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OBJECTIVE: To study the effects of three aconitine transporters in Caco-2 cells and tannin (tannic acid) in Terminalia on the transport of three di-ester aconitine (aconitine, meoaconitine and hypaconitine) in Aconitum chinensis. METHODS: The components were detected by UPLC/Q exactive MS in terms of the cumulative transshipment volume of three aconitine and the apparent permeability coefficient Papp as indicators to investigate the two-way transport behaviors of three aconitine in the Caco-2 cell model, as well as the proportion of tannic acid, and the changes of the transport behaviors of three aconitine. RESULTS: There was a positive correlation between the cumulative transshipment volume of aconitine and the incubation time. There was no statistical difference in Papp values of the three aconitine, and the efflux effect was significantly stronger than the absorption, with an external ratio close to 1.5.When the compatibility ratio of three aconitine with tannic acid was 1∶1 and 1∶0.5, the absorption of aconitine was significantly inhibited (P was 0.05), but the transport behavior with effluent was not significantly affected. CONCLUSION: Aconitine, meoaconitine, and hypaconitine in P. aeruginosa are mainly passive transporters, and may involve in efferent proteins, which are a kind of drug with good absorption.When mixed with tannic acid, the absorption and transshipment of three kinds of alkali were significantly reduced, which proves that Terminalia chebula could detoxify aconitum by inhibiting its absorption.
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Objective: To investigate the content changes of alkaloids in stems and leaves of Aconiti Radix at different growth period. Methods: The Phenomenex C18 column (150 mm × 4.6 mm, 5 μm) was used and 0.1% formic acid aqueous solution-acetonitrile was selected as mobile phase; The mass spectrum was scanned by ESI+ multiple reaction monitoring (MRM) mode. The HPLC-MS/MS method was established for the simultaneous determination of aconitine, mesaconitine, hypaconitine, indaconitine, benzoylaconine, benzoylmesaconine, benzoylhypaconitine, aconine, fuziline, neoline, talatisamine, songorine, higenamine, and salsoline in the stems and leaves of Aconite Radix. Combined with principal component analysis (PCA), the transfer rule of various alkaoids was tracked in the growth cycle of Aconiti Radix. Results: Methodological validation results showed that the linear range of the 14 compounds was good (r2 > 0.990 0). The limit of quantification was 2.27-18.27 ng/mL, and the average recovery was 94.73%-104.50%. The results showed that there was considerable amounts of alkaloids in stems and leaves of Aconiti Radix, and the total contents of alkaloids in stems were higher than those in leaves. In May, June, July, and August, the total content of alkaloids in stems was 0.087 1%, 0.182 8%, 0.141 0%, and 0.199 4% respectively, which showed a wave-like upward trend. The total content of alkaloids in leaves was 0.074 7%, 0.075 9%, 0.081 4%, and 0.058 9% respectively, which showed a trend of rising first and then decreasing, and the total content of alkaloids in leaves was highest in July. Conclusion: PCA found that the alkaloids content in stems and leaves showed different variation trend in the different period. The considerable alkaloids in stems reached the peak value at the harvest time. The amount of alkaloids is considerable, which has the potential and development value of becoming new medicinal resources.
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This present study is to develop an HPLC method for simultaneous determination of four diester diterpenoid alkaloids, beiwutine, mesaconitine, hypaconitine and aconitine in the leaves of Aconitum kusnezoffii, so as to provide evidence of the quality control of this herb. The four constituents were measured on a Waters XBridge CC₁₈ column(4.6 mmχ250 mm, 5 μm). The mobile phase was acetonitrile-40 mmol·L⁻¹ ammonium acetate solution(adjusted pH to 10.5 with ammonia solution)(33:67) with isocratic elution at a flow rate of 1.0 mL·min⁻¹; the detection wavelength was 235 nm; the column temperature was 30 °C, and the injection volume was 10 μL. Next, this contents of the four diester diterpenoid alkaloids in 12 samples were 0.025 5-0.088 5, 0.039 1-0.071 5, 0.026 6-0.081 0 and 0.008 12-0.031 2 mg·g⁻¹, respectively. Next, this method has been successfully applied to the analysis of A. kusnezoffii folium in different harvest periods. The contents of the four alkaloids decreased primarily, and then increased with the postponing of harvest. The established method is proved to be accurate and sensitive for the determination of alkaloids in A. kusnezoffii folium, and may be useful for the quality improvement of this herbal medicine. Moreover, these results indicated the scientific significance for the toxicity and the suitable harvest time of this herb.
