一测多评法测定乌头类药材中生物碱

目的 建立一测多评法同时定量测定乌头类药材中6种乌头类生物碱.方法 采用高效液相色谱法同时测定乌头类药材中6种生物碱,以乌头碱为内参物,测定其与次乌头碱、新乌头碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、苯甲酰新乌头原碱的相对校正因子,利用乌头碱和相对校正因子对其他5种生物碱成分进行测定,同时利用外标法测定这6种成分,比较两种测定方法的差异,验证一测多评法的可行性和准确性.结果 在一定线性范围内,乌头碱与次乌头碱、新乌头碱、苯甲酰乌头原碱、苯甲酰次乌头原碱和苯甲酰新乌头原碱的相对校正因子分别为0.780、1.008、0.836、0.907、0.987,在不同实验条件下相对校正因子重现性良好,不同乌头类药材中6种生物碱成分含量计算值与实测值间无显著性差异.结论 一测多评法在乌头类药材及制剂质量控制中应用是可行的、准确的.     

Investigation of the diterpene alkaloids of Aconitum species native to the Carpathian Basin

Aconitum species accumulate highly toxic diterpene alkaloids, which have attracted considerable interest because of their complex structure, interesting chemistry and noteworthy physiological effects. The authors carried out phytochemical analysis of two Aconitum species native to the Carpathian Basin, aiming the isolation of diterpene alkaloids. The multistep chromatographic purification of the extracts of A. vulparia whole plant and A. toxicum root resulted in the isolation of 16 alkaloids. Structure elucidation by means of spectroscopic investigations revealed the presence of 3 (acovulparine, vulparine, acotoxinine) new and 13 already known compounds (anthranoyllycoctonine, delectinine, delcosine, finetiadine, lycoctonine, N-methyl-N-deethyllycocotine, septentriodine, aconitine, aconosine, dolaconine, neoline, songorine, songo-ramine). Alkaloids isolated from A. vulparia are C19 diterpene alkaloids possessing aconitane skeleton, whilst from A. toxicum two C20 diterpene alkaloids with napelline and five diterpene alkaloids with aconitane skeleton were identified. The pharmacological testing of the majority of the isolated compounds has not been reported in the literature yet. Since some of the diterpene alkaloids are structurally very similar to compounds which were active in different in vitro or in vivo experiments, the pharmacological testing of the alkaloids reported in this article seems to be reasonable. The investigation of the compounds may help to reveal structure-activity relationships as well.     

Separation and identification of Aconitum alkaloids and their metabolites in human urine.

To study the safety of Aconitum medicinal herbs in clinic and identify Aconitum alkaloids poisoning in forensic medicine, Aconitum alkaloids and their metabolites were separated and identified in human urine by liquid chromatography-electrospray ionization-multi-stage mass spectrometry (LC-ESI-MS(n)) and chemical pathway of metabolism was investigated. The alkaloids and their metabolites in the urine sample were extracted with solid-phase cartridges and separated by HPLC with acetonitrile-water-formic acid (40:60:0.5) mobile phase. Structures of five metabolites and three parent Aconitum alkaloids were identified with multi-stage mass spectrometry data through comparison with authentic substances as aconitine (M(1)), mesaconitine (M(2)), hypaconitine (M(3)), benzoylaconine (M(4)), benzoylmesaconine (M(5)), benzoylhypaconine (M(6)), 16-O-demethylaconitine (M(7)) and 16-O-demethylhypaconitine (M(8)), respectively. Among them, M(8) was identified and reported for the first time. Metabolic pathways of Aconitum alkaloids in human body were proposed.     

Alkaloidal constituents fromAconitum jaluense

Aconitum jaluense Komar. (Ranunculaceae) is one of the Aconitum plants growing in Korean peninsula. An investigation of the alkaloidal constituents of this species led to the isolation of seven C19-norditerpenoid and a C20-diterpenoid alkaloid. Three of them have been identified as neoline, mesaconitine, and hypaconitine, which were isolated from this plant collected from Mt. Bultasan in the north part. The other five alkaloids were determined as lipomesaconitine, lipohypaconitine, 15alpha-hydroxyneoline, hokbusine A, and napelline, which have not been found in this plant. Structures of those alkaloids were determined on the basis of their spectral data. It is of interest to note that a comparison of the present work and the previous report showed some differences in the alkaloidal contents.     

