To investigate the chemical constituents of the roots of Aconitum taipaicum, silica gel column chromatography was used for the isolation and purification of compounds. A new norditerpenoid alkaloid, isodelelatine (1), along with five known alkaloids, atisine (2), delfissinol (3), liangshanine (4), hypaconitine (5) and delelatine (6) were isolated and identified. The structure of the new compound was elucidated on the basis of spectral data.
Aconitine ; analogs & derivatives ; chemistry ; isolation & purification ; Aconitum ; chemistry ; Alkaloids ; chemistry ; Diterpenes ; chemistry ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVETo study the processing principles of different processed products of Aconitum pendulum.

METHODUsing high performance liquid chromatography and acute toxicity test to compare the changes in chemical composition and toxicity of the roots and processed products of A. pendulum.

RESULTThe main toxic components of the roots of A. pendulum were aconitine, deoxyaconitine and 3-acetylaconitine. The contents of these three alkaloids were significantly reduced in processed products, while benzoylaconitine significantly increased. In addition, processed products emerged aconine, polyschistine-D, beyzoyldeoxyaconine, 16-epi-pyroaconitine and 16-epi-pyrodeoxyaconitine. From the structural analysis, these new emerged compounds transformed from the aconitine, deoxyaconitine and 3-acetylaconitine.

CONCLUSIONDifferent processing methods can reduce the toxicity of the roots of A. pendulum. Processing principle is ester hydrolysis and high-temperature pyrolysis.

Aconitine ; analogs & derivatives ; analysis ; Aconitum ; chemistry ; toxicity ; Animals ; Female ; Male ; Mice

OBJECTIVETo establish a capillary electrophoresis with field-amplified sample stacking method for the separation and determination of aconitine alkaloid in Guifudihuang pills.

METHODAn uncoated fused-silica capillary column (50 microm x 43 cm, effective length 35 cm) was used. The running buffer was 50 mmol x L(-1) phosphate electrolyte solution (pH 4.6)-methanol (8:2). The runing voltage was 10 kV and the capillary inlet was dipped in methanol for 5 s prior to electrokinetic injection (10 kV, 40 s). The detection wavelength was 235 nm.

RESULTThis method allowed 500 fold enrichment of aconitine alkaloid. A good linear relation was obtained in the range of 31.3-2 x 10(3) microg x L(-1) (r = 0.9996), with the detection limit of 9.4 microg x L(-1). The average recovery was 98.0% with the RSD of 2.6%.

CONCLUSIONThe method is simple, rapid and specific with high stacking efficiency; it provides a new reliable means for production and quality control of Guifudihuang pills.

Aconitine ; analysis ; Drugs, Chinese Herbal ; analysis ; chemistry ; Electrophoresis, Capillary ; methods ; Reproducibility of Results

OBJECTIVETo study the new hydrolysate of aconitine using HPLC-MS(n) and quantum chemistry calculation.

METHODThe HPLC method was applied in gradient elution program and the mass spectrometry was in positive ion mode. Geometries of the possible hydrolysates were optimized at DFT/6-31G(d) level.

RESULTThe new hydrolysate was found and its protonated molecularions was at m/z 482. The quantum chemistry calculation results show that the product of elimination reations at C8 and C15 got the lowest energy conformation. The compound at m/z 482 was decluced to be the hydrolysate of carbony at C15.

CONCLUSIONDelydration aconine was detected for the first time and the new hydrolysis pathways of aconitine in water were deduced.

