Identification and Characterization of Sclerotium delphinii Causing Southern Blight on Aconitum kusnezoffii in Northeast China.

Aconitum kusnezoffii is a perennial medicinal plant that belongs to the Ranunculaceae family and is distributed mainly in Northeast and North China. In July 2018, a typical southern blight disease of A. kusnezoffii was observed in commercial fields of Qingyuan County, Fushun City, Liaoning Province, China. The fungus mainly infected stem base and tuberous roots of the plant by wrapping the hyphae and absorbing nutrition, resulting in tuberous root wilted or whole plant death. Morphological characteristics of colony and sclerotia of three representative strains isolated from the diseased plants differed from those of Sclerotium rolfsii isolated from A. carmichaelii. Sclerotia were large (0.8 to 5.1 mm), reddish-brown, and irregular and had pitted surfaces, and the hyphae were white, compact, or fluffy, with a growth rate ranging from 8.0 to 10.1 mm/day. Phylogenetic analysis of the internal transcribed spacer and the large subunit sequences of Akln6, Akln9, and Akln15 showed that three strains isolated from A. kusnezoffii formed a unique and well-supported clade that groups with the reference isolates of S. delphinii. Based on phylogenetic analysis and cultural and morphological characteristics, the three isolates of A. kusnezoffii were identified as S. delphinii. The optimum temperature for mycelial growth of the three tested isolates was 30°C, and sclerotia formed and matured more easily at 20°C. Light promoted the growth of mycelial, whereas dark was beneficial to the formation and maturation of sclerotia. The pathogenicity of S. delphinii showed stronger than S. rolfsii at low temperature (20°C). This is the first report of S. delphinii causing southern blight on A. kusnezoffii in China, and this finding provides a basis for disease-accurate diagnosis and the development of effective management strategies.

[A case of aconitum kusnezoffii intoxication with severe arrhythmia].

Aconitum kusnezoffii is a traditional Chinese medicine of Ranunculaceae family. Its toxicity is relatively strong, and its dosage is similar to that of poisoning. In clinical practice, poisoning events are often caused by excessive dosage or improper use. There is no specific antidote for kusnezoff root poisoning. Severe kusnezoff root poisoning can cause malignant arrhythmia and even death.A case of severe kusnezoff monkshood poisoning was reported in January 2021, which was treated with nificaran hydrochloride for injection in the emergency medicine department of the First Hospital of Handan City. The patient developed ventricular tachycardia, ventricular fibrillation and AS syndrome. In addition to conventional treatment, the patient did not have arrhythmia again after intravenous injection of 25 mg of nifekalan load and continuous pumping of 0.4 mg/kg/h for 7 hours, and did not relapse after discontinuation of nifekalan 24 hours later. It is suggested that the malignant arrhythmia caused by clinical severe kusnezoff monkshood poisoning can be controlled by nifekalan. Whether nifekalan is superior to conventional antiarrhythmic drugs still needs more accumulation and verification of clinical application data.

Classification of diterpenoid alkaloids from Aconitum kusnezoffii Reichb. by liquid chromatography-tandem mass spectrometry-based on molecular networking.

Trace amounts of components in traditional Chinese medicine are considered pharmacological active substances used for treating many serious diseases. However, purifying all the trace substances and making clear their structures are not easy. In this context, high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry based molecular networking was applied to investigate the chemical constituents of the roots of Aconitum kusnezoffii Reichb., which led to the identification of 33 nodes in different groups (N1-N33). Based on the excremental fragmentation pathway of known diterpenoid alkaloids (1-9) and comparisons of characteristic ions and characteristic loss of analogs in literature, the structures of unknown ions were deduced. This work lays a foundation for the evaluation of the clinical basis and mechanism of traditional Chinese medicine from the aspects of chemistry. In this paper, the method speculation of unknown natural products by means of molecular network method is expected to be applied in the discovery and change law of relevant active components in clinical pharmacology and the change of complex systems caused by trace active compounds.

