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。     

不同炮制方法对川乌双酯型生物碱含量的影响

目的 研究炮制方法对川乌双酯型生物碱含量的影响。方法将生川乌分别通过药典法、黑豆法和米醋法进行炮制,以RP—HPLC结合梯度洗脱的色谱方法为检测手段,比较分析不同川乌炮制方法对双酯型生物碱（主要为乌头碱、次乌头碱）含量的影响。结果3种炮制方法均可显著降低生川乌中乌头碱、次乌头碱的含量。所建立的色谱方法可定量分析炮制品中双酯型生物碱。结论不同炮制方法对生川乌均能起到减毒作用。     

附片中总灰分、酸不溶性灰分和总无机盐的测定

目的测定附片中总灰分、酸不溶性灰分和总无机盐的含量。方法按2010年版《中国药典》的方法,测定附片中总灰分和酸不溶性灰分;采用EDTA滴定法测定附片中总无机盐的含量。结果附片中总灰分、酸不溶性灰分和总无机盐的含量差异很大,且显著高于生附子,表明附子加工炮制明显增加了总灰分和无机盐的含量,可能是胆巴漂洗不净、褪胆不全所致。结论所用方法简便、快速、准确,可作为附片的质量控制方法。     

高效液相色谱法测定附子及其炮制品中双酯型生物碱的评价

目的 观察高效液相法在附子及其炮制品中双酯型生物碱含量测定的应用及质量控制.方法 进行供试品及对照品溶液制备,色谱柱为Agilent Zorbax Ex tend-C18柱,乙醚超声提取,检测波长为235 nm.考察供试品和（或）对照品溶液的线性关系,HPLC精密度、稳定性、重复性,以及加样回收率试验和样品测定.结果 乌头碱、新乌头碱、次乌头碱分别为Y=4.070×105X＋ 82.50、Y=3.960×105X ＋6.826、Y=3.956×105X＋7.256,线性范围为0.107～ 4.035/μg、0.100～4.078/μg、0.085 ～ 3.505/μg,精密量RSD为0.67％、0.70％、0.39％,稳定性RSD为0.66％、0.45％、0.75％,重复性RSD为1.65％、1.56％和1.76％,加样回收率RSD为97.30％ （1.37％）、97.25％ （1.40％）和97.60％（1.51％）,生附片中双酯型生物碱含量高低顺序为：次乌头碱＞新乌头碱＞乌头碱.炮制品中,盐附子的毒性已经大大降低,而清水黑顺片、熟附片（干片蒸制）、熟附片（鲜片蒸制）中仅有微量的毒性成分,而炮附片中主要毒性成分的含量仍较高.结论 HPLC法在附子与其炮制品中双酯型生物碱测定中具有方法简便快速、灵敏准确,实验结果稳定,重现性好,专属性强,且成本低、效率高的特点,RSD能达到定量的要求,可能是一种较好的附子中双酯型生物碱的提取方法.     

半夏反附子之说不能成立

半夏为天南星科植物半夏[Pinellia ternate(Thunb.)Breit)的块茎。附子为毛茛科乌头属植物乌头(Aconitum Carmichaeli Debx)的子根。临床内服制剂大都是用其炮制品。药典及近代一些中药书中的附子项下均载有:“本品不     

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。     

诃子制草乌的炮制工艺研究

目的 以乌头碱、新乌头碱和次乌头碱的含量为指标,优化得到诃子制草乌的炮制工艺;并尝试用特征图谱的方法,分析炮制品的质量.方法 建立测定草乌中3种乌头碱含量的HPLC法和测定草乌特征图谱的方法,根据《蒙药炮制学》中记载的工艺进行单因素、正交试验和验证试验,优化炮制工艺,同时,对生品和不同炮制品进行特征图谱的分析,考察相似...     

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。     

反相高效液相色谱法测定参附注射液及市售黑附片中乌头碱的含量

目的　测定参附注射液及市售黑附片中乌头碱的含量。方法　用RP HPLC ,流动相为乙腈 醋酸铵缓冲液 (pH10 5 ) ,紫外检测波长 2 3 0nm ,流速 0 8ml·min-1。结果　对照品的线性范围 1 3 9～ 2 2 2 4 μg(r =0 9997) ;市售黑附片中的乌头碱比参附注射液中的高 3倍。结论　方法简便、快速、准确、重现性好 ,可以作为样品的检验方法     