Aconitine ; chemistry ; Chromatography, High Pressure Liquid ; Hydrolysis ; Mass Spectrometry ; Models, Molecular ; Molecular Conformation

This paper is aim to investigate the pharmacokinetics and absolute bioavailability of neoline in Beagle dogs, and provide a theoretical basis for further study. Ethyl acetate was used for liquid-liquid extracting after 10% ammonia alkalizing. The method of UPLC-Q-TOF-MS was established for the determination of neoline plasma concentrations. Beagle dogs were orally or intravenously administered with neoline for pharmacokinetic and absolute bioavailability study. Good linear relationship of neoline was found over the range of 0.1-4 mg x L(-1) (R2 = 0.9982) and 2-100 microg x L(-1) (R2 = 0.9945). Intra-and inter-day precision, expressed as the relativestandard (RSD) were less than 5.0%. Accuracy, expressed as the relative error (RE) was within 90.0%-115%. The recovery of neoline in dog plasma was more than 80%. After 6 mg x kg(-1) for ig and 1 mg x kg(-1) for iv administration of neoline, the main pharmacokinetic parameters were analyzed with Winnonlin software. t(1/2) were (313.88 +/- 63.18), (236.33 +/- 229.84) min, and AUC(0-infinity) were (58,027.40 +/- 14,132.69), (473,578.02 +/- 82,333.08) min x microg x L(-1) for ig and iv administration respectively. The absolute bioavail ability was (73.15 +/- 10.29) %. The method of UPLC-Q-TOF-MS described in the report was sensitive, reliable and specific, and suitable for pharmacokinetic study of neoline in Beagle dog. The high absolute bioavailability of neoline in dog suggested good absorption of neline which was worth of further investigation.
Aconitine ; analogs & derivatives ; chemistry ; pharmacokinetics ; Animals ; Biological Availability ; Dogs ; Drug Stability ; Female ; Male

OBJECTIVE: The transformations that occur in diterpenoid alkaloids during the process of sand frying for Chinese herbal medicine preparation have yet to be clarified. This study investigated the structural changes that take place in 3-acetylaconitine during a simulation of heat-processing and evaluated the toxicity and biological activity of the pyrolysis products. METHODS: The diterpenoid alkaloid 3-acetylaconitine was heated at 180 °C for 15 min to simulate the process of sand frying. The pyrolysis products were separated using column chromatography, and their structures were investigated using high-resolution electrospray ionization mass spectroscopy and nuclear magnetic resonance spectroscopy. Further, in vivo cardiotoxicity and acute toxicity of 3-acetylaconitine and its pyrolysis products were compared, and the aconitine-induced arrhythmia model was employed to evaluate the antiarrhythmic effect of the pyrolysis products. RESULTS: Two new diterpenoid alkaloids, pyroacetylaconitine and 16-epi-pyroacetylaconitine, a pair of epimers at C-16, were isolated. After comparing the structures of these compounds, possible transformation pathways were proposed. Compared with the prototype compound, 3-acetylaconitine, the cardiotoxicity and acute toxicity of the heat-transformed products were significantly decreased. In the biological activity assay, the two pyrolysis products exhibited an effective increase in ventricular premature beat latency, a reduction in the occurrence of ventricular tachycardia, as well as an increase in the rate of arrhythmia inhibition, implying strong antiarrhythmic activity. CONCLUSION Compared with 3-acetylaconitine, its pyrolysis products displayed lower toxicity and good antiarrhythmic effects; thus, they have potential for being developed into antiarrhythmic medicines. Please cite this article as: Wang YJ, Wang Y, Tao P. Structural characterization, in vivo toxicity and biological activity of two new pyro-type diterpenoid alkaloids derived from 3-acetylaconitine. J Integr Med. 2023; 21(3): 302-314.
Humans ; Aconitine/chemistry* ; Cardiotoxicity ; Sand ; Alkaloids/toxicity* ; Arrhythmias, Cardiac/drug therapy* ; Diterpenes/toxicity*

