安睡伴侣软胶囊中丁香酚含量测定方法研究

安睡伴侣软胶囊由纯天然传统藏药(包括丁香、肉豆蔻、胡椒、阿魏等6味药材)组方，经超临界CO2萃取其中的挥发油等活性成分后精制而成，具有安神、催眠、抗惊厥等保健功效，无毒副作用。丁香酚是安睡伴侣软胶囊中稳定的、有代表性的功效成分，目前测定药材药品中丁香酚含量的方法主要有液相色谱法(HPLC)和气相色谱法(GC)等。本试验建立了安睡伴侣软胶囊中丁香酚含量的气相色谱和液相色谱测定方法，采用这2种方法测试了5个批次的安睡伴侣软胶囊样品，并将2种方法的测定结果进行了对比。经数据统计分析(配对t检验)显示，HPLC法和GC法所得结果没有显著性差异，2种方法均可满足常规检验要求，都可以作为本制剂的质量检验方法。

四川西北地区川西獐牙菜指纹图谱研究（Ⅰ）

目的：建立川西獐牙菜药材甲醇溶解部分的HPLC指纹图谱。方法：反相高效液相色谱法，流动相甲醇．水(水中含0．02％的磷酸)，梯度洗脱，流速1．0mL·min^-1，检测波长260m，柱温30℃。结果：精密度、重复性、稳定性试验中共有峰峰面积、保留时间的RSD均小于5．0％；不同产地川西獐牙菜的相似度大于0．805。结论：该方法简便、实用、可靠，可作为川西獐牙菜药材的质量标准。

高效液相色谱／质谱分析抱茎獐牙菜提取物中的苷性成分

采用高效液相色谱／质谱法（HPLC／MS）分析抱茎獐牙菜提取物中5种苷性成分。在C18柱上，以甲醇（A：含20％水）和水（B：含10％甲醇）为流动相，流速1mL／min，线性梯度洗脱B从100％到0％，35min，液相色谱-质谱质联用（LC／MS），大气压化学电离源（APCI），对其中5种苷性成分进行定性鉴定。经HPLC／APCIMS分析确证，抱茎獐牙菜提取物中含有獐牙菜苦苷（swertiamarin）、龙胆苦苷（gentiopicroside）、獐牙菜苷（sweroside）、异红草苷（isoorientin）和獐牙菜山酮苷（swertianolin）。采用外标法定量，回收率分别为98．3％、106．7％、92．3％、88．2％和107．3％，该方法简便、快速、准确。

9种“藏茵陈”原植物中的7种有效化学成分研究

目的 研究藏药市场上9种“藏茵陈”植物中有效化学成分，根据所含成分的差异及其生理活性，确定这些植物的药用价值。方法 采用HPLC法测定7种成分的量。结果 9种植物的有效成分差异很大。结论 针对不同的化学成分用途，选择不同的植物，有效利用资源。[著者文摘]

Comparative Study of Contents of Four Tropane Alkaloids in Cultural and Wild Anisodus tanguticus

To assess the medicinal value of cultural Anisodus tanguticus,the contents of four bioactive tropane alkaloids,anisodine,anisodamine,scopolamine and atropine,in cultural and wild materials were determined by the HPLC method.The results showed that content of each alkaloid in the aboveground parts of cultural and wild samples was lower than that in roots,and this explained why it was not the whole plant but the root that was used as medicinal materials.The content of each alkaloid in the roots of one-year cultural material was lower than that in the two-year plants.The discrepancy of the total of four alkaloids between one-year and wild plants is not significant.Moreover,the total of four alkaloids,and the contents of anisodine,scopolamine,and atropine in two-year plants were higher than those in wild plant.Thus there is medicinal value in the cultivated A.tanguticus as well as wild A.tanguticus,especially in the two-year cultural A.tanguticus.

栽培花锚中抗肝炎活性成分的含量在不同生长期变化的研究

研究了花锚中去甲氧基花锚甙和花锚甙的含量随着不同生长期的变化趋势,为药材的合理栽培和采收提供科学依据.RP-HPLC法,使用VP-ODS C18柱,流动相为乙腈∶磷酸∶水(1‰),梯度洗脱程序:0～5.00 min乙腈的体积分数(以下同)为15%、5.01～14.00 min由15%增至25%、14.01～30.00 min由25%增至40%,流速为1 mL/min,柱温25℃,检测波长:254 nm.花锚甙和去甲氧基花锚甙、在花锚全草中的含量在不同生长期有明显变化.

藏药椭圆叶花锚中抗肝炎有效成分的含量测定

运用RP-HPLC建立了花锚中青兰苷、去甲氧基花锚苷和花锚苷的含量分析方法,为栽培花锚替代野生花锚入药提供一定的科学依据.花锚苷、去甲氧基花锚苷和青兰苷分别在0.68～3.40 μg(r=0.999 8)、0.36～1.80 μg(r=0.999 0)和0.80～4.00 μg(r=0.997 9)有良好线性关系,平均回收率分别为100.39% (RSD=2.60%)、99.79% (RSD=3.55%)和100.71% (RSD=2.19%).栽培花锚中花锚苷和去甲氧基花锚苷的含量(抗肝炎活性成分)和在野生花锚中的含量相比无明显差别,可以初步证明栽培花锚可以替代野生花锚入药.

