石蒜科生物碱的研究 Ⅲ.紫花石蒜和其他两种石蒜中的生物碱及新生物碱紫花石蒜碱

从紫花石蒜Lycoris squamigera Maxim.中分得19种结晶性生物碱,其中12种生物碱分别证明为lycorine,pseudolycorine,lycorenine,homolycorine,tazettine,norpluviine,galanthamine,epigalanthamine,vittatine,pluviine,lycoramine,hippeastrine;分得一种新生物碱,命名为紫花石蒜碱squamigerine. 紫花石蒜碱,C₁₈H₂₁O₅N,熔点260℃,[α]_D³⁴+165°(氯仿),含有次甲二氧基、氯甲基、甲氧基、羟基和双键各1个。紫花石蒜碱的分子式及官能团均与tazettine相同;熔点,[α]D及R_f值不同。因而推想紫花石蒜碱可能为tazettine的异构体。我们进行了两者的紫外及红外吸收光就业谱的比较,发现它们彼此非常相似。因为紫花石蒜碱又与epitazettine 显然不同,所以推定它可能为B/D或C/D环系统结合不同于tazettine 的立体异构体。从红花石蒜L.radiata Herb.中分得7种已知生物碱:lycorine,homolycorine,tazettine,galanthamine,lycoramine,Iycorenine和pseudolycorine.从1种栽培的黄花石蒜中也分得上述7种生物碱。     

石蒜科生物碱的研究 Ⅶ.关于 suisenine

从中国水仙Narcissus tazetta L.var.chinensis Roem.的鳞茎中按木原报导的磷钨酸沉淀法分得其熔点[229℃,分解]与suisenine相同的物质,该物质经重结晶和纸上层析证明为假石蒜碱与石蒜碱的混合物,此外还分得tazettine.木原的suisenine样品经重结晶、纸上层析、电泳及混合熔点测定等证明同样为假石蒜碱与石蒜碱的混合物。     

石蒜科生物碱的研究 Ⅱ.假石蒜碱的新分离法和石蒜碱高含量植物的寻找

1.假石蒜碱用如下方法从6种石蒜属、5种水仙属植物中分得。卽用蓝氏铵盐使生物碱沉淀,溶于稀丙酮中,通过Zerolit 225(H型)。然后用4当量氨水洗脱该碱。另外我们也找到几种展开液,用它进行纸上层析,证明了微量的假石蒜碱。2.检查24种石蒜科植物,结果找到如下四种石蒜碱含量较高,卽:迟花水仙Leucojum vernum L.0.53%,雪铃花水仙Leucojum aestivum L.0.46%,矮喇叭水仙Narcissusincomparabilis Mill.0.33%,黄花石蒜Lycoris aurea Herb.0.20%。     

石蒜属植物忽地笑的生物碱类成分提取和结构鉴定

摘 要 目的:研究忽地笑生物碱部位的化学成分。方法: 采用各种色谱方法对生物碱部位进行分离,利用NMR、MS等波谱手段对所分离的生物碱成分进行结构鉴定。结果:分离得到12个生物碱：O-甲基石蒜伦碱 (1)、石蒜伦碱 (2)、O-乙基石蒜伦碱 (3)、力克拉敏 (4)、石蒜碱 (5)、加兰他敏 (6)、O-去甲基力克拉敏 (7)、N-去甲基力克拉敏 (8)、伪石蒜碱 (9)、漳州水仙碱 (10)、O-去甲加兰他敏 (11)、N-去甲基加兰他敏 (12)。结论:化合物3、8为首次从该植物中分离得到。     

人参四逆汤质量控制方法的研究——乌头类生物碱的鉴别、含量测定及乌头碱的限量检查

本文以人参四逆汤中附片的有效成分为指标。用薄层色谱法作乌头类生物碱鉴别及乌头碱的限量检查。用离子对萃取—分光光度法测定乌头类生物碱含量,乌头碱的回收率为98.33%±4.72%～103.6±3.09%,加样回收率为95.09±2.09%。方法准确、重现性好。     

