Studies of the intestinal absorption of the alkaloids in the Wu-tou decoction combined with different incompatible medicinal herbs in a Caco-2 cell culture system using UPLC-MS/MS

In this study, seven alkaloids were detected in Wu-tou decoction using ultra performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MSn). The aim of this study was to investigate the effect of Fritillariae Cirrhosae Bulbus, Fritillariae Thunbergii Bulbus, Pinelliae Rhizoma in different ratios with Wu-tou decoction (2:1, 1:1, 1:2) by measuring the therapeutic effects in Wu-tou decoction of main seven alkaloids including benzoylaconitine (BA), benzoylmesaconitine (BM), benzoylhypaconitine (BH), hypaconitine (HA), fuziline (FU), niaolin (NE) and deoxyaconitine (DA). The permeability of aconitum alkaloids extract through a Caco-2 cell monolayer was analyzed in the absence and presence of Fritillariae Cirrhosae Bulbus, Fritillariae Thunbergii Bulbus, and Pinelliae Rhizoma, respectively. The results showed that Pinelliae Rhizoma could reduce the absorption of the alkaloids and increase the excretion of the alkaloids, which would attenuate the therapeutic effects of Wu-tou decoction. Therefore, Pinelliae Rhizoma is an incompatible herb of Wu-tou decoction because of the inhibition of the absorption of alkaloids in the intestine. And that Fritillariae Cirrhosae Bulbus and Fritillariae Thunbergii Bulbus showed the effects to improve the permeability of the alkaloids in Wu-tou decoction. These effects of these two herbs were similar, but the former was stronger than the latter, which most likely is due to the fact that the compositions of these two traditional Chinese medicines are similar. The in vitro data suggests that the compounds such as fritillary presented in alkaloids in the formula maybe improve the therapeutic function caused by the increased bioavailability of alkaloids in intestine.     

利用液质联用技术分析干姜对乌头类生物碱在大鼠肠内吸收的影响

为从吸收的角度考察干姜对乌头类双酯型生物碱的解毒机理, 采用外翻肠囊法展开实验. 利用超高液相与三重四极杆质谱联用技术定量检测双酯型生物碱成分, 采用标准曲线法计算乌头碱、中乌头碱、次乌头碱在肠囊内吸收的绝对含量, 采用质谱峰面积直接分析其它双酯型生物碱的相对变化, 结果加入干姜提取液后, 乌头碱、中乌头碱、次乌头碱的单位肠管面积累计吸收量均降低, 10-羟基中乌头碱的的累积峰面积降低; 加入维拉帕米后, 双酯型生物碱的单位肠管面积累计吸收量及累积峰面积均增加; 向含有地高辛的肠营养液中加入干姜提取液后, 地高辛在各实验时间点的单位肠管面积累计吸收量均降低, 根据以上结果推测干姜抑制乌头类双酯型生物碱在大鼠肠囊内吸收的可能机制是通过诱导肠内P-葡糖蛋白, 从而抑制作为P-葡糖蛋白底物的双酯型生物碱的吸收, 最终起到减毒作用.     

生附子中C19型二萜生物碱的高速逆流色谱分离及其结构鉴定

应用高速逆流色谱法分离制备了生附子中的3个C19型二萜生物碱类化合物。以正己烷-乙酸乙酯-甲醇-水(3:5:4:5, v/v/v/v)为两相溶剂系统,上相为固定相,下相为流动相,在主机转速850 r/min、流动相流速2.0 mL/min、检测波长235 nm条件下进行分离制备;一次性从90 mg附子总碱粗提物中分离制备得到15.3 mg北草乌碱,35.1 mg中乌头碱和22.7 mg次乌头碱,经高效液相色谱分析,测得它们的纯度分别为97.9%、96.2%和99.2%。并应用波谱(电喷雾离子质谱、核磁共振氢谱和核磁共振13C谱)解析法确定了它们的结构。利用该方法可以对生附子中的二萜类生物碱成分进行快速的分离和纯化     

Analysis of Norditerpenoid Alkaloids. Extracted from Aconitum sinomantanum Nakai by Electrospray Ionization Tandem Mass Spectrometry

