobile phase with gradient elution. An electrospray ionization source was applied and operated in positive ion mode; multiple reaction monitoring mode was used for quantification using target fragment ions m/z 416.3 --> 98.0 for Hupehenine, and m/z 430.3 --> 138.2 for IS. Calibration plots were linear throughout the range 2-2000 ng/mL for Hupehenine in rat plasma. Mean recoveries of Hupehenine in rat plasma ranged from 92.5 to 97.3%. Relative standard deviations of intra-day and inter-day precision were both <6%. The accuracy of the method was between 92.7 and 107.4%. The method was successfully applied to a pharmacokinetic study of Hupehenine after either oral or intravenous administration. For the first time, the bioavailability of Hupehenine was reported as 13.4%.

Development and validation of a liquid chromatography-mass spectrometry method for the determination of verticinone in rat plasma and its application to pharmacokinetic study.[Pubmed:20580328]

J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Aug 1;878(23):2067-71.

We have developed and validated a sensitive liquid chromatography-electrospray ionization-mass spectrometric (LC-ESI-MS) method for the quantification of verticinone, a major active constituent from Fritillaria hupehensis Hsiao et KC Hsia., in rat plasma. Verticinone and the internal standard (IS), Hupehenine, were extracted from plasma samples by a simple liquid-liquid extraction with ethyl acetate after being alkalified by 1M ammonia hydroxide. Chromatographic separation was achieved on a C(18) column using a gradient elution program with methanol and water as the mobile phase. The detection was performed by selected ion monitoring (SIM) mode via positive electrospray ionization (ESI) interface. The lower limit of quantification (LLOQ) was 0.1 ng/mL. The calibration curves were linear (r(2)>0.998) over the concentration range of 0.1-200 ng/mL. Within- and between-run precision was less than 6.5% and accuracy was within +/-10.7%. The validated method was applied to the pharmacokinetic study of verticinone in rats after a single oral administration of 1 mg/kg.

Analysis of hupehenine in the total alkaloids from Fritillaria hupehensis by HPLC-ELSD.[Pubmed:18278474]

J Huazhong Univ Sci Technolog Med Sci. 2008 Feb;28(1):118-20.

A reverse-phase high pressure liquid chromatography (HPLC) method with evaporative light scattering detection (ELSD) has been developed for the quantitative analysis of Hupehenine in the total alkaloids from Fritillaria hupehensis. Samples were analyzed on a reverse-phase column (Hypersil C-18) with a mobile phase of methanol:water:chloroform: triethylamine (85:15:1:0.6). The ELSD was set at the drift tube temperature of 68.3 degrees C and gas flow rate of 1.8 L/min. Hupehenine's retention time was 13.7 min with an asymmetry factor of 1.2. The validity of the method has been verified with linearity, limit of detection, accuracy and precision. The logarithmic linear curve was obtained from 8.936 to 134.04 microg/mL (r=0.9993). The detection limit (S/N>3) of Hupehenine was 1.79 microg/mL on the column. Intra-day RSD was 1.42% and inter-day RSD was 2.26% (3 days within a week). The average recovery of Hupehenine was 101.50%, and RSD was 1.62%.

[Studies on the chemical constituents from culbs of hybridized Bulbus Fritillariae Ussuriensis].[Pubmed:15706870]

Zhongguo Zhong Yao Za Zhi. 2004 Apr;29(4):331-4.

OBJECTIVE: To study the chemical constituents of hybridized Bulbus Fritillariae Ussuriensis. METHOD: The chemical constituents were isolated by silica column chromatography and their structures were identified by physical and chemical eveidences and spectral analysis (IR, 1H-NMR, 13C-NMR, 2D-NMR, MS). RESULT: Seven compounds were obtained and identified as (20S,25S)5alpha, 14alpha, 17beta-cevanine-6beta-hydroxy-3-one (hupehenirine, ZF1), (20S,25S)5alpha, 14alpha, 17beta-cevanine-3beta-hydroxy-6-one (hupehenizine, ZF2), (20R,25S)5alpha, 14alpha-cevanine-3beta,20beta-dihydroxy-6-one (peiminine, verticinone, ZF3), (20S,25S)5alpha, 14alpha, 17beta-cevanine-3beta, 6beta-dihydroxy (Hupehenine, ZF4), (20R,25S)5alpha, 14alpha-cevanine-3beta, 6beta, 20beta-trihydroxy (isoverticine, ZF5), (20R,25S)5alpha, 14alpha-cevanine-3beta, 6alpha, 20beta-trihydroxy (peimine, verticine, ZF6), (20S,25S)5alpha, 14alpha, 17beta-evanine-6beta-hydroxy-3beta-O-beta-D-glucoside (hupeheninoside, ZF7). CONCLUSION: Compounds ZF1-7 were isolated from hybridized Bulbus Fritillariae Ussuriensis for the first time.

Simultaneous determination of seven major isosteroidal alkaloids in bulbs of Fritillaria by gas chromatography.[Pubmed:10757299]

J Chromatogr A. 2000 Mar 24;873(2):221-8.

The present paper describes the development of a most simple, sensitive, and specific gas chromatographic method to date, for the direct determination of seven major bioactive isosteroidal alkaloids, namely ebeiedine, ebeiedinone, ebeienine, Hupehenine, isoverticine, verticine, verticinone and imperialine, in Fritillaria species, a commonly used antitussive traditional Chinese medicinal (TCM) herb. In the present study, a commercially available Supelco SAC-5 capillary column (30 m x 0.25 mm, 0.25 microm) specifically designed for the analysis of steroids was utilized for the direct determination of Fritillaria alkaloids. Calibration curves were obtained by spiking authentic compounds and the internal standard (solanidine) into herbal samples prior to extraction. Extraction was conducted simply by shaking the pre-alkalized diethyl ether solution (5.0 ml) containing dried herb (0.1 g) for 2 h. All calibration curves showed good linear regressions (r2>0.995) within test ranges. The assay was reproducible and accurate with the overall intra- and inter-day variation and accuracy of less than 10% and more than 90%, respectively. The developed GC method was successfully utilized to analyze seven major bioactive alkaloids in seven Fritillaria species, and the results demonstrate that this direct GC analytical method is suitable for the quality control of this commonly used antitussive TCM herb.