槐果碱对HERG钾通道电生理功能的影响

HERG(human ether-a-go-go-related gene)钾通道在心律失常的发生及治疗中具有重要作用， 因此已成为近些年来的研究热点。本研究应用全细胞膜片钳技术记录在HEK(human embryonic kidney) 293细胞上稳定表达的HERG钾通道的电流和动力学曲线(激活、 失活、 复活和去活化)来研究不同浓度槐果碱对HERG电流及动力学曲线的影响， 以了解槐果碱抗心律失常的作用机制。结果表明， 槐果碱浓度依赖性地抑制HERG时间依赖性电流(I_step)及其尾电流(I_tail)。在0 mV时， 10、 30、 100及300 μmol·L^-1槐果碱对I_step的抑制率分别为(10.7±2.8)%、 (11.3±5.5)%、 (47.0±2.3)%及(53.7±2.5)%， 对I_tail的抑制率分别为(1.1±3.0)%、 (17.1±3.3)%、 (32.7±1.9)%(P<0.05， n=12)及(56.0±2.4)%(P<0.05， n=13)。100 μmol·L^-1槐果碱作用后失活时间常数减小， 失活速率变快；复活时间常数在大部分指令电压下明显减小(P<0.01， n=12)，复活速度加快；瞬时失活时间常数减小(P<0.05， n=12)；稳态激活、去活化无明显改变。由此可看出，槐果碱通过影响通道的失活过程抑制HERG钾电流，使得心肌细胞复极时间延长，改善快速性心律失常。     

槟榔加工法的试验

本文采用砂渍法软化槟榔,再刨成薄片,并与水浸泡法所得成品,进行试验比较,分别测定有效成分含醚溶性生物碱按槟榔碱计算,其结果:前法X±SD为0.3002±0.0021,损失率7.91%。后法X±SD为0.2762±0.0013,损失率15.27%,因此砂渍法对槟榔是一种行之有效的加工方法,值得推广。     

马钱子生物碱在大鼠体内的药代动力学研究马钱子生物碱在大鼠体内的药代动力学研究

目的用反相高效液相色谱法研究马钱子砂烫炮制品中生物碱在大鼠体内的药代动力学。方法HPLC法,色谱柱为Hypersil BDS C₁₈,流动相为乙腈-水,流速0.8 mL·min^-1,检测波长254 nm。结果大鼠iv本品后,其主要成分士的宁、马钱子碱、士的宁氮氧化物和马钱子碱氮氧化物的代谢均符合二室开放模型,主要药代动力学参数分别如下:T_1/2α为(8±5), (4±3), (6.2±1.7) 和(3.0±0.8) min,T_1/2β为(262±125), (416±131), (285±50)和(342±141) min,CL为(17±4), (21±12), (1.9±1.8)和(2.8±1.1) mL·min^-1,V_c为(1.4±0.5), (1.7±1.1), (0.24±0.16)和(0.23±0.06) L·kg^-1,V_d为(6.0±1.2), (12±7), (0.8±0.6)和(1.5±0.6) L·kg^-1,AUC为(57 578±25 578),(35 240±15 616),(93 088±22 375)和(177 712±120 110) h·μg·L^-1。结论此法为人体内的药代动力学研究提供参考。     

生物碱类浸膏制剂分析方法的研究 Ⅱ.番木鳖制剂的薄层色层分离——催化极谱测定

本文提出了应用氧化鋁薄层,分离番木鳖制剂中士的宁和马钱子碱的简单方法。薄层分离后,士的宁斑点用作者等改良的斑点收集管收集,以乙醇洗脫后于Mcllvaine缓冲液及0.1M碘化钾溶液的底液中测定其催化极谱电流。在浓度为3×10^-5—1.2×10^-4M范围內其浓度与催化电流值成一直线关系,可直接测定其含量。方法较为迅速简便,灵敏度较高且较为准确。最大测定误差为±2%。可用于番木鳖浸膏、番木鳖流浸膏及番木鳖酊的测定。     

