Analysis of Norditerpenoid Alkaloids Extracted from Aconitum sinomantanum Nakai by Electrospray Ionization Tandem Mass Spectrometry

Electrospray ionization mass spectrometry（ESI-MS） was applied simultaneously in determining norditerpenoid alkaloids from the roots of Aconitum sinomantanum Nakai （RAS） based on molecular mass information. The tandem mass spectra（ ESI-MS^n） provided the alkaloidal structural information, through which the existence of these alkaloids was further confirmed. Accordingly, six known norditerpenoid alkaloids were simultaneously determined on the basis of their ESI-MS^n spectra. Furthermore, based on the diagnostic fragmentation pathways of alkaloidal MS^n, a rapid method for direct detection and characterization of alkaloids from an ethanolic extract of RAS was described.

Qualitative and Quantitative Analysis of Alkaloids in Cortex Phellodendri by HPLC-ESI-MS/MS and HPLC-DAD

A combined method of high performance liquid chromatograph-elecrtrospray-ionization mass spectrometer（HPLC-ESI-MS/MS） coupled with a photodiode array detector（HPLC-DAD） and principal component analysis（PCA） was applied to the qualitative and quantitative analyses of alkaloids in Cortex Phellodendri（CP） samples, and to the differentiation of two species of CP, Cortex Phellodendri Chinensis（CPC） and Cortex Phellodendri Amurensis（CPA）. Twenty-two peaks appeared in the HPLC-MS base peak chromatogram of CP detected by the HPLC-ESI-MS/MS analysis, and the alkaloids were identified according to the MSn data, the known MS fragmentation rules and the literature data. Five alkaloids including berberine, palmatine, jatrorrhizine, phellodendrine and magnoflorine were simultaneously determinated by the HPLC-DAD. Berberine was the primary component in all CP samples, and the contents of berberine and palmatine were exploited to be two critical parameters for effective discrimination between the two species of CP. The average content of berberine in CPC（58.75 mg/g） was higher than that in CPA（9.16 mg/g）, while the content of palmatine was less, only 0.25 mg/g in CPC and 4.19 mg/g in CPA. With the use of PCA, samples datasets were separated successfully into two different clusters corresponding to the two species, and berberine, pahnatine, phellodendrine and magnoflorine contribute most to the above mentioned calssifying . The proposed method oroved to be a useful tool in the aualitv control of Chinese herbal medicines.

ANALYSIS OF STRYCHNINE ALKALOIDS IN BIOLOGICAL SAMPLES BY THIN-LAYER CHROMATOGRAPHIC DENSITOMETRY AND DISTRIBUTION STUDY OF STRYCHNINE IN INTOXICATED RATS

A sensitive and rapid analytical procedure was established to detrmine strychnie alkaloids in diverse biological specimens. Samples were treated with hydrochloric acid to precipitate protein and scanned in thin-layer chromatograpic scanner in zig-zag dual-wavelength reflection mode.Detection sensitivity for strychniue was about 0. 05 μg. Quantification recoveries were between89. 2% and 99. 5% with RSD between 2. 92% and 13. 21 %. The method was applied to investigate thedistribution or strychnine in intoxicated rats. The results showed that there were remarkable difference in organs or tissues with the aigaest concentration 4. 56±1. 41μg/g in liver. Diversification experiment proves the method also could be used to analyze other alkaloids in biological samples if theconditions slightly modified.

Abnormal Secondary Growth and Histochemical Localization of Alkaloids in Root System of Aconitum flavum Hand.-Mazz.

[Objective] The purpose of this study was to clarify the structure,growth pattern and histochemical localization of alkaloids in root system of Aconitum flavum Hand.-Mazz.[Method] Paraffin sectioning and histochemistry were employed for performing the analysis in this study.[Result] The root system of Aconitum flavum Hand.-Mazz.consists of taproot,lateral root and adventitious root.The primary structure of root system is normal,but secondary structure shows abnormal.The cambium and the extra cambium of taproot form a ＂U＂-shaped secondary vascular bundle and tertiary bundle in abnormal secondary structure.The sieve tube group is made of little sieve tube group which is differentiated from primary phloem and cambium.Meanwhile,the secondary xylem in tuberous root also appears to be a ＂U＂ shape.Parenchyma cells of secondary phloem occupy most of the tuberous root.The sieve tube group of tuberous root is mainly differentiated from parenchyma cell of secondary phloem.[Conclusion] The difference in abnormal secondary structure of taproot and tuberous root are attributed to their varied cambium compose and activity pattern.Alkaloids are mainly accumulated in parenchyma cell of the inside cortex and between bundle in taproot,while parenchyma of secondary phloem and pith in tuberous root.

