苦地丁中六种异喹啉生物碱的薄层分离和光密度法测定

在碱性硅胶G薄层上用己烷—氯仿—甲醇(8∶2∶0.25及4∶5∶1)两种溶剂系统,分别展开分离了苦地丁中六种异喹啉生物碱——普洛托品(Ⅰ)、右旋异紫堇灵(Ⅱ)、右旋紫堇灵(Ⅲ)、四氢黄连碱(Ⅳ)、乙酰紫堇灵(Ⅴ)及苦地丁5号(Ⅵ)。用改良Dragendorff试剂显色后,薄层光密度法测定。各生物碱的线性范围为0.5～3.0μg,平均变异系数为1.76%,显色后24小时内稳定。用本法可测定苦地丁及紫堇属多种植物,苦地丁总碱及片剂中异喹啉生物碱的含量。     

RP-HPLC法测定苦地丁及其复方制剂中紫堇灵的含量

采用反相高效液相色谱法测定苦地丁及其中药制剂感冒清热颗粒中紫堇灵的含量。色谱柱为APEX -ODS柱 ;流动相为甲醇 1 5mmol/L磷酸盐缓冲液 pH 6 70 ( 70∶30 ) ;流速为0 8mL/min ;检测波长为 2 89nm ;采用外标两点法进行定量。在紫堇灵为 6 9～ 1 1 0 4 μg/mL范围内线性关系良好 ,相关系数r=0 9999。方法精密度RSD分别为 1 5 %和 0 8% ,回收率为 97 3%和 97 5 %。方法适用于不同产地苦地丁及 3种不同剂型感冒清热颗粒中的紫堇灵含量。     

紫堇灵、乙酰紫堇灵及原鸦片碱对小鼠实验性肝损伤的保护作用

研究了紫堇灵、乙酰紫堇灵及原鸦片碱对小鼠实验性肝损伤的保护作用及其作用机理。小鼠预先分别ig紫堇灵、乙酰紫堇灵或原鸦片碱50及100mg·kg^-12次,对CCl₄、硫代乙酰胺、扑热息痛所致的小鼠肝损伤均有保护作用,使SGPT显著降低,肝病理损伤程度减轻。此3种成分在体外均能抑制CCl₄引起的肝微粒体脂质过氧化及CCl₄转化为CO。在上述实验中乙酰紫堇灵的作用均强于另外两种成分。另外,此3种成分对肝药酶有先抑制后诱导作用。     

紫堇属植物生物碱的化学——青海高原四种紫堇属植物生物碱的化学研究

中草药夏天无(Corydalis decumbens)和苦地丁(Corydalis bungeana)的总生物碱,经过临床试用,已作为治疗心血管系统疾病的药物。这两药都是紫堇属植物,且主要含异喹啉类生物碱,根据这些资料,拟从这属植物中寻找治疗心血管系统疾病的药物。我们首先选择了直茎紫堇(Corydalis stricta Steph.)、暗绿紫堇(C.melanochlora Maxim.)、条裂紫堇(C.lineariodes Maxim.)、短尖紫堇(C.mucronifera Maxim.)四种植物,利用离子交换树脂     

中药苦地丁、牵牛子的荧光学测定

目的：探讨中药苦地丁、牵牛子荧光学测定方法。方法：将苦地丁、牵牛子溶液分别加入硅胶G板，经荧光分光光度计扫描薄层板，根据面积及质量的关系制作标准曲线，代入线性方程计算含量。结果：苦地丁中紫堇灵含量、牵牛子中咖啡酸含量经荧光检测三次所得结果差异无统计学意义（P〉0.05），提示荧光测定值稳定性及准确性较高。结论：应用荧光学测定苦地丁、牵牛子中有效成分准确性较高，是临床确定药物有效成分及对药材进行准确鉴别的有效方法。     

