Isolation and chemistry of alkaloids from plants of the family Papaveraceae LXVII: Corydalis cava (L.) Sch. et K. (C. tuberosa DC).

From Corydalis cava (L.) Sch. et K. (C. tuberosa DC) (Papaveraceae: genus Corydalis Med.), a mixture of alkaloids (0.53%) was isolated. The main alkaloid was (+)-bulbocapnine, and the minor alkaloids were coptisine, (+)-domestine, adlumidiceine, (+)-predicentrine, protopine, (-)-capnoidine, (+)-stylopine, (+)-isoboldine, 8-oxocoptisine, 1,2-methylenedioxy-6a,7-dehydroaporphine-10,11-quinone, and corysamine. In addition, fumaric acid was obtained. Coptisine, adlumidiceine, predicentrine, 8-oxocoptisine, corysamine, 1,2-methylenedioxy-6a, 7-dehydroaporphine-10,11-quinone, and isoboldine were found in C. cava for the first time, but rhoeadine and papaverrubine alkaloids were not detected. Predicentrine and isoboldine were identified on the basis of the UV, IR, mass, and PMR spectra.     

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.     

基于HPLC-Q-TOF/MS的白屈菜生物碱化学组成快速辨识研究

利用HPLC-Q-TOF/MS技术快速准确、系统全面的定性鉴定白屈菜中生物碱。采用XCharge C18色谱柱(5μm, 4.6 mm×250 mm);以乙腈-0.1%甲酸为流动相梯度洗脱;流速0.7 mL·min-1。根据一级质谱分子离子和二级质谱碎片离子,归纳白屈菜生物碱的质谱裂解规律,结合相关文献,对白屈菜中生物碱进行系统的结构鉴定。为验证结果的准确性,纯化制备其中3个生物碱并通过核磁技术鉴定化学结构。从白屈菜中鉴定出21个生物碱,包含1个阿朴菲类生物碱、3个普罗托品类生物碱、11个苯并菲啶类生物碱以及6个原小檗碱类生物碱。其中N-甲基金罂粟碱(8)为白屈菜中首次报道,二氢黄连碱(11)和去甲基白屈菜碱(12)为白屈菜中首次用该技术鉴定。制备化合物的化学结构鉴定结果与质谱定性结果一致。经验证,该方法快速准确,可为白屈菜化学成分的分析鉴定和提取分离提供依据。     

Qualitative and quantitative determination of ten alkaloids in traditional Chinese medicine Corydalis yanhusuo W.T. Wang by LC-MS/MS and LC-DAD

An analytical method incorporating high-performance liquid chromatography (HPLC) with MS and UV-detection was developed for the qualitative and quantitative determination of alkaloids in Corydalis yanhusuo. Ten alkaloids, including seven tertiary alkaloids and three quaternary alkaloids, were identified by comparing their retention times, UV and MS spectra with those of authentic compounds. Furthermore, the collision-induced dissociations of the [M+H](+) and [M](+) ions were studied to clarify the MS behavior of the different types of alkaloids. In positive ion electrospray ionization tandem mass spectrometry (ESI-MS) all the tertiary alkaloids yielded prominent [M+H](+) ions and quaternary alkaloids yielded prominent [M](+) ions in the first order mass spectra. Fragments involving losses of H, CH(3), CO, H(2)O and OCH(3) were observed in the MS/MS spectra. In addition, quantification of the 10 alkaloids in Corydalis yanhusuo from methanol and ethyl acetate extract of different origins were performed by this method, which provides a new tool for the assessment of quality of Corydalis yanhusuo preparations. The method provides the best sensitivity and specificity for characterization and quantitative determination of the alkaloids in Corydalis yanhusuo so far.     

Isolation and identification of three new flavones from Achillea millefolium L.