Subject(s)
Aconitine , Aconitum , Chemistry , Chromatography, High Pressure Liquid , Diterpene Alkaloids , Drugs, Chinese Herbal , Chemistry , Phytochemicals , Plant Leaves , Chemistry , Plants, Medicinal , ChemistryABSTRACT
AIM To explore the anti-heart failure effects of compatibility of glycyrrhetinic acid,liquiritin and hypaconitine on apoptotic pathway in rats with chronic heart failure and the possible mechanism of action.METHODS Sixty rats were operated through transverse aortic constriction to create the chronic heart failure models.After four weeks,these rats were randomly divided into seven groups:sham group (n =6),model group (n =7),digoxin group (n =8),hypaconitine group (n =9),glycyrrhetinic acid + hypaconitine group (n =9),liquiritin + hypaconitine group (n =9),glycyrrhetinic acid + liquiritin + hypaconitine group (n =9).Then the rats were given medicines or saline perfusion by oral gavage for one week.On the 7th day,the rats were executed.Before the rats were killed,the blood samples were obtained and echocardiographic were carried on to get the ejection fraction (EF) and fractional shortening (FS) data;and the ELISA test was done for type B natriuretic peptide (BNP) changes;and myocardial histopathological examinations were performed on Bcl-2,Bax and Caspase-3.And the protein expressions of Fas and Fas-L were detected by Western blot.RESULTS Compared with the model group,the expression of Bcl-2 of all the compatibility groups except the hypaconitine group was higher than that of the model group;the expressions of Bax,Caspase-3,Fas and Fas-L,besides with the measurements of EF,FS and BNP were lower than those of the model group,and the glycyrrhetinic acid + liquiritin + hypaconitine group showed the best effect.CONCLUSION The compatibility of hypaconitine,glycyrrhetinic acid and liquiritin could down-regulate the levels of EF,FS and BNP,together with the protein expressions of pro-apoptosis,meanwhile,up-regulate the protein expressions of anti-apoptosis.This might be one of the mechanisms for the anti-apoptosis effects on chronic heart failure by compatibility of Glycyrrhizea radix et rhizoma (Gancao) and Acontti lateralis Radix praparata (Fuzi).
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Epithelial-mesenchymal transition (EMT) has been implicated in tumor invasion and metastasis and provides novel strategies for cancer therapy. Hypaconitine (HpA), a diester-diterpenoid alkaloid isolated from the root of the Aconitum species, exhibits anti-inflammatory, analgesic, and especially, cardiotoxic activities. Here, we reported the anti-metastatic potentials of HpA in transforming growth factor-β1 (TGF-β1)-induced EMT in lung cancer A549 cells. The cytotoxic effect of HpA was determined by MTT assay. A549 cells were treated with TGF-β1 with or without HpA co-treatment, and the morphological alterations were observed with a microscopy. The expression of E-cadherin, N-cadherin, and NF-κB was determined by both Western blotting and immunofluorescence analyses. The adhesion, migration, and invasion were detected with Matrigel, wound-healing, and transwell assays, respectively. The expression of Snail was determined by Western blotting. The expression of NF-κB p65, IκBα, and p-IκBα in nuclear and cytosolic extracts was assessed by Western blotting. The results showed that low concentration of HpA (<16 μmol·L) had no obvious cytotoxicity to A549 cells. Morphologically, TGF-β1 treatment induced spindle-shaped alteration in the cells. The upregulation of N-cadherin, NF-κB, and Snail and the downregulation of E-cadherin were detected after TGF-β1 treatment. The adhesion, migration and invasion abilities were also increased by TGF-β1. Besides, TGF-β1 induced expression of Snail in a time-dependent manner. Furthermore, TGF-β1 induced nuclear translocation of NF-κB p65. All these alterations were dramatically inhibited by HpA co-treatment. In addition, the NF-κB inhibitor PDTC showed similar inhibitory effect. In conclusion, these results showed that HpA inhibited TGF-β1-induced EMT in A549 cells, which was possibly mediated by the inactivation of the NF-κB signaling pathway, providing an evidence for anti-cancer effect of HpA.