异紫堇啡碱对家兔窦房结及豚鼠心室肌动作电位的作用

用细胞内固定微电极技术观察了异紫堇啡碱(isocorydine)对家兔窦房结电活动、豚鼠心室肌动作电位及Ba²⁺诱发的豚鼠心室肌自发电活动的作用。结果表明,异紫堇啡碱能使窦房结细胞APA,SP₀,SP₄减小、APD₉₀延长、自律性降低。3 μM异紫堇啡碱能使心室肌细胞的APD₅₀,APD₉₀和ERP延长,300μM异紫堇啡碱则使心室肌细胞APD₅₀和APD₉₀缩短,但不缩短ERP,使ERP相对延长。300μM异紫茧啡碱亦能明显抑制Ba²⁺诱发的心室肌自发电活动。     

矮紫堇中紫堇灵和乙酰紫堇碱含量测定及体外抗氧化活性评价

目的：建立同时测定矮紫堇中紫堇灵和乙酰紫堇碱含量的方法,并测定矮紫堇粗提物的体外抗氧化活性。方法：采用高效液相法测定矮紫堇中紫堇灵和乙酰紫堇碱含量。采用1,1-二苯基-2-苦肼基（DPPH·）清除体系、羟基自由基（·OH）清除体系和NaNO₂清除体系评价矮紫堇粗提物的体外抗氧化活性。结果：矮紫堇中紫堇灵和乙酰紫堇碱的含量分别为2.49,4.36 mg·kg^-1。矮紫堇粗提物对羟基自由基·OH的清除率随浓度升高而增强,清除率最高可达85.21%,而对DPPH·和NaNO₂清除率最高,分别为90.86%,90.66%。结论：本实验建立了同时测定矮紫堇中紫堇灵和乙酰紫堇碱含量的高效液相色法,本方法操作简单,重复性好。矮紫堇粗提物具有显著的抗氧化活性,并呈现出量效关系。     

紫堇灵、乙酰紫堇灵及原鸦片碱对小鼠实验性肝损伤的保护作用

研究了紫堇灵、乙酰紫堇灵及原鸦片碱对小鼠实验性肝损伤的保护作用及其作用机理。小鼠预先分别ｉｇ紫堇灵、乙酰紫堇灵或原鸦片碱５０及１００ｍｇ·ｋｇ－１２次，对ＣＣｌ４、硫代乙酰胺、扑热息痛所致的小鼠肝损伤均有保护作用，使ＳＧＰＴ显著降低，肝病理损伤程度减轻。此３种成分在体外均能抑制ＣＣｌ４引起的肝微粒体脂质过氧化及ＣＣｌ４转化为ＣＯ。在上述实验中乙酰紫堇灵的作用均强于另外两种成分。另外，此３种成分对肝药酶有先抑制后诱导作用。     

苦地丁及其伪品的鉴别

苦地丁为常用中药,具清热解毒,凉血清肿的功能.苦地丁之名收载于《中华人民共和国药典1977年版》,以区别于他种地丁,并规定原植物为罂粟科植物紫堇Corydalis bu-ngeana Turcz.的干燥全草.历来无伪品.1991年5月笔者在安国药材市场上发现一种假     

康脑灵对大鼠缺氧状态下脑保护作用的定量脑电图

目的：通过皮层脑电(ECoG)的定量分析评价康脑灵(Kang Nao Lin, KNL)对缺氧状态下大鼠的脑保护作用。方法：用定量方法分析在体脑电在正常供气与停止呼吸情况下的变化，判断药物的脑保护作用。结果：ig康脑灵可以显著延长停止供气后脑电平均振幅到达最低点的时间，促进重新供气后脑电活动的完全恢复。 这种作用在0.63g.kg^-1和1.26g.kg^-1间呈剂量依赖性，与对照组有极显著差异，与氟桂嗪组(2.5mg.kg^-1)无显著性差异。其中康脑灵1.26g.kg^-1组在缺氧2 min时脑电不完全消失。ig康脑灵可以不同程度地降低皮层脑电的平均振幅。结论：ig KNL在一定剂量范围内有明显的抗脑缺氧的作用。     