草乌花及其煎煮液中二萜生物碱的电喷雾串联质谱研究草乌花及其煎煮液中二萜生物碱的电喷雾串联质谱研究

目的 对草乌花及其煎煮液中的二萜生物碱进行定性分析,说明煎煮前后化学成分发生的变化。 方法用注射泵自动进样,电喷雾离子阱串联质谱直接分析草乌花及其煎煮液中生物碱混合物。结果 在生草乌花中发现3个新生物碱,草乌花煎煮后其中的双酯型生物碱和三酯型生物碱都发生水解,前者水解为苯甲酰乌头原碱和乌头原碱类生物碱,后者水解为3-乙酰-乌头原碱类生物碱。结论 该法简便、快速、灵敏、特异性强,为乌头属植物煎煮液中的生物碱分析提供了新途径。     

Determination of Aconitine and its Metabolites in Animal Experiment by LC／ESI—MS^n

Aconitumspecies (Ranunculaceae) ,whichdistributewidelyinmiddleandlowerreachesoftheYangziRiver ,northandsouthwestofChina ,containhighlytoxicdiesterditerpeneAconitumalkaloids ,especiallyhighcontentofaconitine .Aconitine ,withanLDforhumanof 3- 5mg ,issometimesusedinsuicidesandhomicidesbecauseofitshightoxicity .Forsearchasmallamountofthetoxicant,itsidentificationisverydifficult.Thus ,forclinicalandforensicmedicine ,amethodforsensitivedeterminationandidentificationofthetoxicantisneeded .Traditio…     

Norditerpenoid and dianthramide glucoside alkaloids from cultivated<Emphasis Type="Italic">aconitum</Emphasis> species from Korea

A new norditerpene alkaloid named 8-O-methylhypaconine (1) was isolated along with twelve known alkaloids from the underground parts of an unknown species of Aconitum plant culti vated in Korea. Among the known alkaloids, two dianthramide glucosides, N-(2'-beta-glucopyra nosyl-5'-hydroxysalicyl)-5-hydroxyanthranilic acid methyl ester (2) and N-(2'-beta-glucopyranosyl-5'-hydroxysalicyl)-5-hydroxy-6-methoxyanthranilic acid methyl ester (3), were isolated from Aconitum plants for the first time. The structures were established on the basis of chemical and spectroscopic methods.     

Studies on structural elucidation of Aconitum diterpenoid alkaloid by LC-APCI-MS and effects of Aconitum diterpenoid alkaloid on cutaneous blood flow

The chemical constituents of Aconitum yesoense var. macroyesoense and Aconitum japonicum were examined using high-resolution spectral analysis. Twelve novel alkaloids were isolated from A. yesoense var. macroyesoense together with 20 known alkaloids. Eight novel alkaloids were isolated from A. japonicum together with 15 known alkaloids. An HPLC-atmospheric pressure chemical ionization-mass spectrometry (HPLC-APCI-MS) method was useful for the simultaneous determination of 21 Aconitum alkaloids found in A. yesoense var. macroyesoense and A. japonicum. These compounds were fairly stable under the conditions used, and the protonated molecules or fragment ions characteristic of the molecule appeared as base peaks in the mass spectra and were used for selected ion monitoring. HPLC-APCI-MS is a very promising approach for structural investigations of positional isomers and stereoisomers. This method was applied successfully to stereoisomeric Aconitum alkaloids differing in configuration at C-1, -6, or -12. Comparison of the APCI spectra showed that the abundance of fragment ions was significantly higher for the C-1, -6, or -12 beta-form alkaloid than for C-1, -6, or -12 alpha-form alkaloid. The main alkaloid constituents in the root of A. yesoense var. macroyesoense, Aconitum alkaloids of the C20-diterpenoid type, kobusine and pseudokobusine, and their acyl derivatives were examined for their peripheral vasoactivities by measuring laser-flowmetrically the cutaneous blood flow in the hind foot of mice after intravenous administration. It is thought that the hydroxyl groups of alkaloids, especially a free OH group of pseudokobusine at C-6, were important for action on the peripheral vasculature leading to dilatation, and the results indicated that esterification of the hydroxyl group at C-15 with either anisoate, veratroate, or p-nitroben-zoate may contribute to enhancement of the activity of the parent alkaloids.     

Determination of Aconitum alkaloids in the tubers of Aconitum japonicum using gas chromatography/selected ion monitoring

A rapid, specific and precise method using GC/SIM was applied to separate and quantify Aconitum alkaloids in Aconitum tubers. The TMS derivatives of each alkaloid produced a well defined peak on selected ion recording (SIR). Good linear response over the range of 100 pg - 7.5 ng was demonstrated for each alkaloid. Furthermore, we investigated the changes in the contents of Aconitum alkaloids, i.e., hypaconitine, mesaconitine, aconitine, and jesaconitine, in Aconitum tubers paying special attention to the harvesting season. Mesaconitine had the highest value in all seasons. Mesaconitine and aconitine showed the highest value in the samples harvested in May 1991 while hypaconitine had the highest value among those harvested in December 1990. As to the total amount of the alkaloids, the highest value, 4190 microg/g, and the lowest value, 1881 microg/g, were observed in the samples harvested in May and September, respectively. GC/SIM was very useful for the determination of Aconitum alkaloids in the tubers.     