陇西地区蒙古黄芪不同密度栽培试验研究

在陇西首阳黄芪人工栽培基地按照海拔、地形和地貌条件的不同,各选取666.67 m2试验样地,按照不同密度设计株行距,统一种苗和田间管理技术进行栽培试验,收获期统一测定黄芪产量和个体特征,利用HPLC方法对不同密度栽培黄芪甲甙含量进行测试.研究结果表明,从黄芪的外观等级、经济效益和黄芪甲甙含量等指标评价,最佳栽培密度为20 cm×20 cm.

山莨菪植物体内4种莨菪烷类生物碱含量的变化

为了对山莨菪植物资源进行可持续利用，采用高效液相色谱（HPLC）技术对山莨菪植物体内4种莨菪烷类生物碱同时进行了定量测定，将4种生物碱很好地分离开来，大大缩短了出峰时间；并对这4种生物碱含量随着物候的变化进行了分析研究。研究结果表明：4种生物碱含量与物候的关系呈抛物线变化，樟柳碱在山莨菪植物体内含量最高，地上部分4种生物碱均在花盛期含量最高，地下部分不同的生物碱含量最高点则对应不同的物候期。

高寒藏药--抱茎獐牙菜组织培养研究及其愈伤组织有效成分的测定

从抱茎獐牙菜的胚轴、幼叶及未成熟种子诱导出愈伤组织并再生植株,试验选用MS、B5 和N6 三种培养基,其中以附加2. 4 - D3. 0mg/ L + 6 - BA0. 5mg/ L的MS 培养基诱导率最高;以附加6 - BA0. 5mg/ L + NAA 0. 2mg/ L 的MS 培养基分化苗频率最高;以附加2. 4 - D2. 0mg/ L + 6 - BA0. 5mg/ L 的MS 培养基愈伤组织的生长最好。结果表明,外植体,培养基,激素等对愈伤组织诱导、继代和分化均有明显影响。采用高压液相色谱法(HPLC) 测定抱茎獐牙菜愈伤组织中齐墩果酸含量的结果表明,愈伤组织中齐果墩酸含量因培养基、继代培养时间的不同而有所差异。

Simultaneous determination of monoamine and amino acid neurotransmitters in rat endbrain tissues by pre-column derivatization with high-performance liquid chromatographic fluorescence detection and mass spectrometric identification

A sensitive and efficient method for simultaneous determination of glutamic acid (Glu), gamma-amino-butyric acid (GABA), dopamine (DA), 5-hydroxytryptamine (5-HT) and 5-hydroxyindole acetic acid (5-HIAA) in rat endbrains was developed by high-performance liquid chromatography (HPLC) with fluorescence detection and on-line mass spectrometric identification following derivatization with 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC). Different parameters which influenced derivatization and separation were optimized. The complete separation of five neurotransmitter (NT) derivatives was performed on a reversed-phase Hypersil BDS-C-18 column with a gradient elution. The rapid structure identification of five neurotransmitter derivatives was carried out by on-line mass spectrometry with electrospray ionization (ESI) source in positive ion mode, and the BCEOC-labeled derivatives were characterized by easy-to-interpret mass spectra. Stability of derivatives, repeatability, precision and accuracy were evaluated and the results were excellent for efficient HPLC analysis. The quantitative linear range of five neurotransmitters were 2.441-2 x 10(4) nM, and limits of detection were in the range of 0.398-1.258 nM (S/N = 3:1). The changes of their concentrations in endbrains of three rat groups were also studied using this HPLC fluorescence detection method. The results indicated that exhausting exercise could obviously influence the concentrations of neurotransmitters in rat endbrains. The established method exhibited excellent validity, high sensitivity and convenience, and provided a new technique for simultaneous analysis of monoamine and amino acid neurotransmitters in rat brain. (C) 2008 Elsevier B.V. All rights reserved.

Mass spectrometric identification of multihydroxy phenolic compounds in Tibetan herbal medicines

A highly selective and accurate method based on derivatization with dansyl chloride coupled with liquid chromatography-mass spectrometry has been developed for identification of natural pharmacologically active phenolic compounds in extracts of Lomatogonium rotatum plants (Tibetan herbal medicine) obtained by solid-phase extraction. The number of hydroxyl groups on the dansylated phenols was estimated by LC-MS-MS analysis in positive-ion mode. Dansyl derivatization of the compounds introduced basic secondary nitrogen into the phenolic core structures and this was readily ionized when acidic HPLC mobile phases were used. MS fragmentation of the derivatives generated intense protonated molecular ions of m/z [MH](+) (phenol aglycones were transformed into the corresponding free phenols by cleavage of an aglycone bond). Collision-induced dissociation of the protonated molecule generated characteristic product ions of m/z 234 and 171 corresponding to the protonated 5-(dimethylamino)naphthalene sulfoxide and 5 -(dimethylamino) naphthalene moieties, respectively. Selected reaction monitoring based on the m/z [MH](+) to 234 and 171 transitions was highly specific for these phenolic compounds. Characteristic ions with m/z values of [MH - 234](+), [MH 2 x 234](+), and [MH - 3 x 234](+) were of great importance for estimation of the presence of multihydroxyl groups on the phenolic backbone.