安徽贝母生物碱的分离和结构鉴定

从安徽贝母(fritillaria anhuiensis S.C.Chen et S.F.Yin)鳞茎中分离出四个生物碱Ⅰ,Ⅱ,Ⅲ及Ⅳ,通过光谱数据及与标准品对照,碱Ⅰ为浙贝乙素(verticinone,peiminine),碱Ⅱ为贝母辛(peimisine),碱Ⅲ为异浙贝甲素(isoverticine)。碱Ⅳ是一种新的生物碱,定名为皖贝甲素(wanpeinine A)。根据IR,MS,¹HNMR以及¹³CNMR光谱解析和衍生物制备,推定其结构为5α,14α,22β-cevanine-3β,6α,20β-triol。     

麦角单菌核中麦角生物碱的薄层分离及荧光光密度法测定

本文介绍了麦角单菌核中麦角新碱、麦角酰胺、麦角胺及麦角克碱的薄层分离及荧光光密度法定量的分析方法。本法快速、灵敏,检出限度为1×10^-8g,四种生物碱的平均变异系数为6.10%,层离后的薄层斑点避光保存可稳定24小时。本法为探索麦角单个菌核问含麦角生物碱的质和量的变化以及为定向选育菌种提供了可能性。     

三尖杉酯类生物硷对小鼠白血病P388及L615蛋白质生物合成的影响

本文初步观察了四种三尖杉酯硷对³H-DL-亮氨酸、³H-L-门冬酰胺、¹⁴C-DL-苯丙氨酸掺入小鼠白血病P₃₈₈腹水细胞及L₆₁₅。脾组织蛋白质的作用。实验结果表明,在等毒性剂量(1/5 LD₅₀)下,一次腹腔(或静脉)给药,四种酯硷对P₃₈₈腹水细胞及L₆₁₅脾组织蛋白质生物合成抑制明显,作用较快,并具有相似的时间—反应曲线。一般在24小时基本恢复。     

Cytotoxicity,Cell Cycle Kinetics and Morphonuclear-induced Effects of Vinca Alkaloid Anticancer Agents

The effects of four Vinca alkaloids (vinblastine, vincristine, vindesine and vinorelbine) on three neoplastic cell lines (the MXT mouse mammary cell line and the T24 and J82 bladder cell lines) were studied at three biological levels, i.e. cell proliferation, cell cycle kinetics and morphonuclear characteristics. These effects were studied by means of digital cell image analysis on Feulgen-stained nuclei. The aim of the present work was to characterize the effects specifically induced by Vinca alkaloids as compared with those obtained previously with other pharmacological classes of anticancer drugs. The results show that Vinca alkaloids inhibit the cell proliferation of neoplastic cell lines at a concentration of 10^***-8 M except in the case of the J82 cell line, for which only a slowing down of cell proliferation was observed. Concerning the cell cycle kinetics, the results show that the Vinca alkaloids induce an accumulation of cells in the mitosis phase. This accumulation of mitotic cells was maximal after 15 h incubation in the presence of the drugs. A study of the morphonuclear-induced effects of Vinca alkaloids showed that the variance of the optical density (VOD) is strongly influenced by these Vinca alkaloids. The development of the VOD was parallel with the development of the percentage of mitosis; thus, the VOD enabled the Vinca alkaloid-induced effects to be specifically characterized from a morphonuclear point of view. On the other hand, the results show that the mean value of the variance of the optical density was very highly correlated (P < 0***.001) with the efficiency of the Vinca alkaloids in terms of cytotoxicity. In clinical studies, the analysis of the development of this parameter would make it possible to assess the response to chemotherapy in the case of patients treated with Vinca alkaloids.     