Study on difference of transitional components in blood of Radix Aconiti processed by Hezi decoction / 中国药理学通报

Aim To analyze the differences of transitional components in serum before and after processing of Radix Aconiti, and to reveal the processing mechanism of Radix Aconiti from the perspective of serum pharmacochemistry. Methods Radix Aconiti prepared by processing Radix Aconiti and Hezi decoction was subdivided into 0.5% sodium carboxymethylcellulose suspension, and then the drug containing serum was prepared. The UPLC characteristic maps of serum in blank group, Radix Aconiti group and Radix Aconiti prepared by processing Radix Aconiti Decoction were established, analyzed and compared, and the differences of serum migration components before and after processing were determined. Results A total of 11 blood transitional components were detected and 6 prototype components were identified in the drug containing plasma samples of raw Aconitum group. A total of 14 blood transitional components were detected and 8 prototype components were identified in the drug containing plasma samples of processed Aconitum group of Hezi decoction. Conclusions There are significant differences in serum transitional components before and after processing, which provides an experimental basis for the further study of the processing mechanism of Herba Aconiti in Hezi Decoction in the future.

The complete mitochondrial genome of Aconitum kusnezoffii Rchb. (Ranales, Ranunculaceae).

Aconitum kusnezoffii Rchb. is a medicinal plant in the Ranunculaceae family. In this study, we report the first complete mitochondrial genome of A. kusnezoffii. The total length of the mitochondrial genome of A. kusnezoffii is 440,720 bp and the GC content of 46.85%. The mitochondrial genome contained 37 protein-coding genes, 29 tRNAs, and three rRNAs. These data will provide the basis for the systematic evolutionary analysis of Ranunculaceae.

Quality tracing evaluation strategies of compatible materials in Aconitum proprietary Chinese medicines.

The proprietary Chinese medicine (PCM) has become a significant supplement of modern medicine. Nevertheless, the absence of quality control standard of compatible materials in PCM has led to serious adulteration, which has an extremely bad effect on safety of drug use and clinical efficacy. Here, a quality tracing evaluation strategy of compatible materials in 32 Aconitum proprietary Chinese medicines (APCMs) was established, including data normalization, model development, model verification, and unknown prescription cracking. The model was delimited based on the weighted content of total 9 key alkaloids in 24 APCMs, which were 5.65-57.10 µg/g for extract medicines and 42.62-380.61 µg/g for powder medicines. Three newly published commercial APCMs, including Wangbi Tablet, Wangbi Granule, and Fengshigutong Capsule, were used to verify its reliability and the results proved to be positive. Moreover, a novel prescription cracking approach was proposed to decode the content of each material in five unknown prescriptions including Yaoxitong Capsule, Tongrendahuoluo Pill, Xinbao Pill, Dahuoluo Capsule, and Mugua Pill. Ultimately, the single or two compatible Aconitum materials in APCMs was successfully decoded and the processed level of the materials were effectively judged. This study for the first time established a practical strategy for supervision and cracking of compatible materials in PCMs and is of great significance to improve the quality control of PCMs.

Resource allocation strategies among vegetative growth, sexual reproduction, asexual reproduction and defense during growing season of Aconitum kusnezoffii Reichb.

Natural plants must actively allocate their limited resources for survival and reproduction. Although vegetative growth, sexual reproduction, asexual reproduction and defense are all basic processes in the life cycle of plants, the strategies used to allocate resources between these processes are poorly understood. These processes are conspicuous in naturally grown Aconitum kusnezoffii Reichb., which makes it a suitable study subject. Here, the morphology, dry matter, total organic carbon, total nitrogen and aconitum alkaloid levels of shoot, principal root (PR) and lateral roots were measured throughout the growing season. Then, transcriptome and metabolite content analyses were performed. We found that vegetative growth began first. After vegetative growth ceased, sexual development began. Flower organ development was accompanied by increased photosynthesis and the PR consumed temporarily stored resources after flower formation. Asexual propagule development initiated earlier than sexual reproduction and kept accumulating resources after that. Development was slow before flower formation, mainly manifesting as increasing length; then, after flower formation it accelerated via enhanced material transport and accumulation. Defense compounds were maintained at low levels before flowering. In particular, the turnover of defense compounds was enhanced before and after flower bud emergence, providing resources for other processes. After flower formation, defense compounds were accumulated. The pattern found herein provides a vivid example for further studies on resource allocation strategies. The exciting finding that the PR, as a more direct storage site for photosynthate, is a buffer unit for resources, and that defense compounds can be reused for other processes, suggests a need to explore potential mechanisms.