Introduction AconitumsinomantanumNakai(GaowutouinChi nese)isdistributedinthenorthwesternareaofChina,anditsroots(RAS)areexternallyusedasakindof folkmedicineinChina.Themainactiveconstituentsof RASarenorditerpenoidalkaloids.Themajoralkaloid,lappaconitine,was…     

Norditerpenoid alkaloids from the processed tubers of Aconitum carmichaeli

Four new and five known norditerpenoid alkaloids were isolated from the processed tubers of Aconitum carmichaeli. The new alkaloids are 14-O-cinnamoylneoline (3), 14-O-anisoylneoline (4) 14-O-veratroylneoline (5), and lipo-14-O-anisoylbikhaconine (8). The known alkaloids are neoline (1), 14-O-acetylneoline (2), foresaconitine (6), crassicauline A (7), and lipohypaconitine (9). Alkaloids 2, 6, and 7 were isolated from this plant for the first time. The structures of the new alkaloids were established by spectroscopic and chemical methods.     

Unprecedented C_(19)-diterpenoid alkaloid glycosides from an aqueous extract of “fu zi”: Neoline 14-O-L-arabinosides with four isomeric L-anabinosyls

Four structurally unprecedented aconitane-type C_(19)-diterpenoid alkaloid glycosides with isomeric arabinosyls, named aconicarmichosides A–D(1–4), were isolated from an aqueous extract of "fu zi", the lateral roots of Aconitum carmichaelii. Their structures were determined as neoline 14-O-a-and 14-O-b-L-arabinopyranosides(1 and 2) and 14-O-a-and 14-O-b-L-arabinofuranosides(3 and 4), by spectroscopic and chemical methods including 2D NMR experiments and acid hydrolysis. Compounds 1–4 represent the first examples of glycosidic diterpenoid alkaloids.     

Structure identification of five unknown dilipo-alkaloids extracted from processed tuber of Aconitum Carmiechaeli by electrospray ionization tandem mass spectrometry.

The alkaloids in processed aconite tuber of Aconitum Carmiechaeli were studied, and five novel alkaloids in extract from processed aconite tuber were found. The first step involved the use of electrospray ionization mass spectrometry (ESI-MS), and then multi-stage tandem mass spectrometry (MSn) was used to provide structural information. Based on their MSn spectra, the structures of the five novel compounds were elucidated to be C3,C8-difatty acid esters of mesaconitine, aconitine and 10-hydroxyaconitine.     

成核剂对聚2,6-萘二甲酸乙二酯结晶行为的影响

通过DSC、偏光显微镜 (POM)和小角激光散射 (SALS)研究了苯甲酸钠 (SB)对聚 2 ,6 萘二甲酸乙二酯(PEN)结晶行为的影响 .研究发现SB是PEN的成核剂 .在高温下 ,SB与PEN发生化学反应 ,生成可以起到成核作用的物质 ,降低了PEN的成核表面自由能位垒 ,提高了PEN在高温区的结晶速率 .同时 ,SB还可以提高PEN在低温区的结晶速率 .此外 ,SB的加入还使PEN分别在高温区和低温区结晶得到的多角晶和球晶尺寸减小 ,这表明不管在高温区还是在低温区 ,SB都可以提高PEN的成核速率 .而含有SB的PEN结晶样品的SALS图像变得弥散、模糊 ,则说明SB使PEN在低温下结晶得到的球晶变得不完善     

草乌中二萜类生物碱的电喷雾串联质谱研究

利用电喷雾质谱技术对传统中药草乌中的二萜类生物碱进行直接分析鉴定.通过实验数据并对比文献发现,草乌中含有单酯型、双酯型、三酯型和脂类等共4种类型生物碱,其中三酯型和脂类生物碱在草乌中为首次发现.由于这些生物碱的结构相似,在电喷雾串联质谱中碎裂方式相同,因此根据电喷雾串联质谱结果确定了这些生物碱的结构.     

