配伍前后四逆汤中酯型生物碱的变化规律

目的 研究四逆汤中附子配伍不同药味前后,汤液中酯型生物碱的变化规律.方法 采用HPLC法测定附子单煎液、附子干姜合煎液、附子甘草合煎液和四逆汤中单酯型生物碱及双酯型生物碱成分的含量.结果 附子配伍不同药味之后,苯甲酰乌头原碱等3种单酯型生物碱的含量均升高,依次为四逆汤＞附子甘草合煎液＞附子干姜合煎液＞附子单煎液;仅在附子单煎液中检出次乌头碱,其浓度为0.11 mg·g-1,而在附子干姜合煎液、附子甘草合煎液、四逆汤中均未检测到双酯型生物碱成分.结论 附子在配伍其他药味后,单酯型生物碱成分的含量升高、双酯型生物碱成分的含量减低,为“附子无干姜不热、干姜缓附子之毒”与附子-甘草“相畏/相杀”减毒配伍“毒减而效不减”提供了依据;四逆汤组单酯型生物碱成分的含量上升幅度最大,体现了全方药简力专、大辛大热,使阳复厥回通,共奏回阳救逆之功.     

附子不同配伍药对中生物碱成分的电喷雾质谱分析

以传统中医用药附子的配伍药对为研究对象，考察附子与不同中药配伍对附子中生物碱的影响规律，揭示配伍减毒的科学内涵。利用电喷雾质谱技术和内标法，分别对生附子，生附子加炙甘草、白芍、干姜、大黄共煎液和药渣中乌头类生物碱进行系统考察。与生附子相比，双酯型生物碱在附子加炙甘草、大黄、白芍、干姜共煎液中的含量降低；附子加炙甘草、白芍、干姜共煎液中的脂型生物碱含量增高。附子与炙甘草、白芍、干姜配伍的解毒机制是使毒性较大的双酯型生物碱转化为毒性小的脂型生物碱；与大黄配伍的解毒机制是药物所含成分与附子中的双酯型生物碱络合生成难溶于水的复合物，使双酯型生物碱的含量降低。本法对深入研究乌头属植物的配伍作用机制具有借鉴作用。     

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。     

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.     

附子炮制品及其煎煮液6种生物碱的含量测定

目的从附子炮制品来源、煎煮、配伍等方面评价其的安全性和有效性,为临床用药的提供一定参考依据。方法采用高效液相色谱法同时测定不同来源附子炮制品、单方煎煮和复方煎煮液中6种生物碱含量,综合评价附子炮制品的质量及煎煮、配伍对其有效成分和毒性成分的影响。结果不同来源黑顺片中6种生物碱含量差异较大,单酯型生物碱在0.303 5~1.394 8 mg/g之间,双酯型生物碱的含量在0~0.094 0 mg/g之间;炮附片生物碱含量偏低,单酯型生物碱在0.099 0~0.213 5 mg/g之间;两种炮制品经煎煮后,双酯型生物碱基本消失,单酯型生物碱含量也有所降低,其中黑顺片单煎液的溶出率为58.92%,炮附片单煎液的溶出率为87.87%,其复方煎煮液中单酯型生物碱含量均比单煎液偏高。结论 12批黑顺片中6种生物碱含量均符合药典要求,炮附片中生物碱含量偏低,不合格现象明显,煎煮液中双酯型生物碱含量基本消失,毒性更小,与甘草和干姜配伍后,有增效作用。     

附子的水解程度测定

附子是常用中药，但具有大毒，其剧毒成分为双酯型乌头碱，临床上常用水解法（即长时间煎）减缓毒性，但水解程度难于控制。本文采用异羟肟酸铁法测定其酯型乌头碱含量，可衡量乌头碱水解程度，取得满意结果。     

基于UPLC-TOF/MS分析人参附子配伍减毒的物质基础

利用超高效液相色谱-飞行时间质谱联用技术(UPLC-TOF/MS)分析人参附子药对配伍减毒的物质基础,从化学成分层次阐释其配伍减毒机制。基于UPLC-TOF/MS建立人参附子药对配伍后生物碱类成分的化学指纹图谱,通过主成分分析法和正交偏最小二乘判别法分析药对配伍在合煎过程中的生物碱类成分的含量变化,找出差异变化显著的化学成分。结果表明,正离子模式时人参附子药对合煎液中次乌头碱、去氧乌头碱的含量明显降低,而苯甲酰中乌头原碱、苯甲酰次乌头原碱和去乙酸中乌头原碱等含量升高。人参附子药对配伍应用时双酯型二萜生物碱的含量明显降低,而单酯型二萜生物碱的含量明显升高,这可能是人参附子药对配伍减毒作用的物质基础。     

不同煎煮时间对熟附子生物碱的影响

目的 通过测定熟附子不同煎煮时间的生物碱含量,确定最佳的煎煮时间,为规范化的煎煮工艺提供参考.方法 采用HPLC和UV方法分别测定熟附子煎煮液中6种生物碱和总生物碱的含量,并以此含量为指标综合评价其煎煮工艺.结果 熟附子主要含有单酯型生物碱和微量的双酯型生物碱,煎煮0.5h后,3种双酯型生物碱完全消失,而煎煮1h...     