反相高效液相色谱法测定骆驼蓬总碱片中去氢骆驼蓬碱和骆驼蓬碱的含量

目的 :建立RP HPLC法测定骆驼蓬总生物碱片中去氢骆驼蓬碱和骆驼蓬碱含量的方法。方法 :以甲醇 0 .0 1mol·L-1硫酸铵溶液 二乙铵 (6 0∶40∶0 .4,用磷酸调 pH至 (3.8± 0 .1)为流动相 ,进样量 2 0 μl,分别在 32 0 ,372nm处用RP HPLC法测定去氢骆驼蓬碱和骆驼蓬碱的含量。结果 :去氢骆驼蓬碱和骆驼蓬碱分别在浓度为 2～ 2 5 μg·ml-1的范围内与峰面积成良好的线性关系。方法的回收率分别为 (10 1.8± 1.8) %和 (10 1.3± 1.4) %。结论 :该法简便 ,准确 ,可靠     

Endothelium-dependent Relaxation in Response to Ethanol in the Porcine Isolated Pulmonary Artery

Many drugs cannot be dissolved in distilled water and so other solvents such as ethanol, dimethylsulphoxide and methanol are used. Because very little is known about the direct effects of these three solvents on the cardiovascular system, we have examined their effects on isolated pulmonary and coronary arteries from the pig. Increasing concentrations of ethanol, dimethylsulphoxide and methanol induced relaxation in porcine pulmonary (at 1·2% v/v, 59·9±9·0% (n = 9), 55·9±9·0% (n = 6) and 12·3±6·4% (n = 8), respectively, of U46619-induced tone) and coronary arteries (at 1·2% v/v, 69·9±7·1% (n = 10), 78·9±6·1% (n = 7) and 12·9±8·2% (n = 6) respectively, of U46619-induced tone). In the pulmonary arteries the relaxation in response to ethanol was found to be endothelium-dependent whereas the responses to dimethylsulphoxide and methanol were unaffected by removal of the endothelium. In the coronary arteries the relaxation to all three solvents was independent of the presence of the endothelium. Comparison of the sensitivity of the tissues to the solvents showed that ethanol and dimethylsulphoxide produced comparative responses in both the pulmonary and coronary arteries, whereas methanol was much less potent. The endothelium-dependent response to ethanol in the porcine pulmonary artery (maximum response, E_max, 67·1±9·3% of U46619-induced tone, n = 7) was attenuated by the cyclooxygenase inhibitor, flurbiprofen (E_max 31·9±12·0%, n = 7), the nitric oxide synthase inhibitor, L-NAME (N^G-nitro-L-arginine methyl ester; E_max 23·5±10·2%, n = 7)) and the combination of both inhibitors (E_max 18·3±7·8%, n = 7). The residual relaxatory response to ethanol was abolished, and converted into a contractile response, both by removal of the endothelium (at 1·7% v/v ethanol 27·3±11·5% of U46619-induced tone, n = 7) and by the addition of a low concentration of KCl (49·9±10·3%, n = 6), suggesting the release of a non-prostanoid, non-nitric oxide factor from the endothelium. This response, however, was not attenuated by the cannabinoid receptor-antagonist SR141716A (N-(piperidin-1-yl)-5-(4-chloro-phenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide HCl; 52·5±4·3% relaxation, n = 8), suggesting that the factor released in this preparation by ethanol is not a cannabinoid. The results of this study indicate that many solvents commonly used in pharmacological experiments have pronounced vasoactive properties. Methanol might be the vehicle of choice, because it was the least active solvent, whereas high concentrations of ethanol might influence vascular function at both the level of the smooth muscle and the endothelium, with the action on the endothelium involving the release of endothelium-derived relaxing factors.     