Column chromatography on silica gel of a petroleum ether extract of the flowering heads of Achillea millefolium L. allowed three flavones to be separated and identified. Spectral studies (PMR, mass spectrometry, and UV) and a comparison with data for compounds reported in the literature established the flavones as 5-hydroxy-3,6,7,4'-tetramethoxyflavone, artemetin, and casticin. These compounds have not been reported previously as constituents of A. millefolium.     

Alkaloids and flavonoid from aerial parts of Hammada articulata ssp. scoparia

Two alkaloids (N-methylisosalsoline and carnegine) had been previously described from the aerial parts of Hammada articulata ssp. scoparia. A thorough study of this plant material has now led to the isolation of eight minor alkaloids and one flavonoid. The alkaloids include four isoquinolines (isosalsoline, salsolidine, dehydrosalsolidine and isosalsolidine), one isoquinolone (N-methylcorydaldine), tryptamine, N-omega-methyltryptamine and one beta-carboline (tetrahydroharman). The flavonoid has been identified as isorhamnetin-3-O-beta-D-robinobioside. The structures have been elucidated on the basis of spectral data, mainly mass spectrometry (D-IC) and 1H NMR.     

Alkaloids of Haloxylon salicornicum (Moq.) Bunge ex Boiss. (Chenopodiaceae)

Haloxylon salicornicum is a desert plant that contains several alkaloids. From the aerial parts a new piperidyl alkaloid, haloxynine, was isolated and characterized on the basis of mass spectrometry, 1H and 13C NMR. A GLC/MS analysis revealed the presence of 17 additional known alkaloids of which piperidine, halosaline, anabasine, hordenine, N-methyltyramine, haloxine and aldotripiperideine had been previously reported in this genus. Among the 18 identified alkaloids, ten alkaloids were recorded for the first time from this plant and the genus Haloxylon. Haloxynine, halosaline, haloxine, anabasine, and smipine figure as major alkaloids with a relative abundance of more than 5% of total alkaloids. Some of these alkaloids are known be strong agonists at nicotinic acetylcholine receptors and it is thus likely that they serve as chemical defencecompounds against insects and mammalian herbivores.     

Synthesis and antimicrobial activity of n,7-diaryl-5-methyl-4,7-dihydrotetrazolo[1,5-<Emphasis Type="Italic">a</Emphasis>] pyrimidine-6-arboxamides

N,7-Diaryl-5-methyl-4,7-dihydrotetrazolo[1,5-a]pyrimidine-6-carboxamides were synthesized by three-component reaction of acetoacetanilides (2-methylacetoacetanilide, 2,4-dimethylacetoacetanilide, 4-chloroacetoacetanilide, o-acetoacetanilide) with a mixture of aromatic aldehyde and 5-aminotetrazole. The proposed structures are confirmed by IR and PMR spectroscopy and mass spectrometry. The synthesized compounds are characterized with respect to antimicrobial activity.     

Detecting Taxus poisoning in horses using liquid chromatography/mass spectrometry

A method is described for the analysis of taxine alkaloids by liquid chromatography/mass spectrometry. It is applicable to the detection of taxine alkaloids in the stomach contents of horses in which Taxus poisoning is suspected. Analysis of a leaf extract of Taxus baccata revealed unreported alkaloids of the same relative molecular mass as taxine B and isotaxine B.     

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。     

Metabolite fingerprinting of Camptotheca acuminata and the HPLC–ESI-MS/MS analysis of camptothecin and related alkaloids

The major phytochemical constituents, namely, alkaloids, flavonoids and ellagic acid derivatives, of leaves of Camptotheca acuminata were identified using high performance liquid chromatography (HPLC) coupled with electrospray mass spectrometry (ESI-MS) in extracts of plants cultivated in Italy and collected at different growth stages. Alkaloids related to camptothecin were identified and quantified by HPLC coupled with ESI-tandem mass spectrometry (MS/MS) employing, respectively, an ion trap and a triple quadrupole mass analyser. The fragmentation patterns of alkaloids related to camptothecin were analysed and a specific Multiple Reaction Monitoring HPLC–MS/MS method was developed for the quantitative determination of these constituents. The described method provides high sensitivity and specificity for the characterisation and quantitative determination of the alkaloids in C. acuminata.     