Diterpene Alkaloids of Aconitum kirinense Nakai I.Lepe-nine and Kirinine A

wo diterpene alkaloids, lepenine and kirinine A were isolated for the first time from the roots of Aconitum kirinense Nakai. Their structures have been established by IR, HRMS, NMR analysis and chemical reactions, and kininine A was found to be a new C 20 diterpene alkaloid.

Effect of processing on the alkaloids in Aconitum tubers by HPLC-TOF/MS

According to the Chinese Pharmacopoeia 2015, only processed Aconitum tubers can be clinically applied, and the effect of processing is unclear. This research aimed to explore the effect of processing on cardiac efficacy of alkaloids in Aconitum tubers. First, the chemical ingredients in unprocessed and processed Aconitum tubers were identified and compared by using high performance liquid chromatography time-of-flight mass spectrometry(HPLC-TOF/MS) and multivariate pattern recognition methods. Then the representative alkaloids in Aconitum tubers, aconitine, benzoylaconine, and aconine, which belong to diester-diterpenoid alkaloids,monoester-diterpenoid alkaloids, and amine-diterpenoid alkaloids, respectively, were selected for further validation of attenuated mechanism. Subsequent pharmacological experiments with aconitine, benzoylaconine,and aconine in SD rats were used to validate the effect of processing on cardiac functions. After processing the Aconitum tubers, it was found that the contents of diester-diterpenoid alkaloids were reduced, and those of monoester-diterpenoid alkaloids and amine-diterpenoid alkaloids were increased, suggesting that diesterditerpenoid alkaloids were transformed into monoester-diterpenoid alkaloids and amine-diterpenoid alkaloids.Through further decocting the aconitine in boiling water, it was confirmed that the three alkaloids could be progressively transformed. Pharmacological experiments with aconitine, benzoylaconine, and aconine in SD rats showed that aconitine at a dose of 0.01 mg/kg and aconine at a dose of 10 mg/kg enhanced the cardiac function, while benzoylaconine at a dose of 2 mg/kg weakened the cardiac function. The effect of processing is attributed to the transformation of the most toxic diester-diterpenoid alkaloids into less toxic monoesterditerpenoid alkaloids and amine-diterpenoid alkaloids.

Two New Norditerpenoid Alkaloids from Aconitum spicatum Stapf

Two new norditerpenoid alkaloids, spicatine A (1) and spicatine B (2) were isolated from the root of Aconitum spicatum. The new compounds were deduced on the basis of their spectral data (IR, HREIMS, EIMS, 1D, 2D-NMR). This is the first whole report on the isolation of diterpenoid alkaloids from the A.spicatum Stapf.

Two New Norditerpenoid Alkaloids of Aconitum nagarum

Two new norditerpenoid alkaloids were isolated from the roots of Aconitum nagarum.Their structures were elucidated as 13-hydroxyfranchetine 1 and 10-dehydroxyflavaconitine 2,respectively, by 1D and 2D NMR experiments.

Two new C18-norditerpenoid alkaloids from Aconitum delavayi

Further phytochemical investigation of the unique C 18-norditerpenoid alkaloids from the roots ofAconitum delavayi Franch led to the isolation of two new norditerpenoid alkaloids, delavaconitine F 1 and delavaconitine G 2. Their structures were determined from spectroscopic evidence.