关于《中国药典(95版一部)》附子叙述方法的建议

《中国药典 ( 95版一部 )》 (以下简称“《药典》”)中 ,关于附子的叙述方法 ,我们认为有些不妥 ,主要是其炮制方法没有按《药典》规定栏目顺序介绍 ,显得较零乱。如 :在叙述完附子原植物及药材来源后 ,紧接着分别介绍了“盐附子”、“黑顺片”和“白附片”的炮制方法 ,其中除了“盐附子”可看作制做淡附片的原料外 ,其他两种均已属直接入药的炮制品 (按《药典》格式 ,炮制品应在“炮制”栏目中介绍 )。而在下面的炮制栏目中则只介绍了“淡附片”和“炮附片”的炮制方法。这就使该药炮制方法的介绍过于分散 ,形成了炮制栏目中谈炮制 ,非炮制…     

附子地上部分化学成分研究

目的 研究附子地上部分的化学成分.方法 采用色谱法进行分离,根据理化性质和波谱数据鉴定化合物的结构.结果 分离得到5个化合物,分别鉴定为次乌头碱(1)、10-羟基乌头碱(2)、新乌头碱(3)、乌头原碱(4)和β-谷甾醇(5).结论 之前没有对附子地上部分进行化学成分研究的报道,本研究首次从中分离的到了化合物1～5.     

全固态乌头碱电化学检测器的研制及其在流动注射分析中的应用

报道一种新型结构的全固态乌头碱电化学检测器的研制及其在流动注射分析中的应用。采用流动注射分析法对川乌、草乌及其中成药(小活络丸)中剧毒成分(双酯型生物碱)进行了测试,方法简便快速,结果同光度法接近。本文还提出了电化学法研究乌头碱水解动力学原理。在pH 6.5,温度98℃的条件下测得乌头碱水解成苯甲酰乌头碱的速度常数为1.36×10^-2min^-1。     

双向发酵对川乌指纹图谱及乌头碱类成分含量的影响

目的:观察川乌被19种药用真菌发酵后,乌头碱、新乌头碱、次乌头碱含量的变化及高效液相色谱(HPLC)指纹图谱的差异。方法:将川乌作为"药性基质",在一定的条件控制下,应用双向发酵技术,使其被槐耳、灵芝、红栓等19种药用真菌发酵,产生药材的"药性菌质",并采用反相高效液相色谱法对"药性菌质"和生药材中所含的乌头碱、新乌头碱和次乌头碱进行含量测定,比较其含量的变化及指纹图谱的差异。结果:在一定的时间范围内,大多数的菌株在川乌基质上的适应性良好,菌丝体生长旺盛,如代号A、C、D、E、F、M、Q、V、SW、い等真菌。发酵之后的川乌中乌头碱、新乌头碱及次乌头碱含量与生药材比较多数有明显的降低。结论:双向发酵后多数"药性菌质"中乌头碱、新乌头碱和次乌头碱含量明显降低了,提示双向发酵是生物技术在中医药领域中应用的有益尝试。     

Determination of aconitine-type alkaloids as markers in fuzi (Aconitum carmichaeli) by LC/(+)ESI/MS(3)

LC/(+)ESI/MS(3) was used to determine aconitine, mesaconitine, and hypaconitine as target markers in crude methanol extracts of (i) the raw lateral roots of Aconitum carmichaeli, (ii) roots treated by three different refining processes, and (iii) eight generally available traditional Chinese medicine (TCM) preparations containing fuzi (treated lateral roots of A. carmichaeli). The optimal ionization behavior resulted when using electrospray ionization (ESI) in positive-ion mode with 0.005% TFA as an additive in the mobile phase. The consecutive reaction monitoring (CRM) mode provided additional improvements in selectivity, which was exploited to minimize the noise and interference problems. Employing this approach, aconitine and mesaconitine were found to decompose readily during the refining processes, but hypaconitine remains present at the same content, presumably because of its characteristic chemical structure. Thus, treated and untreated fuzi samples can be distinguished by monitoring the ratio of aconitine and mesaconitine to hypaconitine. The limits of detection (LODs) for these three markers were 0.05, 0.08, and 0.03 ng/ml. The linearity range for the three marker compounds was 0.1-1,000 ng/ml. The analysis time was 12 min per sample.     