Kusnezosines A-C, three C19-diterpenoid alkaloids with a new skeleton from Aconitum kusnezoffii Reichb. var. gibbiferum.

Kusnezosines A-C (1-3), three C19-diterpenoid alkaloids with a new skeleton which featured an undescribed lactone type D-ring, were isolated from the roots of Aconitum kusnezoffii Reichb. var. gibbiferum. Kusnezosines A-C are the first naturally occurred C19-diterpenoid alkaloids which possessing an unprecedented lactone D ring, this structure was formed by the cleavage of bond between C-15 and C-16 and a successive lactonization. Their structures were established on the basis of comprehensive spectroscopic data analysis. Besides, another 12 known ones were isolated from this plant, analgesic activity tests on the isolated alkaloids were also carried out.

Quality evaluation of Xiaojin Pills based on antiplatelet aggregation biological potency assay / 中草药

Objective: To establish a biological potency assay for Xiaojin Pills against platelet aggregation in vitro, evaluate the quality consistency of Xiaojin Pills, and screen traditioanal Chinese medicines which play the role of promoting blood circulation in Xiaojin Pills. Methods: Xiaojin Pills and ten Chinese medicines [artificial musk, Momordica cochinchinensis, Aconitum kusnezoffii, Liquidambar formosana, Boswellia carterii, Commiphora myrrha, Faeces Trogopterori, Angelica sinensis, Pheretima aspergillum, Fragrant Ink] in its formula were extracted by ultrasound in 40% methanol. The antiplatelet aggregation rate of the extract was measured by platelet aggregation meter. The platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were prepared from abdominal aorta of rats. The platelet aggregation was induced by adenosine diphosphate (ADP). With sodium ferulate as a standard reference material, the biological potency of antiplatelet aggregation of Xiaojin Pills was calculated by the simplified probit principle. Results: The results showed that the biological potency of Xiaojin Pills was between 0.598 and 1.338 U/mg among different manufacturers and batches. In Xiaojin Pills group, Pheretima, Faeces Trogopterori, and Momordicae Semen had stronger inhibitory effects on platelet aggregation with inhibition rates of 70.87%, 31.83% and 67.52%, respectively. Conclusion: The quality consistency of Xiaojin Pills from different manufacturers and batches is poor, and Pheretima, Faeces Trogopterori, and Momordicae Semen may be the key drugs for Xiaojin Pills to play the role of promoting blood circulation.

[Study on absorbed components of Aconitum kusnezoffii under Yunnan Baiyao compatibility in effect of activating blood circulation and removing blood stasis].