乌头属药材特征成份二萜生物碱的电喷雾质谱研究进展

本文从乌头属药材中二萜生物碱类成分的鉴定、炮制过程对二萜生物碱类成分种类及其含量的影响、复方制剂中二萜生物碱的鉴定、二萜生物碱类成份的体外生物转化以及二萜生物碱的生物体内代谢过程研究等方面综述了近年来电喷雾质谱在以上研究中的应用。     

Diterpenoid alkaloids from the aerial parts of Aconitum anthora L

Isolation and identification of six diterpenoid alkaloids, from Aconitum anthora L. are described. All proton detected 2D NMR techniques have been used for unambiguous 1H and 13C chemical shift assignments of guan-fu-base-Y (6). This is the first investigation of the diterpenoid alkaloids of this plant species.     

草乌与贝母配伍禁忌的化学研究

目的:探讨草乌与贝母在煎煮过程及体外模拟消化环境中生物碱类成分的变化规律,初步阐明草乌配伍贝母毒性作用变化原因。方法:采用酸性染料比色法、改良异羟肟酸铁分光光度法和HPLC法,测定草乌与贝母不同配比合煎液及人工胃、肠液处理后溶液中总生物碱和酯型生物碱、乌头碱类成分的含量,并通过主成分分析(PCA)和聚类分析(HCA)综合考察各配比和处理方法对毒性作用变化情况。结果:通过PCA和HCA的综合考察,两者得到的结果相似,均显示11个样品被分为3组,其中样品1为Ⅰ组,样品2、5、9、10、11为Ⅱ组,样品3、4、6、7、8为Ⅲ组,样品的分类结果可能与草乌生物碱和贝母生物碱含量有关;草乌与贝母在一定的比例下毒性增加,尤其是其合煎剂经胃液处理后毒性进一步增大,而经过肠液处理后毒性有一定的降低。结论:"草乌反贝母"是在一定条件下的配伍禁忌,可能与配伍比例和药物体内释放部位有一定的关系。     

高乌头炮制品的HPLC指纹图谱研究及其2种生物碱成分含量测定

目的 建立不同产区高乌头炮制品的HPLC指纹图谱,并测定2种生物碱成分的含量,为高乌头制品质量控制提供参考依据。方法 通过HPLC梯度洗脱建立高乌头炮制品的指纹图谱,并采用中药指纹图谱相似度评价系统(2012版)、主成分分析(PCA)和聚类分析(CA)对指纹图谱进行分析。结果 建立了高乌头炮制品指纹图谱,10批高乌头制品的相似度均大于0.90,标定共有峰18个,并对其中2主要成分(高乌甲素、冉乌头碱)进行含量测定;聚类分析(CA)将所有批次高乌头制品分为4类,反映了10批不同产区高乌头制品的质量特征;主成分分析(PCA)筛选出累计贡献率达到88.824%的4个主成分,得到决定高乌头药材质量7个化学成分。结论 该方法重现性好、特征性强可用于高乌头炮制品全面质量评价。     

Distribution of Aconitum alkaloids in body fluids and tissues in a suicidal case of aconite ingestion

A case involving a suicidal ingestion of Aconitum tubers is presented. A 40-year-old woman in Hokkaido, Japan ingested ground aconite and died of aconite intoxication about 4 h after ingestion. The Aconitum alkaloids were quantitated using gas chromatography-selected ion monitoring from extracts of the body fluids and organs. The blood and urine concentrations of jesaconitine, the main alkaloid of the aconite in this case, were 69.1 ng/mL and 237.8 ng/mL, respectively. Higher values of the alkaloid were demonstrated in the kidneys, the liver, and in the bile rather than other organs or serum, suggesting the alkaloids were eliminated by the liver and kidneys. In the gastrointestinal tract, the highest value of jesaconitine (471.3 ng/g) was in the ileal contents. These findings show that Aconitum alkaloids were found in the liver and kidneys in much higher concentrations than in serum and suggest that they were eliminated not only via urine but also in feces. Feces may be useful to detect Aconitum alkaloid if other biological samples are not available.     