Rapid and global detection and characterization of aconitum alkaloids in Yin Chen Si Ni Tang,a traditional Chinese medical formula,by ultra performance liquid chromatography–high resolution mass spectrometry and automated data analysis

An improved method employing Metabolynx XS with mass defect filter (MDF), a post-acquisition data processing software, was developed and applied for global detection of aconitum alkaloids in Yin Chen Si Ni Tang, a traditional Chinese medical formula (TCMF). The full-scan LC–MS/MS data sets with extra mass were acquired using ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS) with the MS^E mode in a single injection. To remove the interferences, Metabolynx XS was optimized to extract the ions of aconitum alkaloids located at the lower abundance. As a result, 62 ions were assigned rapidly to aconitum alkaloids and identified tentatively by comparing the accurate mass and fragments information with that of the authentic standards or by mass spectrometry analysis and retrieving the reference literatures. Compared with the previous studies on Fuzi-containing TCMF, the report detected more aconitum alkaloids, and the analysis process was accelerated by automated data processing. It is concluded that the screening capability of Metabolynx XS with MDF, together with the utilization of MS^E in structural elucidation, can facilitate a rapid and comprehensive searching and effective structural characterization of aconitum alkaloids in TCMF.     

液相色谱-质谱法测定雷公藤浸膏中4种倍半萜类生物碱含量

目的建立一种灵敏、可靠的雷公藤浸膏中4种倍半萜类雷公藤生物碱含量的高效液相色谱-质谱检测方法。方法雷公藤浸膏样品经氯仿溶解后用稀盐酸（2.0 mol·L~（-1））提取,Waters Oasis.MCX小柱进行净化,以0.05%（V/V）醋酸-醋酸铵溶液（5 mmol·L~（-1））/乙腈（45/55,V/V）为流动相,采用Zorbax SB C_（18）柱（250mm×4.6mm,5μm）分离,应用高效液相色谱/大气压化学电离正离子选择离子监测模式（SIM）测定,雷公藤春碱、雷公藤定碱、雷公藤吉碱和雷公藤次碱的定量离子分别为m/z874、884、858和868。结果 4种雷公藤生物碱的绝对回收率为86.5%～96.0%,质量浓度在1.0～200.0μg·L~（-1）内具有良好线性,批内RSD〈7.4%,批间RSD〈9.3%,定量检出限均为1.0μg·kg~（-1）。结论本方法简便、灵敏、干扰少、特异性好,可用于雷公藤浸膏中4种倍半萜类雷公藤生物碱含量的检测。     

基于UPLC-Q-TOF-MS技术对黄连须生物碱类成分及其裂解规律的分析

目的 应用超高效液相色谱-飞行时间质谱联用技术（ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry,UPLC-Q-TOF-MS）分析黄连须中生物碱类成分,并对其中主要类型生物碱的裂解规律进行解析。方法 采用Waters Acquity UPLC BEH C₁₈色谱柱,以0.1%甲酸水（A）-0.1%甲酸乙腈（B）为流动相,梯度洗脱,质谱采用电喷雾离子源,正负离子模式下采集数据,在Peakview 2.0/masterview 1.0软件中依据自建的黄连化学成分数据库筛查化合物,通过精确质量数及比对Natural Products HR-MS/MS Spectral Library 1.0 software所含的1 000余种中草药对照品二级谱图进行裂解规律分析。结果 鉴定并推断出黄连须中19个生物碱类成分,并对其中原小檗碱型、氧化小檗碱型及普罗托品型生物碱的裂解规律进行了分析。结论 该法为黄连须成分研究提供了依据,证明了黄连须的利用价值,并对其进一步开发利用奠定了基础。     

Variability of alkaloids in the skin secretion of the European fire salamander (Salamandra salamadra terrestris).