Subject(s)
Humans , A549 Cells , Aconitine , Pharmacology , Active Transport, Cell Nucleus , Antineoplastic Agents, Phytogenic , Pharmacology , Cadherins , Cell Adhesion , Cell Movement , Dose-Response Relationship, Drug , Epithelial-Mesenchymal Transition , NF-kappa B , Metabolism , Neoplasm Invasiveness , Transforming Growth Factor beta1 , PhysiologyABSTRACT
Objective:To develop an HPLC gradient elution method for the simultaneous determination of fangchinoline,tetrandrine,mesaconitine,aconitine and hypaconitine in Huoxue Zhentong plaster.Methods::A Dikma-C18 (200 mm×4.6 mm,5 μm) chromatographic column was adopted,the mobile phase was methanol-acetonitrile (3∶1)(A)-0.06% diaethylamin solution (B) with gradient elution at a flow rate of 1.0 ml·min-1,the detection wavelength was 280 nm for fangchinoline and tetrandrine,and 235 nm for mesaconitine,aconitine and hypaconitine.The column temperature was set at 30 ℃,and the injection volume was 10 μl.Results::There was a good linear relationship when the content of fangchinoline,tetrandrine,mesaconitine,aconitine and hypaconitine was within the range of 7.490-149.800 μg·ml-1(r=0.999 9),14.610-292.200 μg·ml-1(r=0.999 8),4.150-83.000 μg·ml-1(r=0.999 2),5.250-105.000 μg·ml-1(r=0.999 6) and 5.140-102.800 μg·ml-1(r=0.999 9),respectively.The average recovery and the corresponding RSD were 99.87%(0.49%),97.79%(1.11%),96.97%(1.75%),98.60%(1.50%) and 97.94%(0.98%)(n=6),respectively.Conclusion:An HPLC gradient elution method is successfully established for the simultaneous determination of 5 components in Huoxue Zhentong plaster.The established method is simple,accurate and reliable,which is helpful to the quality control of Huoxue Zhentong plaster.
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AIM To explore the anti-heart failure effects of compatibility of glycyrrhetinic acid,liquiritin and hypaconitine on apoptotic pathway in rats with chronic heart failure and the possible mechanism of action.METHODS Sixty rats were operated through transverse aortic constriction to create the chronic heart failure models.After four weeks,these rats were randomly divided into seven groups:sham group (n =6),model group (n =7),digoxin group (n =8),hypaconitine group (n =9),glycyrrhetinic acid + hypaconitine group (n =9),liquiritin + hypaconitine group (n =9),glycyrrhetinic acid + liquiritin + hypaconitine group (n =9).Then the rats were given medicines or saline perfusion by oral gavage for one week.On the 7th day,the rats were executed.Before the rats were killed,the blood samples were obtained and echocardiographic were carried on to get the ejection fraction (EF) and fractional shortening (FS) data;and the ELISA test was done for type B natriuretic peptide (BNP) changes;and myocardial histopathological examinations were performed on Bcl-2,Bax and Caspase-3.And the protein expressions of Fas and Fas-L were detected by Western blot.RESULTS Compared with the model group,the expression of Bcl-2 of all the compatibility groups except the hypaconitine group was higher than that of the model group;the expressions of Bax,Caspase-3,Fas and Fas-L,besides with the measurements of EF,FS and BNP were lower than those of the model group,and the glycyrrhetinic acid + liquiritin + hypaconitine group showed the best effect.CONCLUSION The compatibility of hypaconitine,glycyrrhetinic acid and liquiritin could down-regulate the levels of EF,FS and BNP,together with the protein expressions of pro-apoptosis,meanwhile,up-regulate the protein expressions of anti-apoptosis.This might be one of the mechanisms for the anti-apoptosis effects on chronic heart failure by compatibility of Glycyrrhizea radix et rhizoma (Gancao) and Acontti lateralis Radix praparata (Fuzi).