Rat model of blood stasis syndrome was prepared by subcutaneous injecting of epinephrine hydrochlorid,then the model rats were administrated by Yunnan Baiyao for 15 days. Blood rheology,coagulation function and histopathology were chosen as indicators to evaluate the successful replication of blood stasis syndrome model and the treatment effect of Yunnan Baiyao. UPLC-Q-TOF-MS was used to rapidly analyze the serum samples of blood stasis syndrome rat after 15 days Yunnan Baiyao treatment,Progenesis QI software was employed to identify the alkaloids components. The results showed that Yunnan Baiyao reduced the plasma viscosity and whole blood viscosity of rats with blood stasis syndrome,prolonged thrombin and prothrombin time,reduced fibrinogen content,and effectively improved pathological state such as inflammatory cell infiltration,blood stasis,congestion and edema of various organs in rats with blood stasis syndrome. Seven alkaloids components from Aconitum kusnezoffii,including karacolidine,senbusine B,isotalatizidine,karakoline,denudatine,talatisamine and chasmanine were found in the rat serum after Yunnan Baiyao treatment. Based on the effectiveness of Yunnan Baiyao in the treatment of blood stasis syndrome induced by epinephrine hydrochloride in rats,alkaloids components from the root of A. kusnezoffii absorbed into blood after Yunnan Baiyao treatment were clarified rapidly and accurately with the help of UPLC-Q-TOF-MS. Karacolidine,senbusine B,isotalatizidine,karakoline,denudatine,talatisamine and chasmanine are the pharmacodynamic material basis of the root of A. kusnezoffii for activating blood circulation and removing blood stasis.

Metabolomics of Clinical Poisoning by Aconitum Alkaloids Using Derivatization LC-MS.

The root of Aconitum kusnezoffii (Caowu in Chinese, CW) is not only commonly used as a traditional Chinese medicine (TCM), but also served as a tonic in China. Due to its high toxicity, clinical poisoning cases induced by CW have frequently been reported. However, the mechanism is still unclear. In this study, Aconitum alkaloids and altered endogenous metabolites in CW poisoning patients were investigated to elucidate the possible intoxication mechanism. Eighteen alkaloids, including 6 toxic diester diterpenoid alkaloids (DDAs), were determined from the sera of patients. At the same time, 5-(diisopropylamino)amylamine (DIAAA) derivatization-ultrahigh performance liquid chromatography- quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF/MS) approach was applied in the metabolomics analysis to find much more carboxyl-containing metabolites (CCMs), which are the essential components for life and critical to elucidate the mechanism of toxicity. As a result, 32 altered metabolites after poisoning were identified. Among them, hydroxyeicosatetraenoic acids (HETEs) and some dicarboxylic acids were first found to be related to Aconitum alkaloids toxicity. Finally, biological pathway analysis indicated that the significantly changed metabolites were primarily involved in amino acid metabolism, TCA cycle, fatty acid metabolism, pyruvate metabolism, arachidonic acid metabolism, sphingolipid metabolism and so on. These results can not only provide more information on the mechanism of CW intoxication but also help the clinical diagnosis of CW poisoning.

Study on Compatibility Proportion of Mongolian Medicine “Terminalia chebula Decomposing Poison of Aconitum kusnezoffii ” / 中国药房