Fingerprinting quality control of Qianghuo by high-performance liquid chromatography-photodiode array detection

A novel, simple and accurate fingerprint method was developed using high-performance liquid chromatography-photodiode array detection (HPLC-DAD) for the quality control of Qianghuo, a Tibetan folk and Chinese herbal medicine used as a diaphoretic, an antifebrile and an anodyne. For the first time, the feasibility and advantages of employing chromatographic fingerprint were investigated and demonstrated for the evaluation of Qianghuo by systematically comparing chromatograms of aqueous extracts with the professional analytical software recommended by State Food and Drug Administration (SFDA). Our results revealed that the chromatographic fingerprint combing similarity evaluation could efficiently identify and distinguish raw herbs of Qianghno from different sources and different species. The effects on Notopterygium forbesii Boiss (Apiaceae) chromatographic fingerprints resulted from collecting locations, harvesting time were also examined. (c) 2006 Elsevier lrelanc Ltd. All rights reserved.

Enhancement of atmospheric pressure chemical ionization for the determination of free and glycine-conjugated bile acids in human serum

A highly sensitive and accurate method based on the precolumn derivatization of bile acids (BA) with a high ionization efficiency labeling reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl-benzenesulfonate (BDEBS) coupled with LC/MS has been developed. After derivatization, BA molecules introduced a weak basic nitrogen atom into the molecular core structure that was readily ionized in commonly used acidic HPLC mobile phases. Derivatives were sufficiently stable to be efficiently analyzed by atmospheric pressure chemical ionization (APCI)-MS/MS in positive-ion mode. The MS/MS spectra of BA derivatives showed an intense protonated molecular ion at m/z [M + H](+). The collision-induced dissociation of the molecular ion produced fragment ions at [MH - H2O](+), [MH - 2H(2)O](+), [MH - 3H(2)O](+). The characteristic fragment ions were at m/z 320.8, 262.8, and 243.7 corresponding to a cleavage of N - CO, O - CO, and C - OCC, respectively, and bonds of derivatized molecules. The selected reaction monitoring, based on the m/z [M + H]+ -> [MH - H2O](+), [MH - H2O](+), [MH - 2H(2)O](+), [MH-3H(2)O](+), 320.8, 262.8, and 243.7 transitions, was highly specific for the BA derivatives. The LODs for APCI in a positive-ion mode, at an S/N of 5, were 44.36-153.6 fmol. The validation results showed high accuracy in the range of 93-107% and the mean interday precision for all standards was < 15% at broad linear dynamic ranges (0.0244-25nmol/mL). Good linear responses were observed with coefficients of > 0.9935 in APCI/MS detection. Therefore, the facile BDEBS derivatization coupled with mass spectrometric analysis allowed the development of a highly sensitive and specific method for the quantitation of trace levels of the free and glycine-conjugated BA from human serum samples.

2-(2-Phenyl-1H-phenanthro-[9,10-d]imidazole-1-yl)-acetic acid (PPIA) and its application for determination of amines by high performance liquid chromatography with fluorescence detection and identification with mass spectroscopy/atmospheric pressure chemical ionization

A simple, sensitive, and mild method for the determination of amino compounds based on a condensation reaction with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC-HCI) as the dehydrant with fluorescence detection has been developed. Amines were derivatized to their acidamides with labeling reagent 2-(2-phenyl-1H-phenanthro-[9,10-d]imidazole-1-yl)-acetic acid (PPIA). Studies on derivatization conditions indicated that the coupling reaction proceeded rapidly and smoothly in the presence of a base catalyst in acetonitrile to give the corresponding sensitively fluorescent derivatives with an excitation maximum at lambda(ex) 260nm and an emission maximum at lambda(em) 380nm. The labeled derivatives exhibited high stability and were enough to be efficiently analyzed by high-performance liquid chromatography. Identification of derivatives was carried out by online post-column mass spectrometry (LC/APCI-MS/MS) and showed an intense protonated molecular ion corresponding m/z [MH](+) under APCI in positive-ion mode. At the same time, the fluorescence properties of derivatives in various solvents or at different temperature were investigated. The method, in conjunction with a gradient elution, offered a baseline resolution of the common amine derivatives on a reversed-phase Eclipse XDB-C-8 column. LC separation for the derivatized amines showed good reproducibility with acetonitrile-water as mobile phase. Detection limits calculated from 0.78 pmol injection, at a signal-to-noise ratio of 3, were 3.1-18.2 fmol. The mean intra- and inter-assay precision for all amine levels were < 3.85% and 2.11%, respectively. Excellent linear responses were observed with coefficients of > 0.9996. The established method for the determination of aliphatic amines from real wastewater and biological samples was satisfactory. (c) 2006 Elsevier B.V. All rights reserved.