The two major alkaloids, samandarine and samandarone, were identified in the skin secretion of individual specimens from two populations of the European fire salamander (Salamandra salamandra terrestris) by gas chromatography/mass spectrometry. High intraspecific variability in the ratio of both alkaloids was observed, but also in individual specimens over a period of 4 months suggesting separate metabolic pathways of the compounds. Alkaloid synthesis appears to take place also in liver, testes and ovaries, whereas the larvae of the salamanders are entirely free of alkaloids.     

Pyrrolizidine alkaloids: Organohalogen derivative isolated from Senecio jacobaea (Tansy Ragwort)

Senecio jacobaea (Tansy Ragwort) has previously been reported to contain 6 hepatotoxic pyrrolizidine alkaloids (PAs). Utilizing high pressure liquid chromatography (HPLC) plus mass spectrometry, 3 additional pyrrolizidine alkaloids are described, including a second chlorinated pyrrolizidine alkaloid. These additional pyrrolizidine alkaloids, through a variety of products, appear to pose an increased human health hazard.     

Characterization of Sanguinaria canadensis L. fluid extract by FAB mass spectrometry.

Positive-ion fast atom bombardment mass spectrometry was used for the rapid characterization of commercial Sanguinaria canadensis L. fluid extracts. Quaternary and non-quaternary benzophenanthridine alkaloids afford persistent peaks due to [M]+ and [M+H]+ ionic species, respectively, and their relative abundances are in good agreement with previously reported per cent analytical data. The procedure allowed sanguinarine, chelerythrine, chelirubine, sanguilutine, protopine, allocryptopine and the isomers sanguirubine and/or chelilutine to be effectively detected by means of persistent and intense peaks in all the samples examined.     

Structural analyses of protoberberine alkaloids in medicine herbs by using ESI-FT-ICR-MS and HPLC-ESI-MS(n)

The Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FT-ICR-MS) using sustained off-resonance irradiation (SORI)/collision-induced dissociation (CID) method at high mass resolution has been first applied to investigate the characteristic fragment ions of four protoberberine alkaloids in medicine herbs. The ESI-FT-ICR SORI-CID experiment results demonstrate that the unambiguous elemental composition of fragment ions can be obtained at high mass resolution, then the logical fragmentation pathways of the protoberberine alkaloids has been proposed. The characteristic fragment ions of CID and MS(n) of protoberberine alkaloids have been discussed, which are specific and useful for the identification of some protoberberine alkaloid compounds. Then, the extracts of four kinds of medicine herbs have been analyzed by HPLC-ESI-MS(n). According to these characteristic fragmentation pathways, the retention time (t(R)) of HPLC and mass spectra of product ion, the structures of six kinds of protoberberine alkaloids have been identified. And, in the present paper, the selected ion monitoring (SIM) method has been used to separate and identify the alkaloid isomers.     

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。     

Characterization of alkaloids in Sophora flavescens Ait. by high-performance liquid chromatography-electrospray ionization tandem mass spectrometry

Sophora flavescens Ait., a well-known Chinese herbal medicine, is widely used in clinical practice for the treatment of viral hepatitis, cancer, gastrointestinal hemorrhage, and skin diseases. This paper is the first report on a method based on the combined use of high-performance liquid chromatography, photodiode array detection, and electrospray ionization tandem mass spectrometry for the comprehensive and systematic separation and characterization of bioactive alkaloids in Sophora flavescens Ait. A total of 22 constituents were identified on the basis of the extracted ion chromatograms for different [M+H](+) ions of the alkaloids present in S. flavescens Ait. Among these, 5 constituents were unambiguously identified by comparing the experimental data on their retention times and MS(n) spectra with those of the authentic compounds, and 17 other constituents were tentatively identified on the basis of their MS(n) fragmentation behaviors and/or molecular weight information from literatures. Furthermore, some characteristic fragmentation pathways of the alkaloids in S. flavescens Ait. were detected and examined. This information may be useful for characterizing the bioactive alkaloids present in S. flavescens Ait. and for possible applications in formulations.