An online coupled cell membrane chromatography with LC/MS method for screening compounds from Aconitum carmichaeli Debx. acting on VEGFR-2

An online analytical method coupling high expression vascular endothelial growth factor receptor (VEGFR) cell membrane chromatography (VEGFR-CMC) with high performance liquid chromatography mass spectrometry (LC/MS) for screening and identification of active component from traditional Chinese herb Aconitum carmichaeli Debx. acting on VEGFR-2 was established. Through a 10-port column switcher, factions separated by VEGFR-CMC column (first dimension) were transferred and were adsorbed on an enrichment column. Then, these fractions were sent into LC/MS system (second dimension) immediately and directly for separation and preliminary identification, respectively. Sunitinib malate (SN) was used as positive control, while nifedipine (NF), dexamethasone acetate (DX), methoxyamine hydrochloride (MT) and atenolol (AT) as negative controls. The specification of this VEGFR-CMC-online-LC/MS method was validated by competitive displacement test. As a result, mesaconitine (MSC), aconitine (AC), and hypaconitine (HPC) were identified as the active constituents acting on VEGFR-2. The in vitro inhibition activity of starting extract of Aconitum carmichaeli Debx., MSC, AC, and HPC on HEK293/VEGFR cell viability by MTT test, separately. The in vitro inhibition activity of MSC, AC, and HPC on vascular endothelial growth factor (VEGF) secretion of HEK293/VEGFR cell was tested by VEGF-ELISA assay. The screening results given by the system offered additional exemplification supporting this online coupling method and gave new evidence to the development of anti-tumor drug from natural products.     

川乌氯仿部位的化学成分研究

目的研究川乌氯仿部位的化学成分。方法通过硅胶、碱性氧化铝柱色谱进行分离纯化,并根据理化性质和波谱数据鉴定化合物的结构。结果分离得到9个化合物,分别鉴定为次乌头碱(1)、金色酰胺醇酯(2)、乌头碱(3)、松胞素B2(4)、宋果灵(5)、新乌头碱(6)、多根乌头碱(7)、北乌碱(8)、附子灵(9)。结论化合物2为首次从乌头属植物中分离得到,化合物4为首次从植物界中分离得到。     

Incidence of herb-induced aconitine poisoning in Hong Kong: impact of publicity measures to promote awareness among the herbalists and the public.

BACKGROUND: In Hong Kong 'chuanwu' (the main root of Aconitum carmichaeli) and 'caowu' (the root of Aconitum kusnezoffii) are used by herbalists to treat patients with various musculoskeletal disorders. These aconite roots contain aconitine, mesaconitine and hypaconitine, which are neurotoxins and cardiotoxins. During 1989 to 1991, 31 patients were treated in public hospitals because of poisoning by aconite roots and there were two deaths from ventricular arrhythmias. In late 1991, healthcare officials together with cardiologists held a press conference to warn the public, healthcare professionals and herbalists of the potential toxicity of aconite roots. The risk of ventricular arrhythmias and the need for urgent medical attention were highlighted. An information leaflet was also sent to hospital doctors. Since 1992, the topic was covered periodically in the local newspapers, medical journals and continuing medical education programmes. OBJECTIVE: The objective of this study was to assess the impact of these publicity measures on the incidence of herb-induced aconitine poisoning in the New Territories East, based on the number of admissions to the Prince of Wales Hospital. METHODS: During 1989 to 1993 and 1996 to 1998, all patients admitted to our medical wards because of herb-induced aconitine poisoning were identified by on-going surveillance of medical patients, searching our computerised medical record system and reviewing reports received by the 24-hour Drug and Poisons Information Bureau. RESULTS: The number of hospitalisations due to aconitine poisoning markedly decreased from four to six per year in 1989 to 1991 to one to two per year in 1992 to 1993. The annual incidence of aconitine poisoning showed a marked decrease from 0.49 to 0.69 [overall 0.60, 95% confidence intervals (CI) 0.34 to 0.99] to 0.10 to 0.22 (overall 0.16, 95% CI 0.03 to 0.46) per 100,000 population (p = 0.024). During 1996 to 1998, herb-induced aconitine poisoning remained uncommon, with zero to two hospital admissions per year or an annual incidence of zero to 0.33 (overall 0.17, 95% CI 0.05 to 0.43) per 100,000 population (p = 0.016). DISCUSSION: It is possible that the herbalists could have used smaller doses of 'chuanwu' and 'caowu' than before. Patients could be more compliant with the instructions on how to prepare the herbal decoction at home. However, our experience suggested that publicity measures to promote awareness, among the herbalists and the public, may reduce the incidence of poisoning due to toxic herbs such as aconite roots.     

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.     

四川江油附子生物碱成分的研究

从四川江油附子(Aconitum carmichaeli Debx)中分得六个化合物。经理化常数测定和光谱分析证明,化合物V为新化合物,命名为新江油乌头碱(neojiangyouaconitine)。其余为已知物,分别鉴定为尿嘧啶(Ⅰ),华北乌头碱(Ⅱ),黄草乌头碱(Ⅲ),尼奥灵(Ⅳ),附子亭(Ⅵ)。     

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.