OBJECTIVE： To investigate the best compatibility proportion of Mongolian medicine “Terminalia chebula decomposing the poison of Aconitum kusnezoffii”. METHODS： Totally 40 rats were randomly divided into blank control group （0.05％ CMC-Na）， raw A. kusnezoffii group （0.12 g/kg by crude drug） and raw A. kusnezoffii-T. chebula  （1 ∶ 3，1 ∶ 1，3 ∶ 1， mass ratio） group （0.12 g/kg raw A. kusnezoffii by raw material）， 8 rats in each group. They were given relevant medicine intragastrically once a day， for consecutive 28 d. 0.5 h after last medication， serum contents of cTn-I and MB were determined， and the changes of cardiological structure were observed； the detoxification effects of T. chebula on cardiotoxicity induced by A. kusnezoffii were investigated. Binding rates of 3 kinds of aconitine （concentrations of aconitine， mesaconitine and hypaconitine were 5， 10， 20， 40， 80， 160， 400 ng/mL） to binding rate of plasma protein with normal human plasma were determined by equilibrium dialysis combined with liquid chromtography-mass spectrometry. The concentrations of 3 kinds of aconitine were fixed as 100 ng/mL. After adding main detoxification component tannic acid in different proportions of T. chebula （1 ∶ 0.025， 1 ∶ 0.075， 1 ∶ 0.1， 1 ∶ 0.5）， the effects of them on plasma protein binding rate of 3 kinds of aconitine were investigated； the possible mechanism  of T. chebula decomposing the poison of A. kusnezoffii inducing cardiotoxicity were investigated. RESULTS： In detoxification experiment， compared with blank control group， serum contents of cTn-I and MB were increased significantly in raw A. kusnezoffii group （P＜0.05）. There were obvious pathological changes in myocardial tissue， such as disorder of cell arrangement， cell shrinkage and cytoplasm staining. Compared with raw A. kusnezoffii group， serum contents of cTn-I and MB in raw A. kusnezoffii-T. chebula （1 ∶ 3， 1 ∶ 1， 3 ∶ 1） groups were decreased significantly （P＜0.05）， and there was no significant difference between the raw A. kusnezoffii-T. chebula （3 ∶ 1， 1 ∶ 1） groups and blank control group （P＞0.05）； myocardial pathological changes were improved to varying degrees. The structure of myocardial tissue in raw A. kusnezoffii-T. chebula （3 ∶ 1） groups were similar to blank control group. In the mechanism investigation experiment under the condition of different concentrations， plasma protein binding rates of 3 kinds of aconitine with normal human plasma were about 30％， without statistical significance （P＞0.05） and significant concentration-dependent manner. After adding tannic acid， plasma protein binding rate of 3 kinds of aconitine in A. kusnezoffii was decreased to different extent； when 3 kinds of aconitine were combined with tannic acid in the ratio of 1 ∶ 0.1， 1 ∶ 0.075 and 1 ∶ 0.5， the plasma protein binding rate decreased significantly （P＜0.05）， in proportion-dependent manner. CONCLUSIONS： Compatible use of raw A. kusnezoffii-T. chebula （3 ∶ 1） show that best detoxification effect on A. kusnezoffii-induced cardiotoxicity. Under this compatibility ratio， the plasma protein binding rate of 3 kinds of aconitine in A. kusnezoffii with normal human plasma is relatively high and the free drug concentration is relatively low.

Study on absorbed components of Aconitum kusnezoffii under Yunnan Baiyao compatibility in effect of activating blood circulation and removing blood stasis / 中国中药杂志

Rat model of blood stasis syndrome was prepared by subcutaneous injecting of epinephrine hydrochlorid,then the model rats were administrated by Yunnan Baiyao for 15 days. Blood rheology,coagulation function and histopathology were chosen as indicators to evaluate the successful replication of blood stasis syndrome model and the treatment effect of Yunnan Baiyao. UPLC-Q-TOF-MS was used to rapidly analyze the serum samples of blood stasis syndrome rat after 15 days Yunnan Baiyao treatment,Progenesis QI software was employed to identify the alkaloids components. The results showed that Yunnan Baiyao reduced the plasma viscosity and whole blood viscosity of rats with blood stasis syndrome,prolonged thrombin and prothrombin time,reduced fibrinogen content,and effectively improved pathological state such as inflammatory cell infiltration,blood stasis,congestion and edema of various organs in rats with blood stasis syndrome. Seven alkaloids components from Aconitum kusnezoffii,including karacolidine,senbusine B,isotalatizidine,karakoline,denudatine,talatisamine and chasmanine were found in the rat serum after Yunnan Baiyao treatment. Based on the effectiveness of Yunnan Baiyao in the treatment of blood stasis syndrome induced by epinephrine hydrochloride in rats,alkaloids components from the root of A. kusnezoffii absorbed into blood after Yunnan Baiyao treatment were clarified rapidly and accurately with the help of UPLC-Q-TOF-MS. Karacolidine,senbusine B,isotalatizidine,karakoline,denudatine,talatisamine and chasmanine are the pharmacodynamic material basis of the root of A. kusnezoffii for activating blood circulation and removing blood stasis.

[Content determination of four diester diterpenoid alkaloids in leaves of Aconitum kusnezoffii by HPLC].

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.

Content determination of four diester diterpenoid alkaloids in leaves of Aconitum kusnezoffii by HPLC / 中国中药杂志

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.