Diterpenoid alkaloids from a Tibetan medicinal plant Aconitum richardsonianum var. pseudosessiliflorum and their cytotoxic activity

The chemical constituents from Aconitum richardsonianum var.pseudosessiliflorum were investigated.The roots of this plant were extracted three times with 90% EtOH at the room temperature.The ethanol extracts were combined and concentrated under reduced pressure to yield residue,which was suspended in water and successively partitioned with chloroform.The chloroform extraction was isolated and purified by silica gel and Sephadex LH-20 column chromatography.Six compounds were isolated and elucidated as delelatine（1）,isodelpheline（2）,3-acetylaconitine（3）,isoatisine（4）,nordhagenine A（5）and yunaconitine（6）.Compounds 1-5 were obtained from Aconitum Brunneum for the first time.Compound（1）showed significant cytotoxic activities（IC50=4.36 μM）against the human tumor cell line P388.

Two new C_(19)-diterpenoid alkaloids from roots Aconitum hemsleyanium var. atropurpureum

A new franchetine-type C19-diterpenoid alkaloid 3-hydroxyfranchetine 1 and a new aconitine-type C19-diterpenoid alkaloid atropurpursine 2 have been isolated from the roots of Aconitum hemsleyanium var. atropurpureum. The structures of these new alkaloids were established on the basis of spectral data.

Diterpenoid Alkaloids and One Lignan from the Roots of Aconitum pendulum Busch

Diterpenoid alkaloids have neroprotective activity.Herein,three napelline-type diterpenoid alkaloids 1-3,two aconitine-type diterpenoid alkaloids 4-5,and one isoquinline-type alkaloid 6,as well as one lignan glycoside 7,have been isolated from the roots of Aconitum pendulum Busch.Compounds 1 and 7 were new compounds,and their chemical structures were determined on the basis of nuclear magnetic resonance(NMR)spectra and mass spectrometry analysis.A ThT assay revealed that compound 2 showed significant disaggregation potency on the Aβ_(1−42)aggregates.

Investigation of Aconitine-type Alkaloids from Processed Tuber of Aconitum carmiechaeli by HPLC-ESI-MS/MS^n

Introduction Aconitine-type alkaloids isolated from the roots of Aconitum carmiechaeli show a potential toxicity and a broad spectrum of bioactivity[1-4].On the basis of the C8-substituent of C19-diterpenoid skeleton,aconitine-type alkaloids can be divided into diester-diterpenoid alkaloids（DDAs）,monoester-diterpenoid alkaloids（MDAs）,and lipo-alkaloids（Fig.1）.

Matrix Solid-phase Dispersion Extraction of Alkaloids from the Roots of Aconitum kusnezoffii Reichb

Matrix solid-phase dispersion（MSPD） was developed for the extraction of four alkaloids, including aconitine, mesaconitine, hypaconitine and deoxyaconitine, from the roots ofAconitum kusnezoffii Reichb. The determination of the analyte was carried out by high performance liquid chromatography with UV detection. The alkaline alumina was used as sorbent. The mixture of acetonitrile and water was used as elution solvent. Several extraction parameters, such as type of sorbent, the ratio of sample to solid support material, type of the elution solvent and the volume of the elution solvent were tested. Mean recoveries ranged from 93.16% to 102.73%, with relative standard deviations from 0.27% to 4.17%. With the extraction efficiency and time expenditure taken into account, MSPD extraction should be a comparatively good method.

Tangutidines A-C,Three Amphoteric Diterpene Alkaloids from Aconitum tanguticum

Three new diterpene alkaloids,tangutidines A-C(1-3),and four known alkaloids(4-7)were isolated from the whole plant of Aconitum tanguticum,from which amphoteric diterpene alkaloids(1-3)were obtained for the first time.The structures of 1-3 were elucidated by detailed interpretation of spectroscopic data,including MS and NMR data.All of them were evaluated for their cytotoxic activities.

New Alkaloids from Aconitum stapfianum

Nineteen alkaloids,including a new C19-diterpenoid alkaloid stapfianine A(1)and a new benzamide derivative stapfianine B(2)were isolated from the roots of Aconitum stapfianum.Their structures were established on the basis of extensive spectroscopic analyses(IR,HRESIMS,1D and 2D NMR).