不同煎煮时间对熟附子生物碱的影响

目的通过测定熟附子不同煎煮时间的生物碱含量,确定最佳的煎煮时间,为规范化的煎煮工艺提供参考。方法采用HPLC和UV方法分别测定熟附子煎煮液中6种生物碱和总生物碱的含量,并以此含量为指标综合评价其煎煮工艺。结果熟附子主要含有单酯型生物碱和微量的双酯型生物碱,煎煮0.5 h后,3种双酯型生物碱完全消失,而煎煮1 h后,总单酯型生物碱和总生物碱均达到了峰值,分别为:2.900 1 mg/g和4.832 0 mg/g,随着煎煮时间的延长,单酯型生物碱和总生物碱的含量均逐渐下降。结论熟附子的最佳煎煮时间为1 h。     

附子与甘草配伍前后含量的变化

目的研究附子与甘草配伍前后产生的化学成分的动态变化现象,探讨附子甘草配伍减毒增效的物质基础。方法用高效毛细管电泳法（HPCE）测定附子、甘草单煎及附子与甘草配伍合煎液中甘草苷、甘草酸、乌头碱的含量的动态变化情况。结果附子甘草配伍后乌头碱、甘草苷、甘草酸含量均比单煎时低。结论甘草苷、甘草酸、乌头碱等化学成分的变化是附子甘草配伍能减毒增效的重要物质基础。本实验从组分合和的角度,肯定了附子甘草组分配伍的合理性与科学性。     

The effect of planting depth,disbudding, and root pruning on root weight and aconite alkaloid content of the tuberous root of aconite

The tuberous root of aconite, Aconiti tuber, contains tonic but poisonous aconite alkaloids, such as aconitine, hipaconitine and mesaconitine. The standardization of the cultivation protocols must be established, since the quality of the crude drug, especially the content of aconite alkaloids, is influenced by cultivation conditions. Here, we report the effects of planting depth, disbudding and root pruning on the content of the aconite alkaloids as well as on the weight of the root tuber of aconite. The daughter root tubers of shallow-planted plants showed a significantly higher content of aconitine, hipaconitine and sum of aconite alkaloids per unit weight than those of deeply planted root tubers (control group), while the average weight per root tuber was lower than that of the control group. The total weight of the daughter root tubers per plant of the disbudding group was significantly higher than that of other groups, including the root-pruning group whose average weight per tuber was the highest. In conclusion, disbudding is effective in achieving both higher content of aconite alkaloids and heavier total weight of the root tuber.     

乌头中主要生物碱的高效液相色谱测定

用高效液相色谱分离、测定乌头及附子中一些主要生物碱。生药的乙醚提取物在十八烷基键合相柱上,用甲醇—水—氯仿—三乙胺(70:30:2:0.1)作流动相,中乌头碱、乌头碱及次乌头碱与其它杂质能很好分离。以β-甲基萘作内标化合物,用峰高比测定各生物碱含量。曾测定了不同品种、不同产地及不同加工炮制方法的一些样品,其生物碱组成及比例相差较大。     

Mesaconitine-induced relaxation in rat aorta: involvement of Ca2+ influx and nitric-oxide synthase in the endothelium

Aconiti tuber, roots of aconite (Aconitum japonicum), is an oriental herbal medicine used for centuries in Japan and China to improve the health of persons with a weak constitution and poor metabolism. We investigated the effects of mesaconitine, one of the aconite alkaloids in Aconiti tuber, on the contraction and free intracellular Ca2+ concentration ([Ca2+]i) level in isolated rat thoracic aorta. Mesaconitine at 30 microM inhibited 3 microM phenylephrine-induced contraction in the endothelium-intact, but not endothelium-denuded, aortic rings. The effect of mesaconitine was dependent on external Ca2+ concentrations. The relaxation induced by mesaconitine was abolished by N(omega)-nitro-L-arginine methyl ester (0.1 mM, an inhibitor of nitric-oxide synthase), as well as the relaxation induced by acetylcholine. Acetylcholine induced relaxation in two phases in our conditions; the initial phase was transient and external Ca2+ -independent, and the second phase was sustained and external Ca2+ -dependent. Treatment with 100 nM thapsigargin, which depleted intracellular Ca2+ stores, inhibited acetylcholine-induced, but not mesaconitine-induced, relaxation. Mesaconitine increased the [Ca2+]i level in endothelial cells by influx of Ca2+ from extracellular spaces. These findings suggest that mesaconitine-induced Ca2+ influx and activation of nitric-oxide synthase in endothelial cells and, thus, induced vasorelaxation in rat aorta.     