十大功劳属部分植物茎中生物碱的高效毛细管电泳法测定

目的：对十大功劳属植物中3种生物碱的含量进行测定,为其内在质量的评价提供科学依据。方法：运用毛细管区带电泳(CZE)模式,以马钱子碱为内标化合物,测定了小檗碱、巴马亭和药根碱的含量。背景缓冲溶液为0.1 mol.L^-1磷酸缓冲液(pH 7.0)与甲醇(2∶1)的混合体系。线性范围：盐酸小檗碱为4.9～498.6 μg.mL^-1(γ=0.9990);盐酸巴马亭为5.0～504.9 μg.mL^-1(γ=0.9996);盐酸药根碱为5.1～505.8 μg.mL^-1(γ=0.9984)。回收率：盐酸小檗碱为96.00%～101.66%;盐酸巴马亭为100.15%～102.97%;盐酸药根碱为96.68%～102.44%。结果：不同品种、不同产地、不同采集时间的植物中的生物碱含量均有较大差别。结论：高效毛细管电泳法是中草药质量控制的一种简便、快速、有效的方法。十大功劳属多种植物生物碱含量较高,可作为新的药用资源。     

紫乌头生物碱的化学研究

从紫乌头根中分出四种新二萜生物碱,分别命名为紫乌生、紫乌生宁、紫乌亭和紫乌定。根据红外光谱、紫外光谱、质谱、氢谱和碳谱以及制备衍生物,证明了前三个生物碱的结构(3,4,5),对紫乌定的结构曾初步推定为(8)⁽¹⁾;后来,经单晶X射线衍射结构分析修正为(9)。此外,作者认为,根据化学结构特点应从C₁₉二萜生物碱中划分出C-4失甲基的一类生物碱,称C₁₈二萜生物碱。     

Microwave assisted solid phase microextraction for extraction and selective enrichment of four alkaloids in lotus leaf

A novel method was developed for extraction and enrichment of the four alkaloids (nuciferine, O-nornuciferine, armepavine and N-nornuciferine) from lotus leaf by coupling microwave-assisted extraction (MAE) with solid phase microextraction (SPME) before ultra-high-performance liquid chromatography (UHPLC) analysis. The influence parameters of MAE and SPME process including solvent-to-materials ratio, extraction solvent, irradiation power, extraction time were investigated. The method exhibited a good linearity within the range of 0.4–100 μg mL^?1 for nuciferine and O-nornuciferine, 0.1–25 μg mL^?1 for armepavine and N-nornuciferine (r² > 0.9990), respectively. Compared with the other extraction method, this method obviously shortens analytical time and enhances extraction efficiency. In addition, the concentration of the four alkaloids (nuciferine, O-nornuciferine, N-nornuciferine and armepavine) included two trace alkaloids were increased by 13, 10, 5 and 12 times after concentration, respectively. It was concluded that the newly MAE-SPME was an efficient method for the extraction and enrichment for alkaloids from herbs.     

Cytotoxicity of alkaloids isolated from Argemone mexicana on SW480 human colon cancer cell line

Context: Argemone mexicana Linn. (Papaveraceae) has been used as traditional medicine in India and Taiwan for the treatment of skin diseases, inflammations, bilious, fever, etc. Some alkaloids of A. mexicana have been screened for their cytotoxicity on different cancer cell lines.

Objective: The study investigates potential cytotoxic effects of alkaloids isolated from aerial part of A. mexicana on SW480 human colon cancer cell line.

Materials and methods: Six alkaloids, 13-oxoprotopine, protomexicine, 8-methoxydihydrosanguinarine, dehydrocorydalmine, jatrorrhizine, and 8-oxyberberine were isolated from the methanol extract of A. mexicana. Cytotoxicity of these alkaloids was studied on SW480 human colon cancer cell line at 1, 25, 50, 75, 100, 125, 150, and 200?µg/mL for 24 and 48?h. Cells were seeded in a 96-well micro-plate at a concentration of 2?×?10⁴ cells per well and MTS assay was performed to assess cytotoxicity in terms of cell viability.

