壳聚糖树脂吸附提取辣椒素及其含量的测定

目的以干辣椒为原料,用壳聚糖树脂吸附提取辣椒碱并测定其含量。方法采用1%NaOH粗提取,提取液脱脂后用壳聚糖树脂柱吸附,乙醇洗脱、重结晶得淡黄色针状辣椒素的晶体。采用香兰素-亚硝酸钠比色法测定辣椒素含量,用HPLC分析晶体中辣椒碱和二氢辣椒碱的含量。结果辣椒素收率为0.38%,辣椒素晶体纯度为91.20%,其中辣椒碱为63.17%,二氢辣椒碱为26.75%。结论采用壳聚糖树脂吸附提取辣椒碱,方法简便,成本低廉,产率和纯度较高。     

硝酸银改性硅胶柱色谱法纯化川芎精油中的藁本内酯

目的采用硝酸银改性硅胶分离纯化中药川芎精油的活性化合物藁本内酯。方法传统硅胶层析柱中加入2％的硝酸银制备一种新型硝酸银改性硅胶层析柱,用石油醚-乙酸乙酯（98．5：1．5）进行洗脱,采用TLC和UV检测鉴别,将分离制得的藁本内酯用GC．MS进行结构鉴定,并用HPLC峰面积归一法进行纯度检查。结果从川芎精油中分离得到纯度为98．87％的藁本内酯,可以作为对照品使用。结论本研究在充分考虑化合物结构特性的基础上,建立了一种流程简单、成本低,藁本内酯纯度高的纯化工艺,可适用于现代化工业生产。     

三七皂苷K对照品的制备研究

目的 研究三七皂苷K对照品的制备。方法 利用大孔吸附树脂从三七剪口提取液中分离得三七总皂苷,再用反相柱色谱法分离得到三七皂苷K,并根据理化性质和波谱数据鉴定化合物的结构,同时运用薄层色谱（TLC）、高效液相色谱（HPLC）法检查及测定质量分数。结果 三七皂苷K单体经TLC检查无杂质斑点,二维及三维HPLC测定三七皂苷K的质量分数大于99%,峰质量分数因数为998.40。结果表明上述制得的纯品为单一化合物。结论 制备的三七皂苷K达到中药质量标准用化学对照品的技术要求。     

中药槐花饮片特征性提取物B的指纹图谱研究

目的：采用^1HNMR法和HPLC法研究中药槐花饮片特征性提取物B的指纹图谱，探讨炮制对槐花中特征性成分的影响。方法：按照规范工艺制备槐花、炒槐花和槐花炭的特征性提取物B（SCE B），分别测定各饮片的^1HNMR和HPLC指纹图谱。参照从槐花炭中分离得到的单体化合物（compound 1～5）的^1HNMR谱，分析3种饮片SCEB的^1HNMR谱，得到特征性成分的整体信息。以各单体成分为对照品分析3种饮片SCEB的HPLC指纹图谱，得到芳香类成分的信息。结果：3种饮片SCEB的^1HNMR谱基本一致，均显示主要为皂苷类成分的信息，还有少量芳香类成分的信息，表明炮制后其特征性成分未发生显著性变化。3种饮片的HPLC指纹图谱显示炒槐花与槐花的共有峰数目及各峰保留时间一致，而槐花炭中出现1个新的色谱峰，经对照品对照为化合物3，表明槐花制炭后有微量新成分产生。结论：中药槐花饮片的^1HNMR和HPLC指纹图谱分别反映了槐花炮制前后特征性成分的不同结构和整体组成，可作为其基源鉴定及炮制品鉴别的参考。     

高纬北极真菌Penicillium sp.C-5次级代谢产物的研究

目的对分离自北极Ny-lesund地区径流沉积物中的1株真菌Penicilliumsp.C-5的抗真菌活性成分进行研究。方法采用柱层析和半制备型HPLC等多种色谱方法对其发酵产物进行分离纯化; 运用MS、1 H-NMR和13 C-NMR等波谱方法,结合文献对照,确定化合物结构。结果从发酵液的乙酸乙酯萃取物中分得两个化合物,经鉴定为BE-31405和sordaricin。结论2个化合物均为首次从北极微生物中分离得到。     