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Objective: To study the fragmentation pathways of six aconitine-type alkaloids (aconitine, hypaconitine, mesaconitine, benzoylaconitine, benzoylhypaconitine, and benzoylmesaconitine) in mass spectra (MS). Methods: The samples were analyzed by liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). Results: In positive mode, The typical fragmentation pathways of the six aconitine-type alkaloids were mainly continuous loss of CH3OH and H2O. The characteristic fragmentation pathway of diester-diterpene type aconitine was losing an acetic acid molecule from C-8 position; However this fragmentation could not be observed in MS of mono-ester aconitine alkaloids. Conclusion: The study on fragmentation pathways could be adopted for the structural identification of the six aconitine-type alkaloids.
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The species in Aconitum L. are commonly used in China as well known toxic herbs. Its main components, such as aconitine, mesaconitine, and hypaconitine, have significant pharmacological activity, and are also its toxic components. As a result, the therapeutic dose and toxic dose are very close, and the clinical therapeutic window is narrow. Adverse reactions and poisoning incidents occur frequently in clinic, which limits its wide applications. Modern toxicology studies on the plants of Aconitum L. to make the toxicity and its clear mechanism have important significance for more reasonable clinical guidance and safety evaluation. This paper reviews the toxicity of the plants of Aconitum L. and its mechanism and provides a scientific basis for clinical use.
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<p>In 2013, we prescribed daiuzusen for 3 patients with intractable pain; pain from complex regional pain syndrome, colic pain of unknown origin after an abdominal operation, and colic pain from advanced colon cancer and ileus. A dose of daiuzusen (containing uzu 0.5-2 g) quickly relieved their pain in several minutes. Another common symptom was “cold” in their bowel or extremities when they were feeling pain. Aconite levels in drugs and patients' serum after taking daiuzusen were analyzed by liquid chromatography tandem mass spectrometry. Daiuzusen per 1 g of uzu contained aconitine 1.28 μg, mesaconitine 2.31 μg, and hypaconitine 92.89 μg, while jesaconitine was not detected; this was about 5 to 35 times the level of tsumyakushigyakuto per 1 g of uzu. Serum concentrations of hypaconitine peaked in the study at 1.11 ng/mL after about an hour of taking daiuzusen (1 g of uzu). We posit that the immediate effect after taking daiuzusen was due to transmucosal absorption of uzu components. However serum hypaconitine, which we are now able to monitor, is at least one practical way of indicating the use of uzu or bushi containing prescriptions.</p>
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Objective To establish an HPLC method for the simultaneous determination of the 6 aconitum alkaloids in compoundTengwu Ointment.Methods Chromatographic column was Aglient XDB-C18 column (4.6 mm × 250 mm, 5μm), with the mobile phase A of acetonitrile- tetrahydrofuran (25∶8), phase B was 0.1 mol/L ammonium acetate solution (0.5 mL acetic acid added to the 1000 mL) with gradient elution;the flow rate was 1 mL/min;the detection wavelength was set at 235 nm;the column temperature was 25℃.Results Aconitine was in the good linear range of 0.072-0.648μg (r=0.999 5), with the average recovery of 99.29%, RSD=1.25%. Mesaconitine was in the good linear range of 0.062-0.648μg (r=0.999 5), with the average recovery of 99.12%, RSD=0.85%. Hypaconitine was in the good linear range of 0.064-0.384μg (r=0.999 8), with the average recovery of 99.57%, RSD=1.07%. Benzoylaconine was in the good linear range of 0.056-0.672μg (r=0.999 2), with the average recovery of 98.11%, RSD=0.61%. Benzoylmesaconine was in the good linear range of 0.055-0.993μg (r=0.999 9), with the average recovery of 99.27%, RSD=1.10%. Benzoylhypaconine was in the good linear range of 0.078-0.702μg (r=0.999 8), with the average recovery of 99.08%, RSD=1.38%.Conclusion This method is simple, sensitive, repeatable and accurate, which can be used for determination of aconitum alkaloids in compoundTengwu Ointment.
