Humans ; Aconitine/analysis* ; Chromatography, High Pressure Liquid ; Poisoning/diagnosis*

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 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

To establish a method for the content determination of indexes for measuring aconitic compounds contained in Shenfu injection, in order to provide basis for the evaluation of the curative effect of monkshood in Shenfu injection. The sample were purified and enriched with HF-LPME. ACQUITY UPLC BEH C18 column (2.1 mm x 50 mm, 1.7 microm) was adopted and eluted with a gradient program, with acetonitrile-10 mmol x L(-1) NH4HCO3 (pH 10) as the mobile phases. The flow rate was 0.45 mL x min(-1). The content was determined with ESI and MRM. The results showed that aconitine, hypaconitine and mesaconitine showed a good linear relationship, with r > 0.999, within the range of 0.1-100 ng x L(-1). The recoveries were detected to be 100.1%, 97.4%, 97.5%, with RSD being 1.2%, 1.1%, 1.5%, respectively. This method was used to prove the safety of Shenfu injection, and provide scientific basis for correct evaluation of curative effect of monkshood, as well as a reliable, simple and practical means for quality control of monkshood-containing Chinese materia medica preparations.
Aconitine ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Mass Spectrometry ; methods ; Quality Control

OBJECTIVETo set up a capillary electrophoresis method with field-amplified sample injection for the determination f aconitine and hypaconitine in Gucixiaotong Ye.

METHODAn uncoated fused-silica capillary column (50 microm x 50 cm, effective length 42 cm) was used as the separation channel. The running buffer was 50 mmol x L(-1) phosphate electrolyte solution (pH 9)-m nol (90:10) , the running voltage was 10 kV and the capillary inlet was dipped in methanol for 5 s prior to electrokinetic injection (12 kV, 30 s), the detection wavelength was set at 235 nm.

RESULTAconitine and hypaconitine were linear in the concentration ranges of 17.2-275 microg x L(-1) and 34. 4-550 microg x L(-1), respectively. The average recovery was more than 93.9% with the RSD of 3.8%. This method could enrich 500 fold of aconitine alkaloid.

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

Aconitine ; analogs & derivatives ; analysis ; isolation & purification ; Buffers ; Electricity ; Electrophoresis, Capillary ; methods ; Hydrogen-Ion Concentration ; Injections ; Solutions ; Time Factors

In this study, we explored the rationality of processing methods and mechanism of Aconiti Lateralis Radix (Fuzi) through comparing the chemical contents of diester alkaloids (DAs) and monoester alkaloids (MAs) in the raw material of Fuzi and its processed products. The results showed that the toxicity potency of MAs is at least lower than 1/64 to 1/180 of the toxicity potency of DAs. The contents of DAs in processed Fuzi decreased to 1/76.5 to 1/38.3 of the value of raw Fuzi. The contents of MAs in processed Fuzi significantly increased by 4.6 to 5.2 fold or basically the same as that of the raw Fuzi. The values of MAs/DAs of processed Fuzi were enhanced by 30 to 390 fold of the raw Fuzi. It was found that the contents of DAs were insignificantly different between "Wu dan fu pian" (steaming or stir-frying without Danba) and "Dan fu pian" (steaming or stir-frying with Danba). The result suggested that the abilities of "eliminating toxicity" of different processing methods were equivalent at all. In contrast, the contents of MAs contained in "Wu dan fu pian" were of 5.3 to 8.7 fold higher than the values in "Dan fu pian". This result suggested the processing method by steaming or stir-frying without Danba might have better effect for "conserving property" than the method processed with Danba stipulated by China Pharmacopoeia. We believe that the new processing method without Danba can be recommended in further application due to it offers a simple procedure and it will not introduce inorganic impurities in the products.
Aconitine ; analogs & derivatives ; analysis ; toxicity ; Aconitum ; chemistry ; Animals ; Chromatography, High Pressure Liquid ; Cluster Analysis ; Drugs, Chinese Herbal ; analysis ; toxicity ; Male ; Rats ; Rats, Sprague-Dawley ; Technology, Pharmaceutical ; methods

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.
Aconitine ; analysis ; Aconitum ; chemistry ; Chromatography, High Pressure Liquid ; Diterpene Alkaloids ; analysis ; Drugs, Chinese Herbal ; chemistry ; Phytochemicals ; analysis ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry

AIMTo separate and quantitatively determine six alkaloids: aconitine, mesaconitine, hypaconitine, beiwutine, benzoylaconine and benzoylmesaconine in the Chinese traditional medicine Radix Aconiti Lateralis Preparata (Fuzi).

METHODSA RP-ion-pair HPLC method was established. An AichromBond-1 C18 column was used at a column-temperature of 35 degrees C. The mobile phase was CH3CN5 mmol x L(-1) NaH2PO4(50:50) containing 7 mmol x L(-1) SDS at a flow-rate of 1.0 mL x min(-1). The detector was set at UV 235 nm.

RESULTSThese six alkaloids can be completely separated and determined quantitatively.

CONCLUSIONThis method is accurate and suitable for the determination of six alkaloids in Fuzi.

Aconitine ; analogs & derivatives ; analysis ; isolation & purification ; Aconitum ; chemistry ; Alkaloids ; analysis ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Plant Roots ; chemistry ; Plants, Medicinal ; 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 establish an HPLC method for the determination of three kind of diester diterpenoid alkaloids (DDAs) in Aconitum carmichaeli and its processed products namely mesaconitine, aconitine and hypaconitine.

METHODA Zorbax Eclipse XDB C18 column was used, and ammonium acetate solution-acetonitrile as the mobile phase, with a flow rate of 1.0 mL x min(-1) and a detection wavelength of 230 nm.

RESULTMesaconitine, aconitine and hypaconitine, were separated, and the calibration curves of them were in good linearity over the range of 0.035 7-1.784 microg (r = 0.9999), 0.0126-0.632 microg (r = 0.9997) and 0.0334-1.672 microg (r = 0.9997) respectively.

CONCLUSIONThis method is simple and accurate. It can be used in the identification and quality control of A. carmichaeli.

Aconitine ; analogs & derivatives ; analysis ; Aconitum ; chemistry ; Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; methods ; Hot Temperature ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Technology, Pharmaceutical ; methods