To study the variation of six ester-type alkaloids and characteristic fingerprints in the process from Radix Aconite Lateralis to Heishunpian and lay a foundation for the study of the processing principle of Heishunpian, HPLC. analysis was performed on a Phenomenex Gemini C18 (4.6 mm x 250 mm, 5 microm) with acetonitrile and 40 mmol x L(-1) ammonium acetate (adjusted to pH 10 with concentrated ammonia water) as mobile phase. The detection wavelength was set at 235 nm. The flow rate was set at 0.8 mL x min(-1) and the injection volume was 10-20 microL. Six ester-type alkaloids were determined and characteristic fingerprints of the process were established. As the process continues, the contents of diester diterpene alkaloids were decreased step by step, while the contents varia tion of monoester diterpene alkaloids were not obvious. Each sample showed significant difference in characteristic fingerprints. With the exception of 6 known monoester diterpene alkaloids and diester diterpene alkaloids, 13 peaks were marked in the characteristic fingerprints, of which the total change rule of the other 7 unknown peaks were similar with 3 diester diterpene alkaloids. The established method is accurate, reliable and repeatable, and can provide reference for revealing change rule of index components and illuminating processing principle in the process of Heishunpian.
Aconitine ; chemistry ; Aconitum ; chemistry ; Alkaloids ; chemistry ; Chemistry, Pharmaceutical ; methods ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Esters ; chemistry ; Molecular Structure

OBJECTIVETo find out hydrolysis regularity of aconitine.

METHODAconitine was air-tighted and hydrolyzed in water for 8, 12, 16, 20, 24 h. The hydrolysis products were analyzed by HPLC-MS. A major hydrolysis product was isolated by alumina oxide column chromatography, and identified by 1H and 13C-NMR spectra.

RESULTHPLC-MS analysis shows that the major hydrolysis products are benzoylaconine and aconine. The hydrolysis can mostly be completed within 20 hours.

CONCLUSIONDiester aconitum alkaloid can be changed to monoester aconitum and aconine alkaloids. Under the controled condition benzoylaconine is a major hydrolysis products.

Aconitine ; analogs & derivatives ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Hydrolysis ; Spectrometry, Mass, Electrospray Ionization ; Water ; chemistry

OBJECTIVETo study systematically the factors which affect separation and purification of the total alkaloids and mesaconitine with X -5 macroporous resin.

METHODWith the content of the total alkaloids and mesaconitine as parameters, the optimum condition of absorption and elution were studied in the process of the purification with X -5 macroporous resin.

RESULTThe X - 5 macroporous resin yielded the best separating efficiency when the concentration of the extracted solution was 1 g raw material per 1 mL, pH 12.0, the absorptive time of 6 hour and the volume of 95% ethanol (7BV pH 8) as the eluant; X -5 macroporous resins was used five times in a reproducible way. The rate of extraction and content of the total alkaloid were 80% and 30% respectively after purification with X - 5 macroprous resin.

CONCLUSIONThe method can increase the purity of mesaconitine and total alkaloids.

Aconitine ; analogs & derivatives ; analysis ; isolation & purification ; Aconitum ; chemistry ; Alkaloids ; analysis ; isolation & purification ; Ethanol ; chemistry ; Hydrogen-Ion Concentration ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Resins, Synthetic ; chemistry

OBJECTIVETo optimize the conditions of purifying the total alkaloids in Aconitum szechenyianum with macroporous adsorption resin, and compare the content of total alkaloids and aconitine in A. szechenyianum from different producing areas, in order to provide basis for further studies.

METHODThe orthogonal experiment method was adopted for optimizing the conditions of purifying the total alkaloids in A. szechenyianum with macroporous adsorption resin. The content of total alkaloids and aconitine were determined by using the titration method. The total alkaloids in A. szechenyianum from different producing areas were purified under optimum processing conditions. Aconitine was determined by HPLC.

RESULTDifferent processing conditions showed different influences on the purification of total alkaloids. The optimum conditions were resin type HPD-722, ethanol concentration of 80% , and ethanol elution volume of 80 mL x min(-1). The contents of aconitine in A. szechenyianum from different producing areas--Qinghai, Maxianshan, Ningxia and Yongdeng were 0.493 5, 0.883 5, 1.527 8, 1.664 4 mg x g(-1), respectively.

CONCLUSIONThe optimum processing conditions used in this essay could be used for purifying the total alkaloids and aconitine. A. szechenyianum from Yongdeng and Gansu contains the highest content of aconitine.

Aconitine ; chemistry ; isolation & purification ; Aconitum ; chemistry ; Adsorption ; Alkaloids ; chemistry ; isolation & purification ; China ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Porosity ; Resins, Synthetic ; chemistry