Qualitative and quantitative analysis of aconitine-type and lipo-alkaloids of Aconitum carmichaelii roots

By optimizing the extraction and analytical conditions, a reliable and precise HPLC method coupled with photodiode array detection (HPLC–DAD) has been developed for the identification and quantification of three major aconitine-type alkaloids (aconitine, mesaconitine, hypaconitine) in the roots of Aconitum carmichaelii Debeaux. The qualitative analysis of the plant material was carried out by LC-APCI-MSⁿ. By means of this method, 26 lipo-alkaloids were also identified from the roots of A. carmichaelii. The effect of processing on aconitine-type alkaloids, lipo-alkaloids and pure aconitine was studied. As part of our investigation, two lipo-alkaloids, 14-benzoylaconine-8-palmitate and 14-benzoylaconine-8-linoleate were produced semisynthetically. The COX-1, COX-2 and LTB₄ formation inhibitory activity of aconite root extracts and different types of diterpene alkaloids and the toxicity of lipo-alkaloids were also investigated.     

Hemsleyaconitines F and G,Two Novel C19‐Diterpenoid Alkaloids Possessing a Unique Skeleton from Aconitum hemsleyanum

Hemsleyaconitines F and G ( 1 and 2 , resp.) were isolated from the EtOH extract of Aconitum hemsleyanum. Their structures were elucidated by extensive analyses of the IR, 1D‐ and 2D‐NMR, and MS data. The two C₁₉‐diterpenoid alkaloids 1 and 2 possess a novel skeleton, featuring a five‐membered D‐ring between C(9), C(13), C(14), C(15), and C(16), which is quite different from the previously isolated six‐membered D‐ring analogs.     

HPLC测定植物性农药——0.25%乌头总碱乳油中的乌头类生物碱

 选择十八烷基键合相柱 ,以甲醇 水 氯仿 三乙胺 (体积比为 6 8∶32∶2∶0 1)混合溶液为流动相 ,用高效液相法测定了一种植物性农药 0 2 5 %乌头总碱乳油中的乌头生物碱。实验结果表明中乌头碱、乌头碱及次乌头碱与其他杂质能够得到很好的分离。以安宫黄体酮作内标物 ,用峰面积比测定各生物碱含量 ,在其线性范围内分析结果准确 ,回收率高 (>92 % ) ,重现性好 (RSD <3 2 % ) 。     

蒙药草乌花及其提取物化学成分的红外光谱法整体结构解析

采用傅立叶变换红外光谱法并借助于二阶导数谱以及二维相关红外光谱(2D-FTIR)研究了蒙药草乌花原药材及不同溶剂提取物(依次采用乙醚,乙醇和水提取)所含化学成分的红外谱图整体变化规律.结果显示:在草乌花原药材中具有明显的多糖类化合物的红外特征吸收峰,证明该药材中含有大量多糖类成分.草乌花药材乙醚和醇提物的红外光谱较相似...     

液相色谱/质谱/质谱联用法测定乌头属药材及中成药中乌头类生物硷含量

探讨了用LC／MS／MS联用法测定乌头属药材及有关中成药中乌头硷、次乌头硷和中乌头硷含量的方法。应用优化后的技术条件及采用多反应监测扫描方式,结果表明分析样品中各乌头硷浓度在0．5－40mg/L范围内,色谱峰面积与浓度有良好的线性关系。此法亦应用于14个不同的乌头属药材及10个不同的中成药样品加的乌头硷含量检测。     

直缘乌头中新的C19—二萜生物碱的结构研究

从直缘乌头（Aconitum transsectum Diels.）根中分离得到4个新的乌头碱型 C_(19)-二萜生物碱。由光谱法（IR, ~1H NMR, ~(13)C NMR, 2D-NMR, MS）确定其 结构为:8-O-乙基滇乌碱（8-O-ethylyunaconitine） (1)、去氮乙基查斯曼宁（N- deethyl chasmanine）(2)、直缘乌碱丁（transconitine D）(3)和直缘乌碱戊（ transconitine E）(4)。     

Puberunine and puberudine, two new C18-diterpenoid alkaloids from Aconitum barbatum var. puberulum

Two C(18)-diterpenoid alkaloids, puberunine (1) and puberudine (2), together with four other new alkaloids, including the first examples having β-oriented substitution at C-3 and a rare chloro-substituent were isolated from Aconitum barbatum var. puberulum. Their structures were elucidated by spectroscopic methods. Puberunine and puberudine, which possess a unique rearranged E ring and an opened A ring, respectively, represent new subtypes of the C(18)-diterpenoid alkaloids. A plausible biosynthetic pathway of 1 and 2 was proposed.     