植物药中一些重要成分的测定方法的研究 Ⅱ.离子交换法在生物碱测定中的应用

离子交換剂用于植物药中生物碱的分析时,多为用溶剂提出后,通过离子交換树脂后进行測定。但1956年Brochmann-Hanssen提出将生药与离子交換树脂共同煮沸,則生物碱卽自生药中轉移到树脂上,可大大縮短操作时間,他曾經試驗了阿片、金鸡納(cinchona)、番木鱉(nux vomica)及吐根(ipecac)等生药,并与其他方法进行比較。但是这种方法并未引起广泛注意。文献中除这几篇以外并未再見报导。考虑到此法有快速簡便并且可以省去一般有机溶剂提取、节約原料等优点,我們也对一些植物药試用了这个     

氨硫脲的三种容量测定法的比较

关于氨硫脲即对乙酰氨基苯甲醛縮氨硫脲的含量测定法文献的报导是很多的。重量法的原理是根据氨硫脲与硝酸銀生成銀盐的作用。在容量法中有日本藥局方注解追补的酸鹼滴定法;以及硝酸銀法、溴量法、碘量法和三氯化铁法。很多比色法是采用酸或鹼使氨硫脲的乙酰基脫下而形成游离氨基,再經重氮化反应和偶合反应成为有色物質而比色的。此外,还有使氨硫脲与-溴代苯乙酮及与Grote氏試剂作用而比色的方法。     

合剂中山道年的极谱分析法

山道年及其制剂的分析法,有抽出后称重法,水解后滴定法,抽出后用2、4二硝基苯肼沉淀法,比色法等,此等方法,或操作麻煩,或所得結果很难准确,尤其是宝塔糖等赋料甚多的合剂,更無較好的方法。根据Santavy氏的报告,在含水酒精溶液中,可用極譜法测定山道年;在山道年、酚酞、糖、乳糖及可可的混合物中,用酒精抽取后,以硫酸鋰为支持电解質,可直接测定     

血液及尿中微量锑的比色测定

血液中微量锑的测定,目前仍广泛应用罗丹明 B 法.用此法测定微量锑时,为了免除铁离子的干扰作用,可利用醋酸戊酯在酸性溶液中将 HSbCl_6萃出,但必须控制溶液的酸度,当溶液酸度太高时,铁也同时被萃出.若利用结晶紫作为测定血液中微量锑的试剂,不但灵敏度大于罗丹明 B,空白试验着色浅,并且铁离子的干扰作用极其微小,因此在用醋酸戊酯提取 HSbCl_6时不必严格控制溶液的酸度.     

原儿茶醛分析方法的研究——Ⅱ.四季青和丹参中原儿茶醛的测定

应用间苯三酚和原儿茶醛在浓硫酸存在下的显色反应,结合薄层层析法建立了测定四季青和丹参中原儿茶醛的分析方法。本法简便准确,原儿茶醛在生药中的含量为万分之几而其测定结果的变更系数小于2%。     

反相高效液相色谱法测定普萘洛尔血药浓度及药代动力学的研究

This paper investigates the determination method of propranolol in human plasma by HPLC. YWG-C₁₈H₃₇ column was used with CH₃OH-H₂O-0.2mol/L KH₂PO₄ 0.2mol/L Na₂HPO₄(460:40:0.7:4.7)as mobile phaseThe preparation of plasma sample: Transfer 0.5 ml plasma into a 5 ml polystyrene test tube with cap, add 0.8 ml of 0.2% sodium hydroxide solution into the plasma. Propranolol was extracted with ether. External standard was used for quantitation.The assay was evaluated using the reproducibility within day and day-to day, and demonstrated high precision. Their average coefficients of variation were 5.8% and 6.8% respectively. The standard curve was linear (r=0.9999)over the range of 0.05～2.00μg/ml, the average recovery was 95.9±4.5%.     