一点红药材中异槲皮苷的分离鉴定及指纹图谱研究

目的:从一点红药材中分离黄酮类化合物异槲皮苷,同时建立一点红的HPLC指纹图谱分析方法。方法:采用半制备高效液相色谱法分离异槲皮苷,并以氢谱(1H-NMR)、碳谱(13C-NMR)、质谱(MS)确定其结构。采用高效液相色谱技术分析指纹图谱;色谱条件:苏州环球C18(250 mm×4.6 mm,5μm)色谱柱,乙腈-0.4%醋酸为流动相,梯度洗脱,流速1 mL·min-1,检测波长273 nm。结果:分离得到异槲皮苷,其纯度>98%;按照中国药典规定,11批一点红指纹图谱相似度均在0.9以上,确定了16个共有峰。结论:半制备高效液相色谱法制备异槲皮素方法简单、高效,指纹图谱分析方法准确性高、重复性较好,可作为一点红质量评价的依据。     

GC-MS方法分析肠激安方中的挥发性成分

目的采用GC—MS方法分析肠激安方中超临界CO2萃取物中挥发性成分。方法取萃取的挥发油用GC—MS法进行分析，结合计算机检索技术通过标准图谱对分离的化合物进行结构鉴定，应用色谱峰面积归一化法计算各成分的相对百分含量。结果超临界CO：萃取物挥发性成分各组分均得到较好分离，共分离出21个成分，鉴定了10个化合物。结论用GC—MS法测定肠激安方中超临界CO2萃取物挥发性成分的方法可行。     

洛伐他汀的杂质研究

目的从洛伐他汀结晶母液中分离纯化杂质成分,并进行结构鉴定。方法采用硅胶柱色谱、硅胶制备TLC、反相硅胶柱色谱、半制备HPLC等方法进行分离纯化;通过NMR、MS进行结构解析。结果分离得到6个单体化合物,分别鉴定为脱水Monacolin L,脱水二氢洛伐他汀,Monacolin L酸,Monacolin L酸甲酯,二氢洛伐他汀和二氢洛伐他汀酸。结论脱水Monacolin L为一新化合物。     

柱层析法分离精制辣椒红色素

目的制备高纯度的辣椒红素。方法采用乙醇渗漏法提取,硅胶柱层析法分离、石油醚重结晶精制,通过UV和MS对产品品质进行检测。结果得到深胭脂红色针状结晶(60~90℃的石油醚中结晶),mp:174~176℃。结论用柱层析法分离精制辣椒红素,具有操作简单,分离效果好,产品纯度高。     

高速逆流色谱法分离忍冬叶中木犀草素-7-O-β-D-葡萄糖苷和忍冬苷

利用高速逆流色谱法(HSCCC),从忍冬叶粗提物(35.2 mg)中一步分离制备木犀草素-7-O-?-D-葡萄糖苷(1,7.9 mg)和忍冬苷(2,5.6 mg)。HPLC峰面积归一化法计算得化合物1和2的纯度为98.0%和92.1%。     

Stability,permeation, and cytotoxicity reduction of capsicum extract nanoparticles loaded hydrogel containing wax gourd extract

The aim of this study was to develop hydrogel loaded with capsicum extract nanoparticles and wax gourd extract for transdermal delivery of capsaicin. The addition of wax gourd extract was supposed to reduce cytotoxicity of capsaicin in capsicum extract against HaCaT keratinocyte cell line. Capsicum extract nanoparticles were prepared by solvent displacement method using hyaluronic acid as a stabilizer. The physical and chemical stability of capsicum extract nanoparticles were investigated by dynamic light scattering technique and UV–Visible spectrophotometry, respectively. Hydrogel loaded with capsicum extract nanoparticles and wax gourd fruit extract was then formulated by using Carbopol 940® as a gelling agent for transdermal delivery. The skin permeability of capsaicin from the hydrogel was evaluated by Franz diffusion cell approach. The cytotoxicity reduction of capsicum extract nanoparticles and capsicum extract nanoparticles by mixing with wax gourd extract was determined by MTT assay The results showed that capsicum extract nanoparticles exhibited an average diameter of 168.4 ± 5.3 nm with a polydispersity index and zeta potential value of 0.26 ± 0.01 and −45.7 ± 7.1 mV, respectively. After two month-storage, particle size, polydispersity index, and zeta potential values of capsicum extract nanoparticles stored at 4° C, 30° C, and 45 °C did not significantly change. The capsaicin content decreased to 78%, 71%, and 72% when stored at 4 °C, 30 °C, and 45 °C for three months, respectively. The pH values of hydrogel containing capsicum extract nanoparticles were found to be in the range of 5.58–6.05 indicating good stability. The hydrogel exhibited a pseudoplastic character. The rate of permeation flux of capsaicin from hydrogel was 7.96 µg/cm²/h. A significant increase in cell viability was observed when the cells were incubated with capsicum extract nanoparticles mixed with wax gourd, compared to capsicum extract nanoparticles alone. The wax gourd extract in the hydrogel protected HaCaT cells from capsaicin cytotoxicity, thus may provide a new approach for delivery of capsaicin to reduce cytotoxicity to skin cells.     