Characterization of 19 microsatellite loci in the clonal monkshood Aconitum kusnezoffii (Ranunculaceae).

PREMISE OF THE STUDY: Microsatellite loci were isolated and characterized from Aconitum kusnezoffii (Ranunculaceae) to estimate male and female reproductive success and evaluate the effects of clonal growth on sexual reproduction. METHODS AND RESULTS: A genomic enrichment approach was used to develop microsatellite markers. In three investigated A. kusnezoffii populations, a total of 19 microsatellite loci were successfully amplified, and 13 of these loci were polymorphic. Most of the primer pairs designed for the identified loci also amplified corresponding microsatellite loci in A. barbatum var. puberulum and A. alboviolaceum. CONCLUSIONS: The identified microsatellite loci will be useful for quantifying male and female fitness in A. kusnezoffii and evaluating the effects of clonal growth on sexual reproduction.

An Efficient High-performance Liquid Chromatography Combined with Electrospray Ionization Tandem Mass Spectrometry Method to Elaborate the Changes of Components Between the Raw and Processed Radix Aconitum kusnezoffii.

BACKGROUND: Crude radix Aconitum kusnezoffii (RAK) has great toxicity. Traditional Chinese medicine practice proved that processing may decrease its toxicity. In our previous study, we had established a new method of RAK processing (Paozhi). However, the mechanism is yet not perfect. OBJECTIVE: To explore the related mechanism of processing through comparing the chemical contents. MATERIALS AND METHODS: A new processing method of RAK named stoving (Hong Zhi) was used. In particular, RAK was stored at 110°C for 8 h, and then high performance liquid chromatography combined with electrospray ionization tandem mass spectrometry (HPLC-ESI-MS(n)) was developed for the detection of the alkaloids of the crude and processed RAK decoction pieces. RESULTS: Thirty components of the crude RAK were discovered, among which, 23 alkaloids were identified. Meanwhile, 23 ingredients were detected in the processed RAK decoction pieces, among which, 20 alkaloids were determined yet. By comparison, eight alkaloids were found in both crude and processed RAK decoction pieces, 15 alkaloids were not found in the crude RAK, however, 10 new constituents yield after processing, which are 10-OH-hypaconine, 10-OH-mesaconine, isomer of bullatine A, 14-benzoyl-10-OH-mesaconine, 14-benzoyl-10-OH-aconine, 14-benzoyl-10-OH-hypaconine, dehydrated aconitine, 14-benzoylaconine, chuanfumine, dehydrated mesaconitine. CONCLUSION: The present study showed that significant change of alkaloids was detected in RAK before and after processing. Among them, the highly toxic diester alkaloids decreased and the less toxic monoester alkaloids increased. Moreover, the concentration changes significantly. HPLC-ESI-MS(n) are Efficient to elaborate the mechanism of reduction of toxicity and enhancement efficacy after processing. SUMMARY: Stoving is a simple and effective method for the processing of radix Aconitum kusnezoffii.In the positive mode, the characteristic fragmentations of Aconitum alkaloids were obtained.The highly toxic alkaloids have decreased, and the new constituents appeared, which has explained successfully the processing mechanism of radix Aconitum kusnezoffii in chemistry.

Analysis of Different Parts of Aconitum kusnezoffii Reichb.Using Fourier Transform Infrared Spectroscopy / 世界科学技术-中医药现代化