Phytochemical investigation and radical scavenging activities of Melia azedarach and its DNA protective effect in cultured lymphocytes

Abstract

Context: Melia azedarach Linn (Meliaceae) is an Ayurvedic medicinal plant which is native to India. It is traditionally used for the treatment of leprosy, inflammation, scrofula, anthelmintic, antilithic, diuretic, deobstruent and cardiac disorders.

Objective: To evaluate the phytochemical constituents and antioxidant activities of the ethanol leaf extract of Melia azedarach (MA) and its protective effect against H₂O₂-induced cellular damage in cultured lymphocytes.

Materials and methods: The dose-dependent study of MA (20, 40, 60, 80, 100?µg/ml) was used to study in vitro radical scavenging assays. The effective dose of MA (60?µg/ml) was further used to study the H₂O₂-induced DNA damage (comet assay and DNA fragmentation assay) in cultured lymphocytes.

Results: The ethanol extract of MA (20, 40, 60, 80, 100?µg/ml) exhibited a significant dose-dependent inhibition of in vitro radical scavenging assays and their corresponding IC₅₀ values as follows: hydroxyl radical (26.50?±?0.26?µg/ml), superoxide anion (30.00?±?0.32?µg/ml), nitric oxide radical (48.00?±?0.48?µg/ml), DPPH radical (30.55?±?0.32?µg/ml) and reducing power (22.00?±?0.22?µg/ml). The increase in the severity of DNA damage and TBARS was increased significantly (p?<?0.05) at 500?µM H₂O₂-treated cultured lymphocytes and RBC cellular membranes. The phytochemical screening studies identified 13 chemical constituents present in the leaf extract of MA.

Discussion and conclusion: The results of this study demonstrate that MA offers protection against H₂O₂-induced cellular damage and it can be developed as an effective antioxidant during oxidative stress.     

Liquid extraction of ginseng and bilberry leaves

The number of diabetes patients continuously increases. Combined application of synthetic drugs and phytopreparations can decrease the negative side effects of drugs. The aim of this study was to create a new antidiabetic drug in the form of a liquid extract based on dry bilberry (Vaccinium myrtillus L.) and ginseng leaves. Quantification of flavonoids in bilberry leaves was carried out using a spectrophotometric method (425 nm); of tannins, by titration. Extraction performed with 50% aqueous ethanol isolated the maximum amount of principal active substances (polyphenols, ginsenosides). The optimum extraction conditions were determined as 4.5 h (time of infusion) and 5 h (time of displacement). Characteristics of the liquid extract are a brown fluid with distinctive odor, bittersweet, ethanol content 46.70 ± 0.31%, and solid residue content 35.23 ± 0.27%. The tannin content that was determined qualitatively using the reaction with FeNH₄(SO₄)₂ · 12H₂O (development of a dark-green color) and with 1% gelatin solution (formation of impurities) was 13.97 ± 0.22%. The flavonoid content that was determined by TLC (silica gel; butanol: acetic acid:water, 60:20:20; 3 spots of flavonoids) was 0.79 ± 0.03%. The panaxozide content that was determined by TLC (silica gel; butanol:ethanol:NH₄OH (25%), 9:2:5; 8 spots of panaxozides) was 0.29 ± 0.02%. __________ Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 42, No. 3, pp. 20–24, March, 2008.     

貉合滴定法在有机药物分析中的应用,II.士的宁盐类及其制剂的含量测定

士的宁盐类及其制剂的含量测定在各国药典中均采取提取生物硷后剩余酸量法或直接酸量法,但手续麻烦,费时多,取量大,且据Caws氏报告,士的宁与氯仿加热后能相互作用,生成结构不明的含氯反应物,妨碍结果的准确性。微量测定多用比色法,条件较难掌握。Budesinky氏曾报告士的宁的络合物滴定法,以Bi-EDTA-KI作为沉淀剂,游离出来之EDTA用硫酸锌滴定。