Evaluation of oleoresin capsicum of Capsicum frutescenes var. Nagahari containing various percentages of capsaicinoids following inhalation as an active ingredient for tear gas munitions

Comparative efficacy as peripheral sensory irritant, oral and inhalation exposure studies were carried out on oleoresin capsicum (OC) of Capsicum frutescence var. Nagahari containing various percentages of capsaicinoids and two synthetic isomers of capsaicin in Swiss albino male mouse model to come up with a suitable active ingredient from natural source for tear gas munitions. The compounds screened were OC having varying percentages of capsaicinoids (20, 40 and 80%, respectively) and synthetic isomers (E and Z) of capsaicin (8-methyl-N-vanillyl-6-nonenamide). Mice were exposed to pyrotechnically generated smoke of the compounds in an all glass static exposure chamber for 15?min to determine acute inhalation toxicity (LC(50)) and quantitative sensory irritation potential (RD(50)). Acute oral median lethal dose (LD(50)) was also evaluated. Safety index of tear gas (SITG), a ratio of lethal concentration 50% (LC(50)) and the concentration which depresses respiration by 50% (RD(50)) due to peripheral sensory irritation is also proposed. The compound having highest SITG is considered as the most suitable to be used for tear gas munitions. The study revealed that oleoresin capsicum containing 40% capsaicinoids had the highest SITG among the compounds studied. The oral dosage versus mortality pattern of some compoundsdid not follow a true dose-response curve (DRC); however, following inhalation, all the compounds followed DRC. It was concluded that oleoresin capsicum (40% capsaicinoids) may be considered as the most suitable and environmental friendly compound from natural source to be used as an active ingredient for tear gas munitions.     

Interactions between A-9THC and capsaicin on isolated lamb bladder detrusor

A number of studies have demonstrated that capsaicin, a capsicum alkaloid, can affect isolated bladder tissue with either a relaxation or a contraction, depending on the species, by acting on VR1 receptors. In a previous work on isolated lamb detrusor, we demonstrated that capsaicin generally produces a relaxation of the tissue; this relaxation seems to be mediated by CGRP. Endogenous cannabinoids, such as anandamide, produce some of their actions by stimulating VR1 receptors and this seems to cause the release of peptides, e.g. CGRP. The aim of this work was to ascertain whether a cannabinoid, delta-9-tetrahydrocannabinol (delta-9THC), was able to interfere with the response of the isolated lamb detrusor to capsaicin. A-9THC, at concentrations between 1.6 x 10(-7) and 1.3 x 10(-6) M, displayed no activity on tissues. Instead, following delta-9THC, most of the tissues responded to capsaicin with a contraction that was abolished by atropine (9.0 x 10(-7) M). It has been reported that cannabinoids can inhibit the release of CGRP by stimulation of CB1 and CB2 cannabinoid receptors. Delta-9THC could act stimulating these receptors and thus inhibiting CGRP release and vesical relaxation. The muscle relaxing component removal could favour the contracting component, usually not active.     