The radix,leaf,flower and bud of raw medicinal materials and extraction of total alkaloids of Aconitum kusnezoffii Reichb.were all involved in this investigation.All the compositions from the samples were analyzed through fourier transform infrared spectroscopy (FTIR) combined with second derivative IR spectroscopy and two-dimensional IR correlation spectroscopy (2D-IR).It was found that the spectra of raw medicinal materials showed that the radix of A.kusnezoffii Reichb.featuring a large quantity of starch was the same as starch with the characteristic peaks at 1,155,1,070 and 1,019.The leaf,flower and bud contained the similar aromatic hydrocarbons (1,600),glycosides (1,050-1,070),while lipids were not clear.The characteristic peaks of the buds,flowers and leaves were all at 1,595 cm-1 (vibration of phenyl framework) and 1,262 cm-1 (=C-O).Therefore,it was suggested that the common compound of the three parts be diterpenoid alkaloids.Second derivative IR spectroscopy showed that the characteristic peaks of radix was stronger than those of the flower,leaf and bud at 1,712 cm-1 (C=O),which proved that the quantity of characteristic peaks in the radix was larger than those in the flower,leaf and bud.In addition,six autopeaks at 1,745,1,650,1,560 (the most strong),1,465,1,400,1,300 were detected from the radix.The similar autopeaks at 1,745,1,650,1,560 (the most strong),1,465,1,400,1,300 were found in the leaf,bud and flower.In conclusion,it was demonstrated that the macro-fingerprint infrared spectroscopic identification method provided a large quantity of the comprehensive information and entirely grasped the quality of A.kusnezoffii Reichb.Besides,FTIR and 2D-IR provided massive information of the integral structures of the radix,leaf,flower and bud of A.kusnezoffii Reichb.and verified the differences between the four parts of the herb in physical structure and the contents,laying a foundation for further systematic work.

Study on Purification Technology of Total Alkaloid from the Flos of Aconitum kusnezoffii / 中国药房

OBJECTIVE:To optimize the purification technology of total alkaloid from the flos of Aconitum kusnezoffii. METH-ODS:The content of total alkaloid from the flos of A. kusnezoffii was determined by acid-base titration. The purification technology of total alkaloid from the flos of A. kusnezoffii was optimized by ion resin with resin type,mass concentration of loading liquid and exchange speed as factors,maximum adsorption quantity,desorption rate and mass fraction of total alkaloid as index,and verifica-tion test was conducted. RESULTS:The optimal purification technology was as follows as type 732 cation exchange resin,mass concentration of loading liquid 0.32 g/L,exchange speed of 7 column volume(BV)/h. In validation test,the content of total alka-loid was 86.88%(RSD=0.52%,n=3),and desorption rate was 92.81%(RSD=0.40%,n=3)averagely. The extraction trans-port rate of total alkaloid from 3 batches of the flos of A. kusnezoffii was 81.76% and purification transport rate was 89.47% in av-erage. CONCLUSIONS:The established method is stable and feasible,and shows high transport rate.

Study of Single-dose Toxicity of Aconitum Kusnezoffii Reichb. Pharmacopuncture in Rats.

OBJECTIVE: This study was performed to analyze the singledose toxicity of Aconitum kusnezoffii Reichb. pharmacopuncture (AKRP). METHODS: All experiments were conducted at the Korea Testing & Research Institute (KTRI), an institute authorized to perform non-clinical studies, under the regulations of Good Laboratory Practice (GLP). Twenty (20) Sprague-Dawley rats were chosen for the pilot study. The animals were divided into four groups of five animals per group: group 1 (G1) being the control group with each animal receiving an injection of 0.3 ml of saline and groups 2, 3, and 4 (G2, G3, and G4) being the experimental groups with each animal receiving an injection of 0.1, 0.2 or 0.3 ml of AKRP, respectively. This study was conducted with the approval of the Institutional Animal Ethics Committee. RESULTS: No deaths occurred in any of the 4 groups, and the LD50 of AKRP administered via IV was higher than 1.77 ml/kg. Some changes in the weights of the male rates were observed between the control group and the experimental groups, but no significant differences were noted in the weights of the female rats. To check for abnormalities in organs and tissues, we stained representative sections of each specified organ with Hematoxylin & Eosin for light microscopic examination. The results showed no significant differences in any of the organs or tissues. CONCLUSIONS: The above findings suggest that Aconitum kusnezoffii Reichb. pharmacopuncture is a relatively safe treatment. Further studies on the subject should be conducted to yield more concrete evidence.