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ObjectiveTo investigate the effects of aconitine, mesaconitine and hypaconitine on calcium release in isolated adult rat cardiac myocytes.MethodsThe left ventricular cardiac myocytes isolated from adult Sprague-Dawley rats were perfused withacnitine, mesaconitine and hypaconitine at 0.3 μmol/L, 1μmol/L, 3 μmol/L for 12 min. The spontaneous calcium release (SCR) rate, the end-diastolic[Ca2+](F0) and the calcium transient amplitude (ΔF) were detected 4 min, 8 min and 12 min after the perfusion. 12 min after the perfusion with acnitine, mesaconitine and hypaconitine at 0.3 μmol/L, the changes of systolic dynamics and calcium transient were detected for the positive inotropic effect. Results Any of aconitine, mesaconitine and hypaconitine induced SCR, mesconitine-induced SCR rate was highest at low concentration (0.3 μmol/L), and aconitine-induced SCR rate highest at high concentration (3 μmol/L). Compared with the control, 12 min after the perfusion with acnitine, mesaconitine and hypaconitine at 3 μmol/L elevated F0 (1.459 ± 0.379, 1.585 ± 0.493, 1.213 ± 0.254vs.1.079 ± 0.108, allP<0.05) and ΔF(1.615 ± 0.455, 2.210 ± 0.756, 1.528 ± 0.422vs. 1.036 ± 0.125, allP<0.05), mesaconitine with ΔF higher than aconitine and hypaconitine. At low concentration (0.3 μmol/L), compared with control, aconitine, mesaconitine and hypaconitine increased ΔF (0.409 ± 0.127, 0.423 ± 0.107, 0.414 ± 0.118vs.0.260 ± 0.065;P<0.05 orP<0.01) and contraction amplitudes (5.464% ± 2.239%, 7.449% ± 2.548%, 5.524% ± 1.645%vs.3.428% ± 0.911%;P<0.05 orP<0.01), prolonged the time to peak of calcium transient (0.041 ± 0.016 s, 0.039 ± 0.009 s, 0.038 ± 0.011 svs.0.032 ± 0.007 s;P<0.05 or P<0.01); compared with aconitine, mesaconitine and hypaconitine decreased calcium transient time constant (0.301 ± 0.054 s, 0.324 ± 0.064 svs.0.361 ± 0.076 s;P<0.05 orP<0.01) and diastolic t50 (0.124 ± 0.035 s, 0.126 ± 0.040 svs.0.157 ± 0.056 s;P<0.05 orP<0.01).ConclusionsAconitine, mesaconitine and hypaconitine reveal the positive inotropic effects couple with the toxic effects. Increased[Ca2+]in cardiac myocytes is the key factor for the positive inotropic effects, but also the risk factor for SCR.
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OBJECTIVE:To study the effects of Heishun tablets combined with Rheum palmatum on the pharmacokinetics of hypaconitine in rats. METHODS:Rats were randomly divided into single drug group(Heishun tablets decoction)and drug combi-nation group(Heishun tablets-R. palmatum mixture decoction),with 18 rats in each group. They were given relevant drugs intragas-trically,by 10 g(medicinal materials)/kg of Heishun tablets. 0.3 ml blood samples were collected before(0 h)and 0.083,0.167, 0.333,0.5,0.75,1,1.5,2,3,4,6,8,10 h after medication with 6 rats at each time point,respectively. The blood concentra-tion of hypaconitine was determined by HPLC-MS using palmatine hydrochloride as internal standard. DAS 2.0.1 software was used to calculate pharmacokinetic parameters. RESULTS:The linear range of hypaconitine was 0.102 4-100 ng/ml (r=0.998 7),and the limit of quantification was 0.1 ng/ml. The pharmacokinetic parameters of single drug group vs. drug combination group were as follows as tmax of (0.50 ± 0.086) h vs. (0.75 ± 0.132)h;t1/2 of (9.967 ± 1.123) h vs. (3.708 ± 0.507) h;AUC0-10 h of (26.087 ± 0.672) μg·h/L vs.(6.516 ± 1.135) μg·h/L;cmax of (6.124 ± 2.312) μg/L vs. (1.592 ± 0.051) μg/L. Compared with single drug group,t1/2,AUC0-10 h and cmax of hypaconitine were decreased in drug combination group,with statistical significance (P<0.05). CONCLUSIONS:R. palmatum can inhibit the absorption of hypaconitine in rats,and speed up the elimination of it in rats.