复方双氯芬酸辣椒凝胶贴膏的质量控制及体外透皮

目的:建立复方双氯芬酸辣椒凝胶贴膏质量控制方法,并考察其体外透皮效果。方法:以HPLC法测定主药双氯芬酸二乙胺和辣椒素的含量,并测定凝胶膏剂含膏量、粘接性能、重量差异等指标;同时对其小鼠离体皮肤透皮特性进行了测试。结果:建立的HPLC法可同时测定双氯芬酸和辣椒素含量,双氯芬酸(以钠盐计)和辣椒素分别在25～800 mg.L-1和0.5～16.0mg.L-1内线性关系良好,平均加样回收率分别为98.49%和98.37%。小鼠透皮实验中,与市售扶他林乳胶剂相比,本凝胶膏剂中双氯芬酸二乙胺释放较慢,但24 h累积透皮量与扶他林近似;与市售好及施贴片相比,本凝胶贴膏中辣椒素释放较快和较多。结论:建立的方法可用于复方双氯芬酸辣椒凝胶贴膏的质量控制,其体外透皮效果良好,为开发凝胶膏剂新产品提供了实验依据。     

Evaluation of oleoresin capsicum of Capsicum frutescenes var. Nagahari containing various percentages of capsaicinoids following inhalation as an active ingredient for tear gas munitions

Comparative efficacy as peripheral sensory irritant, oral and inhalation exposure studies were carried out on oleoresin capsicum (OC) of Capsicum frutescence var. Nagahari containing various percentages of capsaicinoids and two synthetic isomers of capsaicin in Swiss albino male mouse model to come up with a suitable active ingredient from natural source for tear gas munitions. The compounds screened were OC having varying percentages of capsaicinoids (20, 40 and 80%, respectively) and synthetic isomers (E and Z) of capsaicin (8-methyl-N-vanillyl-6-nonenamide). Mice were exposed to pyrotechnically generated smoke of the compounds in an all glass static exposure chamber for 15?min to determine acute inhalation toxicity (LC₅₀) and quantitative sensory irritation potential (RD₅₀). Acute oral median lethal dose (LD₅₀) was also evaluated. Safety index of tear gas (SITG), a ratio of lethal concentration 50% (LC₅₀) and the concentration which depresses respiration by 50% (RD₅₀) due to peripheral sensory irritation is also proposed. The compound having highest SITG is considered as the most suitable to be used for tear gas munitions. The study revealed that oleoresin capsicum containing 40% capsaicinoids had the highest SITG among the compounds studied. The oral dosage versus mortality pattern of some compoundsdid not follow a true dose–response curve (DRC); however, following inhalation, all the compounds followed DRC. It was concluded that oleoresin capsicum (40% capsaicinoids) may be considered as the most suitable and environmental friendly compound from natural source to be used as an active ingredient for tear gas munitions.     

In vitro metabolism of R(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl]pyrrolo [1,2,3-de]1,4-benzoxazinyl]-(1-naphthalenyl) methanone mesylate, a cannabinoid receptor agonist.

R(+)-[2,3-Dihydro-5-methyl-3-[(morpholinyl)methyl]pyrrolo[1,2, 3-de]1,4-benzoxa zinyl]-(1-naphthalenyl methanone mesylate (WIN55212-2) is a potent cannabinoid receptor agonist that has been found to exhibit antinociceptive activity and to inhibit brain cyclooxygenase. The metabolism of WIN55212-2 has not been reported, and it is unknown whether its metabolites retain any agonist properties. In this study, in vitro metabolism of WIN55212-2 in rat liver microsome was investigated. The metabolic profile was obtained using high-performance liquid chromatography (HPLC) with UV and mass spectrometry detectors. The HPLC chromatogram revealed two major and at least six minor metabolites derived from the parent compound ([M + H](+) = m/z 427). The two major metabolites (structural isomers at m/z 461), constituting 60 to 75% of the total metabolites, were each identified as dihydrodiol metabolites resulting from the arene oxide pathway. The minor metabolites were all detected as protonated molecules, three of which appeared at m/z 477, corresponding to structural isomers of trihydroxylated parent compound; another two appeared at m/z 443, representing monohydroxylated isomers; and another was observed at m/z 425, and was assigned as a dehydrogenation product. These structural assignments are based on HPLC/tandem mass spectrometry and NMR analysis. Metabolic pathways have been proposed to account for the various metabolites observed. Two major metabolites have been isolated in pure form, allowing future receptor binding studies to be conducted.     