Results: At 200?µg/mL, protomexicine and 13-oxoprotopine showed mild cytotoxicity (～24–28%) whereas dehydrocorydalmine exhibited moderate cytotoxicity (～48%). 8-Oxyberberine was mildly cytotoxic (～27%) at 24?h but was more potent (～76%) at 48?h. Jatrorrhizine and 8-methoxydihydrosanguinarine were most potent (～95–100%) in inhibiting the human colon cancer cell proliferation showing complete reduction in cell viability.

Discussion and conclusion: This is the first study on the effect of these alkaloids on SW480 human colon cancer cell line. This study indicates that some alkaloids of A. mexicana strongly inhibit the cell proliferation in human colon cancer cells, and it might be a basis for future development of a potent chemotherapeutic drug.     

Study on the aconitine‐type alkaloids of Radix Aconiti Lateralis and its processed products using HPLC‐ESI‐MSn

The type and content change of alkaloids of Radix Aconiti Lateralis (Lateral root of Aconitum carmichaeli Debx, an important and popular medicinal herb in Traditional Chinese Medicine) in processing were studied using high performance liquid chromatography‐electrospray ionization‐multi‐stage mass spectrometry (HPLC‐ESI‐MSⁿ). Extract containing alkaloids, which were known to be the main bioactive components of the herb, was prepared by 1% (v/v) hydrochloric acid solution. An HPLC method which can simultaneously separate these alkaloids was established with gradient elution mode. Forty‐eight compounds were structurally identified by employing LC‐MSⁿ techniques; the MSⁿ spectra of most alkaloids displayed a characteristic behaviour of loss of CH₃COOH (60 u), CH₃OH (32 u), C₆H₅COOH (122 u), CO (28 u) and H₂O (18 u). Among them, the fragmentation ion C6H5COOH (122 u) was reported for the first time. By comparison, 22 compounds were found both in the crude materials and the processed products; 17 alkaloids were only found in the processed products and 9 alkaloids,which existed in crude material, could not be detected after processing. In the process of identification, we found new kinds of alkaloids, with protonated molecules at m/z 452, 468, and 482. We called these compounds dehydra‐hypaconine, dehydra‐mesaconine, and dehydra‐aconine, respectively. Copyright © 2012 John Wiley & Sons, Ltd.     

A fragmentation study on four C19-diterpenoid alkaloids by electrospray ionization ion-trap time-of-flight tandem mass spectrometry

High-resolution electrospray ionization ion-trap time-of-flight tandem mass spectrometry (HR-ESI-IT-TOF-MSⁿ) in positive-ion mode was used to determine the accurate masses and fragmentation pathways of four C₁₉-diterpenoid alkaloids, aconitine (1), yunnaconitine (2), crassicauline A (3), and benzoylmesaconine (4). The [M+H]⁺ ions of compounds 1–4 were readily observed in conventional single-stage mass spectrometry. Based on the MS^1–6 analyses, detailed fragmentation rules of the four compounds were proposed. The neutral losses of AcOH, MeOH, H₂O, CO, C₂H₄, PhCOOH and p-OMePhCOOH segments were the characteristic eliminations from the precursor ions due to the presence of acetyl, methoxyl, hydroxyl, N-ethyl, benzoyl and p-methoxyl-benzoyl units in the structures. Benefited from the high resolution of the mass analyzer, the loss of 28 Da corresponding to CO or CH₄ segment in product ions was unambiguously distinguished. The losing sequence of the main substituent groups was summarized as: C(8)-acetyl>C(16)-methotyl>C(15)-hydroxyl>C(6)-methoxyl>C(1)-methoxyl/C(3)-hydroxyl>C(18)-methoxyl>>C(13)-hydroxyl. The sequential loss of (16)-methoxyl moiety and CO (generating from enol–ketone tautomerism) groups could be recognized as the characteristic eliminations for the compounds with C(16)-methoxyl and C(15)-hydroxyl groups simultaneously. The application of HR-ESI-IT-TOF-MSⁿ technique to investigate the fragmentation of C₁₉-diterpenoid alkaloids provided useful information to understand their fragmentation behaviors.