Cardioactive C₁₉-diterpenoid alkaloids from the lateral roots of Aconitum carmichaeli "Fu Zi"

Bioassay-guided fractionation of an n-BuOH extract of the lateral roots of Aconitum carmichaeli. led to the isolation of 5 cardioactive C(19)-diterpenoid alkaloids: N-deethylaconine (1), beiwutinine (2), hypaconine (3), mesaconine (4), and 15α-hydroxyneoline (5). N-Deethylaconine and beiwutinine are new aconitine-type C(19)-diterpenoid alkaloids. Hypaconine was isolated from this species for the first time. Among them, mesaconine, hypaconine, and beiwutinine showed the strongest cardiac actions on the isolated perfused bullfrog heart. Furthermore, mesaconine has protective effects, including improved inotropic effect and left ventricular diastolic function, on myocardial ischemia-reperfusion injury in rat at a dose of 10(-9) mol/L. However, mesaconine has almost no effect on heart rate.     

Facile separation and determination of Aconitine alkaloids in traditional Chinese medicines by CE with tris(2,2'-bipyridyl) ruthenium(II)-based electrochemiluminescence detection

A facile CE method coupled with tris(2,2'-bipyridyl) ruthenium(II)-based electrochemiluminescence [Ru(bpy)(3) (2+)] detection was developed for simultaneous determination of Aconitum alkaloids, i.e., hypaconitine (HA), aconitine (AC), and mesaconitine (MA) in baseline separation. The optimal separation of these Aconitum alkaloids was achieved in a fused-silica capillary column (50 cm x 25 microm id) with 30 mM phosphate solution (pH 8.40) as running buffer at 12 kV applied voltage. The three alkaloids can be determined within 10 min by a single run. The calibration curves showed a linear range from 2.0 x 10(-7) to 2.0 x 10(-5) M for HA, 3.4 x 10(-7) to 1.7 x 10(-5) M for AC, and 3.8 x 10(-7) to 1.9 x 10(-5) M for MA. The RSDs for all analytes were below 3.01%. Good linear relationships were found with correlation coefficients for all analytes exceeding 0.993. The detection limits were 2.0 x 10(-8) M for HA, 1.7 x 10(-7) M for AC, and 1.9 x 10(-7) M for MA under optimal conditions. This method was successfully applied to determine the three alkaloids in Aconitum plants.     

Structural characterization and identification of C19‐ and C20‐diterpenoid alkaloids in roots of Aconitum carmichaeli by rapid‐resolution liquid chromatography coupled with time‐of‐flight mass spectrometry

Aconite alkaloids from the roots of Aconitum carmichaeli (Fuzi, in Chinese) have been investigated by rapid‐resolution liquid chromatography coupled with time‐of‐flight mass spectrometry (TOFMS) in positive mode. With dynamic adjustment of the key role as fragmentor voltage in TOFMS, an efficient transmission of the ions was achieved to obtain the best sensitivity for providing the molecular formula for each analyte, and abundant fragment ions for structural information. Fifteen authentic standards isolated from Fuzi with various structures were first characterized by TOFMS, including diester‐diterpenoid alkaloids (DDAs), monoester‐diterpenoid alkaloids (MDAs), alkylol amine‐diterpenoid alkaloids (ADAs), veatchine‐type alkaloids and atisine‐type alkaloids. Fragmentation rules and key diagnostic fragment ions have been summarized, and possible pathways of fragmentation have been proposed. By accurate mass measurements within 5 ppm error for each ion, 30 C₁₉‐diterpenoid alkaloids including 10 DDAs, 3 MDAs, 9 ADAs and 8 other type alkaloids, and 8 C₂₀‐diterpenoid alkaloids including 4 veatchine‐type alkaloids and 4 atisine‐type alkaloids could be identified in a methanolic extract of Fuzi. Some isomers of aconite alkaloids were also differentiated. Based on the differences between their fragmentation pathways and special fragment ions, each type of aconite alkaloids was differentiated. Copyright © 2009 John Wiley & Sons, Ltd.