Development and validation of a highly sensitive UPLC-MS/MS method for simultaneous determination of aconitine, mesaconitine, hypaconitine, and five of their metabolites in rat blood and its application to a pharmacokinetics study of aconitine, mesaconitine, and hypaconitine

A rapid, specific and sensitive method was developed for the simultaneous determination of eight Aconitum alkaloids: aconitine (AC), mesaconitine (MA), hypaconitine (HA), benzoylaconine (BAC), benzoylmesaconine (BMA), benzoylhypaconine (BHA), aconine and mesaconine in rat blood by ultra-performance liquid chromatography coupled tandem mass spectrometry (UPLC-MS/MS). The UPLC-MS/MS system coupled with an electrospray ionization (ESI) source was operated in a positive mode via multiple-reaction monitoring (MRM). Samples were treated with methanol to remove protein prior to analysis by UPLC-MS/MS. The analytes were separated with a Waters C18 column (1.7 μm, 50?×?2.1?mm) and a gradient elution using acetonitrile and 0.1% formic acid-water as the mobile phases. The linear response range was from 0.125 to 1000 nmol/L for these eight alkaloids and the correlation coefficients (r(2) values) were all higher than 0.997. The method was validated with respect to precision, accuracy, recovery, matrix effect, carryover effect and sample stability, and found to be within the acceptable limits. The developed and validated method was successfully applied to simultaneously determine the eight Aconitum alkaloids in rats blood after intravenous administration of a mixture of AC, MA and HA.     

直缘乌头根中生物碱成分的研究

目的：研究直缘乌头（Aconitum transsectum Diels.）根中的生物碱成分。方法：采用pH梯度、中压柱层析、常压硅胶柱层析及离心薄层层析法。结果：分得11个已知二萜生物碱，由光谱法分别鉴定为：滇乌碱（yunaconitine）、印乌碱（indaconitine）、黄草乌碱甲（vilmorrianine A）、黄草乌碱丙（vilmorrianine C）、草乌甲素(crassialine A)、膝乌碱（geniconitine）、查斯曼宁（chasmanine）、８－去乙酰滇乌碱（8-deacetylyunaconitine）、塔拉萨敏（talatisamine）、14-乙酰塔拉萨敏（14-acetyltalatisamine）和chasmaconitine。结论：黄草乌碱甲和chasmaconitine是首次从该植物中分离得到。     

不同品种附子生物碱和多糖含量的比较

目的:比较不同品种附子生物碱和多糖的含量。方法:采用酸性染料比色法测定附子总生物碱含量,反相高效液相色谱法测定双酯型生物碱含量,蒽酮-硫酸法测定多糖含量。结果:不同品种附子总生物碱和双酯型生物碱含量差异较大,附子多糖含量差异不显著。附子炮制后,黑附片中总生物碱、双酯型生物碱含量显著下降,附子多糖含量稍有增加。结论:本试验结果可为附子品种选育提供一定的科学依据。     

露蕊乌头生物碱的研究(Ⅰ)

从藏药露蕊乌头(Aconitum gymnandrum Maxim)中分得四个生物碱,经化学和光谱鉴定,其中两个为已知生物碱atisine.Hcl(A)和talatisamine(B),另两个为新的二萜类生物碱,分别命名为露乌碱,(C)(gymnaconitine)和甲基露乌碱(D)(mgthyl gymnaconitine),这两种新的生物碱均联接有二甲基咖啡酸酯:在乌头生物碱中是首次发现。     

黄花乌头中Hetisine型生物碱的高速逆流色谱分离与结构鉴定

目的研究黄花乌头块根的化学成分，寻找更多天然活性物质。方法采用高速逆流色谱法分离纯化黄花乌头块根中的生物碱类化学成分，根据理化性质、波谱学分析鉴定化合物的结构。结果高速逆流色谱的两相溶剂分离系统采用氯仿-甲醇-0.2 mol·L^-1 HCl(体积比为10∶3∶3)，从黄花乌头块根中一次性分离得到8个化合物，分别鉴定为2α-propionyl-11α,13β-diacetyl-14-hydroxyhetisine (I)、关附巳素(II)、关附庚素(III)、关附己素(IV)、关附Z素(V)、关附辰素(VI)、关附甲素(VII)、关附壬素(VIII)。结论化合物I为新化合物，命名为关附未素(Guanfu base R)。     

太白乌头中一新的去甲二萜生物碱

为了研究太白乌头(Aconitum taipaicum)根的化学成分，采用硅胶柱色谱进行分离纯化，根据其理化性质和波谱数据确定化合物结构。从该植物的根中分离得到1个新的去甲二萜生物碱isodelelatine (1)和5个已知生物碱，分别为atisine (2)、 delfissinol (3)、 liangshanine (4)、 hypaconitine (5)和delelatine (6)。化合物2～6均为首次从该植物中分离得到。