Intestinal transport of pure diester-type alkaloids from an aconite extract across the Caco-2 cell monolayer model

Aconitine (AC), mesaconitine (MA), and hypaconitine (HA) are the active alkaloids identified in aconite tuber, an important traditional Chinese medicine. The study is aimed to investigate their intestinal transport profiles and potential interaction during the intestinal absorption using the Caco-2 cell monolayer model. All three alkaloids had good permeability with P(app) values greater than 1 × 10 (-6) cm · s (-1). However, AC, MA, and HA in a mixture and as an extract, in both cases with the same content of alkaloids, showed higher transport efficiency in the apical to basolateral, and lower transport efficiency in the basolateral to apical directions. Digoxin, as a P-glycoprotein (P-gp) substrate, was substantially effluxed in the basolateral to apical direction but inhibited by the three alkaloids. Furthermore, the backwards transport of MA and HA was inhibited by the P-gp inhibitor verapamil. These observations indicated that the three alkaloids may not only be P-gp inhibitors but also its substrates; they interact with each other and can potentially enhance their own bioavailability when taken concomitantly.     

Causes and prevention of herb-induced aconite poisonings in Asia

The recent reports from Hong Kong, Taiwan, China and Korea were reviewed to determine the causes and prospects for prevention of herb-induced aconite poisonings. The contributory factors included overdose (use of greater than the recommended doses), faulty processing (after harvest and during decoction), use of tincture (herbal medicinal wine), use of crude aconite roots (for preparing decoction, proprietary medicines and tincture), lack of standardisation in processing of aconite roots and preparation of tincture and proprietary medicines, unsupervised use of aconite roots and contamination or mix-up with aconite roots. As tincture (herbal medicinal wine) made from aconite roots contains a much larger amount of Aconitum alkaloids, the public should be strongly discouraged from making their own and taking it by mouth. Aconite roots should only be used after post-harvest processing and proper decoction. The public should be educated on the hazards from unsupervised use and improper decoction of processed aconite roots. There should be regular publicity measures to promote awareness among the herbalists and to publicise the risk of serious cardiotoxicity if the recommended doses of processed aconite roots are exceeded. The processing of aconite roots and their proprietary preparations should be standardised. Quality control of processed aconite roots and their proprietary preparations should be strengthened. National reporting or monitoring systems can be used to identify the causes of aconite poisonings and assess the impact of preventive measures.     

中药附子质量控制研究

目的HPLC法同时测定附子饮片（生品和炮制品）中3种双酯型生物碱含量,达到控制附子质量及指导临床安全用药的目的。方法以附子饮片（生品和炮制品）为研究对象,采用Agilent Extend—C18（4．6mm×150ram,5lxm）色谱柱,以乙腈-40mmol·L^-1乙酸铵（氨水调pH10．0）为流动相,建立乌头碱、新乌头碱和次乌头碱的含量测定方法,采用外标法测定药材中3种生物碱的含量,并比较各样品之间的差异。结果3种生物碱分离度良好,回收率均在95％－105％。黑顺片中乌头碱、新乌头碱及次乌头碱的含量与其余五批生附片及另一批白附片相比,差别较大。白附片与生附片之间差异较小。同一产地的5批生附片中各生物碱的含量亦存在差异。结论通过同时控制乌头碱、新乌头碱和次乌头碱的含量来控制附子饮片（生品和炮制品）的质量较为合理有效,且简单易行。     

Vasorelaxing effect of mesaconitine,an alkaloid from Aconitum japonicum,on rat small gastric artery: possible involvement of endothelium-derived hyperpolarizing factor

Aconiti tuber, roots of aconite (Aconitum japonicum), has been used for centuries in Japan and China to increase peripheral body temperature. We previously reported that mesaconitine, an alkaloid from Aconitum japonicum, elicits endothelium-dependent and nitric oxide-mediated relaxation in isolated rat aorta. In the present study, we investigated the effect of mesaconitine on isolated rat small gastric arteries. Mesaconitine elicited a concentration-dependent (10, 30, 100 microM) vasorelaxation in isolated rat gastric artery precontracted with norepinephrine, which was resistant to N(omega)-nitro-L-arginine (L-NNA) (an inhibitor of nitric oxide synthase) and indomethacin (an inhibitor of cyclooxygenase). The L-NNA- and indomethacin-resistant relaxation by mesaconitine was mainly endothelium-dependent, inhibited by high K+ (30 mM), and inhibited by a combination of Ca2+-dependent K channel blockers, charybdotoxin and apamin. The relaxation by mesaconitine was proportional to the external Ca2+ concentration. These results suggest that mesaconitine elicits vasorelaxation of isolated rat small gastric artery mainly via release of endothelium-derived hyperpolarizing factor.     

中国乌头的研究Ⅶ.乌头生物碱的薄层层析

本文选择七种已知结构的生物碱,用不加粘合剂氧化铝薄层层析法的桑件进行了研究,结果表明此法完全可应用于乌头生物碱的检定.其较好条件是细度小于150号筛孔,活性Ⅳ级碱性氧化铝为吸附剂;石油醚-乙醚(1:10)为展开剂和以碘蒸气为显色剂.将此薄层层析条件应用到从中国乌头中分出的新生物碱检查时,亦获得良好的结果.