南极冰藻 Berkeleya rutilans H-15中清除自由基活性物质的分离纯化及其结构分析

目的 研究和发现极端环境下南极冰藻中清除自由基活性物质.方法 以南极冰藻Berkeleya rutilans H-15为试验材料,在国内外首先建立起了从南极冰藻中获取清除白由基活性物质的一套完整的方法,包括用甲畔提取活性化合物,用DPPH法和Godin法来追踪和检测活性化舍物的分离效果及变化情况,用硅胶柱层析对活性化舍物进行初步的分离和纯化,用Sephadex LH柱层析来进一步纯化,通过高效液相来进行化舍物纯度检测,高纯度化舍物用红外光谱和液质联用方法来确定化合物的类型和有关的功能基团.结果 在Berkeleya rutilans H-15中获得了一个纯的活性化舍物,经红外光谱和液质联用图谱初步进行分子结构分析,推测此化合物属于酚类化舍物,并具有很强的清除自由基的活性.结论 在南极冰藻中寻找并获得高活性的抗氧化剂是完全可行的.     

Capsicum pain plaster in chronic non-specific low back pain.

Topically applied capsaicin (CAS 404-86-4) induces the release of substance P, a neurotransmitter, from sensory C-fibres. In addition, there is a specific blockade of transport and de-novo synthesis of substance P. As a result, repeated applications of capsaicin bring about a long lasting desensitisation to pain (increase of pain threshold). The desensitising effect is fully reversible. The confirmed pharmacodynamic actions and a number of double-blind clinical studies indicate that local capsicum preparations are very suitable for the treatment of neuropathic pain or musculoskeletal disorders, with or without inflammatory components. In a double-blind, randomised parallel-group study a capsicum plaster was compared with a placebo for 3 weeks in 154 patients with non-specific back pain. Inclusion criteria were a history of back pain for a minimum period of 3 months and a degree of pain of 5 or more on an eleven grade visual analogue scale. The principal target variable consisted of the score of 3 combined pain scales. Secondary efficacy measures were tests of mobility, a disability index (in the context of Arhus low back rating scale) and global assessments by physicians and patients. For patients to be rated as responders their total pain score at the final examination after 3 weeks of treatment had to show a reduction by at least 30% of the baseline value. The study unequivocally achieved the target criterion with a rate of responders in the capsicum group of 60.8% against 42.1% in the placebo group (p = 0.0219). The sum of the 3 separate pain scales decreased more markedly in the capsicum group than in the placebo group (38.5% compared to 28.0%; p = 0.002). Relatively slight improvements of the impaired mobility and the functional status are explained by the characteristics of the disorder treated. The efficacy ratings by observers and patients was definitely in favour of capsicum. Adverse effects--mostly harmless and resolving spontaneously--were reported by 15 patients in the capsicum group and by 9 in the placebo group. The tolerance ratings by investigators and patients were superior to the placebo product. This, however, partly is due to the local pharmacological actions of the drug. As in comparably positive randomised studies with capsaicin cream in patients with osteoarthritis or fibromyalgia it was shown that a capsicum plaster preparation can also be used to advantage in chronic non-specific back pain.     

Capsaicin as an in vitro inhibitor of benzo(a)pyrene metabolism and its DNA binding in human and murine keratinocytes

Capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide), the active principle of capsicum fruits, such as hot peppers, is a known inhibitor of substance P. This substance was also found to be a potent in vitro inhibitor of human and murine epidermal metabolism of benzo(a)pyrene (BP) and the enzyme-mediated binding of BP metabolites to DNA. In both untreated and 3-methylcholanthrene-treated neonatal rat epidermal microsomes, capsaicin resulted in a dose-dependent inhibition of aryl hydrocarbon hydroxylase activity with an I50 value of 3.0 X 10(-4)-3.6 X 10(-4) M. A Lineweaver-Burk plot of the inhibition of aryl hydrocarbon hydroxylase activity suggested that the inhibition is of the noncompetitive type with Ki value of 50 microM. Capsaicin also inhibited BP metabolism and the binding of 3H-BP to DNA in BALB/c mouse and human keratinocytes. The formation of BP-7,8-diol was also substantially diminished in both systems in the presence of capsaicin (180-300 microM). Our results indicate that the substance P inhibitor, capsaicin, is also an inhibitor of epidermal BP metabolism and DNA binding of its metabolites. Therefore, in addition to its neurological effects, capsaicin may represent a new category of compound with anti-carcinogenic effects.