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Aconitum species were used as the major component in the Chinese and Bhutanese herbal medicines. The species posses many phytochemical compounds which possess many of the pharmacological activities. Diterpene alkaloids were the main compound with the pharmacological activities such as analgesic and against inflammation. This alkaloid possesses certain toxic hydrolyzed bases which could be detoxified by the intervention of recent technologies. Apart from this, the plant possess many alkaloids, amide alkaloids, flavonoids, flavonol glycosides, diterpenoid and norditerpenoid compounds which possess medicinal values. The above mentioned compounds of potent importance were isolated and characterized by the chromatographic separation techniques and their structures were usually elucidated by the spectroscopic studies especially with Nuclear Magnetic Resonance techniques. These compounds were the central target of the medicinal chemist as they possess both medicinal and toxic nature. The measures to be taken in such a way that the medicinal compounds of the plant should be isolated and formulated without the toxic nature. This review encompasses the total phytochemical compounds that have been isolated from various species of the plant genus Aconitum.
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Aim: To study pharmacokinetics of aconitine,mesaconitine and hypaconitine in rats after single oral administration of decoctions composed with Radix Aconiti Lateralis.Methods: Four groups of rats were orally administered four decoctions including decoction a(Sini decoction),decoction b(decoction composed with Radix Aconiti Lateralis),decoction c(decoction composed with Radix Aconiti Laterali and Radix et Rhizoma Glycyrrhizae Praeparata Cum Melle)and decoction d(decoction composed with Radix Aconiti Laterali and Rhizoma Zingiberis),respectively.Quantitative analysis of aconitine,mesaconitine and hypaconitine in rat plasma was achieved using a liquid chromatography-electrospray ionization/tandem mass spectrometry method.Pharmacokinetic parameters were estimated using DAS 2.0.Results: Pharmacokinetic parameters of aconitine,mesaconitine and hypaconitine were different after oral administration of four decoctions according to Radix Aconiti Laterali combined with different herbal medicines.Multiple peaks were observed in plasma concentration-time curve after oral administration of the decoction of herb couple Radix Aconiti Laterali and Radix et Rhizoma Glycyrrhizae Praeparata Cum Melle,and the results showed a delay in tmax and a prolonger in MRT0-t compared with the decoction of Radix Aconiti Latera-lis.When Radix Aconiti Lateralis was combined with Radix et Rhizoma Glycyrrhizae Praeparata Cum Melle and Rhizoma Zingiberis at the same time in Sini decoction,tmax was delayed too but MRT0-t,was shorten than that of the group of Radix Aconiti Lateralis.Conclusion: The pharmacokinetic parameters of the three compounds obtained in this work shows that the pharmacokinetics of aconitine,mesaconitine and hypaconitine were influenced diversely when Radix Aconiti Laterali was combined with different herbal medicines.
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Objective To establish a method for the determination of Alkaloid in cut crude drug of aconite root.Method Contents of aconitine,hypaconitine and mesaconitine incrude drug of aconite root were determined by LC-MS.It was carried out on Inertsil ODS-3 column(150 mm?4.6 mm,5 ?m),with acetonitrile-ammonium acetate buffer solution(60:40) as mobile phase,at a flow rate of 0.2 mL/min and temperature of 30 ℃.Result The method of determining Alkaloid content in crude drug of aconite root was established and 25 lots of crude drugs were determined.Conclusion The method was sensitive and specifical,can be used to control the quality of cut crude drug from aconite root.