Diterpenoid Alkaloids from the Aerial Parts of Aconitum flavum Hand.-Mazz

Sixteen diterpenoid alkaloids(DAs),including six aconitine-type alkaloids(5 and 9−13),seven 7,17-seco-aconitine-type alkaloids(1−4,6−8),two napelline-type alkaloids(14 and 15)as well as one veatchine-type alkaloid(16),were isolated from the aerial parts of Aconitum flavum Hand.-Mazz.In which,flavumolines A−D(1−4)were four new ones,and flavu-moline E(5)was reported as natural compound for the first time.Their chemical structures were elucidated by the analysis of extensive spectroscopic data.The inhibitory activities of these isolates on Cav3.1 low voltage-gated Ca^(2+)channel,NO production in LPS-activated RAW264.7cells,five human tumor cell lines,as well as acetylcholinesterase(AChE)were tested.

Profiling the Change of Key Chemical Ingredients in Combination of Aconitum carmichaeli Debx. and Bletilla striata (Thunb.) Reichb.f. by UPLC-QTOF/MS with Multivariate Statistical Analysis

In the present study, an ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS) based chemical profiling approach to rapidly evaluate chemical diversity after codecocting of the combination of Aconitum carmichaeli Debx.(wu-tou in Chinese, WT) and Bletilla striata(Thunb.) Reichb.f.(bai-ji in Chinese, BJ) incompatible pair. Two different kinds of decoctions, namely WT-BJ mixed decoction: mixed water extract of each individual herbs, and WT-BJ co-decoction: water extract of mixed two constituent herbs, were prepared. Batches of these two kinds of decoction samples were subjected to UPLC-QTOF/MS analysis, the datasets of tR-m/z pairs, ion intensities and sample codes were processed with supervised orthogonal partial least squared discriminant analysis(OPLS-DA) to holistically compare the difference between these two kinds of decoction samples. Once a clear classification trend was found in score plot, extended statistical analysis was performed to generate S-plot, in which the variables(tR-m/z pair) contributing most to the difference were clearly depicted as points at the two ends of "S", and the components that correlate to these ions were regarded as the most changed components during co-decocting of the incompatible pair. The identities of the changed components can be identified by comparing the retention times and mass spectra with those of reference compounds and/or tentatively assigned by matching empirical molecular formulae with those of the known compounds published in the literatures. Using the proposed approach, global chemical difference was found between mixed decoction and co-decoction, and hypaconitine, mesaconitine, deoxyaconitine, aconitine, 10-OH-mesaconitine, 10-OH-aconitine and deoxyhypaconitine were identified as the most changed toxic components of the combination of WT-BJ incompatible pair during co-decocting. It is suggested that this newly established approach could be used to practically reveal the possible toxic components changed/increased of the herbal combination taboos, e.g. the Eighteen Incompatible Medications(Shi Ba Fan), in traditional Chinese medicines.

Synergistic effect of pinellia total alkaloids and uncaria total alkaloids on anticonvulsant action in mice and rats

Aim To investigate the synergistic effect of the combination of pinellia total alkaloid （PTA） and uncaria total alkaloid （UTA）, and explore the mechanism of anticonvulsant action. Methods Anticonvulsant and toxic effect profiles of combinations of PTA with UTA, alone and at three fixed ratios of 1：4, 1 ：1, 4：1, were evaluated in maximal electroshock （MES）-induced seizures and acute toxicity test in mice. Respective ED50 and LD50 were calculated with Bliss＇s method. Their synergistic effect were evaluated by isobolographic analysis and allowed the determination of benefit indices （BI） for respective combinations. The model of convulsive rats kindled by penicillin topically injected into cortex was used to investigated the content of Glu, Asp, Gly and GABA in hippocampus using high performance liquid chromatography （HPLC）. Results Combinations of PTA and UTA at the ratio of 4：1 were synergistic in MES test and antagonistic in acute toxicity test, showing the best profile for combinations of PTA with UTA. In contrast, the ratios of 1 ：4 and 1 ： 1, despite synergistic in MES test, were additive in acute toxicity test. The 4：1 combination and two drugs alone significantly decreased Glu level and increased GABA level in the hippocampus, but the GABA level in the 4：1 combination group was higher than that in the two drugs alone groups. They did not have significant influence on the levels of ASp and Gly. Conclusion Combinations of PTA and UTA at 4：1 ratio demonstrated synergistic effect in anticonvulsant action and antagonistic effect in toxicity. The anticonvulsant mechanism might be related to decreasing the excitability of Glutamatergic neurons and increasing the inhibition of GABAergic neurons.