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Objective To observe the poisoning components in plasma and histological changes of rabbits with acute toxicity of aconitum kusnezoffii (草乌).Methods Eight rabbits were garaged with aconitum kusnezoffii liquor,aconitum poisoning model was reproduced,electrocardiogram (ECG) and blood pressure were recorded,the concentrations of aconitine,hypaconitine and mesaconitine in plasma after 0.5,1, 2,3 and 6 hours were measured,and the pathological changes of heart,liver and cerebral cortex were observed.Results After garage with poisoning liquor,arrhythmias and the declination of blood pressure, presenting a tendency of progressive aggravation [before garage:(121.98?16.77)/(110.66?8.78) mm Hg, 1 hour after garage:(102.98?8.34)/(90.22?5.85) mm Hg,2 hours after garage:(66.81?9.13)/ (53.40?6.32) mm Hg,1 mm Hg=0.133 kPa,all P
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Objective:To optimize the extraction procedure of total alkaloid in Radix Aconiti.Methods:The orthogonal design(L9(34) ) was carried out for the optimum extraction conditions guided by yield of the extract and the content of aconitine,hypaconitine and mesaconitine determined with RP-HPLC method,the mobile phase was MeOH-H2O2-CHCl3-DEA(70∶30∶ 2∶0.1).Results:The best extraction process was as follows:moisten with ammonia test solution,then leach for 24 hours with 15 times of ether.Conclusion:The optimum extraction methods are rational.
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Aim To develop an HPLC method for the determination of Aconitum alkaloids extracted from Radix aconiti preparata in rats. Methods Waters 2690@996 PAD system was used. The analytical column was a Halsil 100 C18 column (250 mm×4.6 mm ID, 5 μm). The mobile phase was water, methanol and diethyl amine at the ratio of 75∶ 25∶ 0.1. The flow rate was 0.9 mL·min -1. The wavelength of the detector was 240 nm. Results The linear ranges of aconitine in the heart, spleen, lung and kidney were 0.4-100 μg·mL -1, the correlation coefficients were 0.997 2, 0.998 6, 0.999 3 and 0.999 4, respectively. The linear range of aconitine in liver was 2-200 μg·mL -1 and the correlation coefficient was 0.999 0. The linear ranges of hypaconitine in heart, liver, spleen, lung, kidney, brain and spinal cord were 5-100 μg·mL -1, the correlation coefficients were 0.999 4, 0.999 7, 0.999 8, 0.998 4, 0.999 8, 0.999 8 and 0.999 7, respectively. Detection limits (S/N=3) of aconitine and hypaconitine were 0.4 μg·mL -1. The recoveries of aconitine and hypaconitine ranged from 88.7% to 102.2% and 86.5% to 101.3%, respectively, and the RSD of precision of aconitine and hypaconitine was 10%. Conclusion It appears to be an accurate and effective method that can offer reference basis for in toxication of Radix aconiti preparata clinically.
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Objective To establish a method used for determination of Aconitine and Hypaconitine and Mesaconitine in blood by LC/MS/MS.Methods Extraction of blood sample was conducted by use of 1% thirdchloroactic acid and acetonitrile liquid,and the electrospray fog ionic source,and positive ion MRM scan were employed.Result The linearity correlation coefficients(r)was≥0.992 2.The limit of quantification (LOQ S/N=5) was 2.0 ng/ml for Aconitine,0.5 ng/ml for Hypaconitine,and 0.5 ng/ml for Mesaconitine.The recovery of appended contrast in blank blood sample ranged from 91.25% to 103.1% with coefficient of variation (CV,n=6) less than 10.93%.Conclusion The method of LC/MS/MS is sensitive and reliable,and the sample handling is quick and simple,which was suitable for determination of the Aconitine and Hypaconitine and Mesaconitine in blood sample.