金银花中绿原酸的工业化提取纯化方法探讨

目的探讨金银花活性成分绿原酸的提取纯化工艺,使其能应用于工业化大生产。方法以绿原酸为检测指标,筛选最佳提取方法,考察比较四种树脂的纯化工艺,并对选定的D101大孔树脂富集纯化绿原酸的工艺条件进行考察。结果在所确定的条件下,经纯化后绿原酸纯度可达91.3%,绿原酸总转移率达55.7%。结论该提纯工艺对金银花有效成分的提取率高、稳定性好,可用于工业化生产。     

2-Chlorotrityl chloride resin

The esterification of 2-chlorotrityl chloride resin with Fmoc-amino acids in the presence of DIEA is studied under various conditions. High esterification yields are obtained using 0.6 equiv. Fmoc-amino acid/mmol resin in DCM or DCE, in 25 min, at room temperature. The reaction proceeds without by product formation even in the case of Fmoc-Asn and Fmoc-Gln. The quantitative and easy cleavage of amino acids and peptides from 2-chlorotrityl resin, by using AcOH/TFE/DCM mixtures, is accomplished within 15-60 min at room temperature, while t-butyl type protecting groups remain unaffected. Under these exceptionally mild conditions 2-chlorotrityl cations generated during the cleavage of amino acids and peptides from resin do not attack the nucleophilic side chains of Trp. Met, and Tyr.     

大孔吸附树脂分离纯化板蓝根有机酸组分

目的:采用大孔吸附树脂分离纯化板蓝根有机酸组分。方法:板蓝根以75%乙醇(pH2)索氏提取,以丁香酸为指标筛选吸附树脂,采用正交试验优化影响树脂吸附的因素,确定其合理工艺流程。结果:D101型树脂对板蓝根有机酸组分的吸附与解吸性能较好,采用2 BV(树脂床体积)50%乙醇易于洗脱;正交试验结果表明以药液浓度为1.0g.mL-1、pH3.0及流速为2 BV.h-1的参数组合应用效果良好,经济省时。结论:应用D101型大孔吸附树脂分离纯化板蓝根中有机酸组分,其综合性能良好,具有一定的应用前景。     

银杏叶提取物的树脂柱色谱脱酸工艺研究

目的 优化树脂柱色谱法脱除银杏叶提取物中总银杏酸的最佳工艺。方法 通过单因素和响应面法实验,以体积流量、洗脱剂乙醇体积分数、洗脱液接收体积为考察因素,以银杏叶提取物中总银杏酸去除率、总黄酮转移率和提取物收率的综合评分为指标,优化树脂柱色谱法脱除总银杏酸最佳工艺。结果 最佳工艺参数为体积流量1.2 BV/h,洗脱剂乙醇体积分数70%,洗脱液接收体积3 BV。结论 本工艺稳定、可行、重复性好,可作为银杏叶提取物树脂柱色谱脱酸工艺。     

大孔吸附树脂分离纯化大青叶有机酸部位的实验研究

目的:应用大孔吸附树脂分离纯化大青叶有机酸部位。方法:大青叶以70%乙醇(pH=2)索氏提取,以邻氨基苯甲酸为考察指标,优选最佳吸附树脂并采用正交试验优化影响树脂吸附的重要因素,确定其合理工艺流程。结果:以HPD100型树脂对大青叶中有机酸组分的吸附与解吸性能较好,采用3BV(树脂床体积)60%乙醇易于洗脱;正交试验结果表明以药液中邻氨基苯甲酸浓度为0.18mg.mL-1、pH3.5及洗脱流速为2BV.h-1的参数组合应用效果良好,经济省时。结论:应用HPD100型大孔吸附树脂分离纯化大青叶中有机酸活性部位,其综合性能良好,具有一定的药用前景。     

S-8型大孔吸附树脂纯化泽兰总黄酮(酚酸)的工艺研究

目的研究S-8型大孔树脂精制泽兰总黄酮（酚酸）的工艺条件。方法以总黄酮（酚酸）、咖啡酸含量结合指纹图谱主要特征峰面积保留率为考察指标;采用紫外-可见分光光度法测定总黄酮（酚酸）含量,HPLC测定咖啡酸含量并分析指纹图谱主要特征峰面积。结果 S-8型大孔吸附树脂纯化泽兰总黄酮（酚酸）的最佳工艺条件为：上样液浓度为含生药量0.1g·mL-1,上样量为每10g干树脂上样7g生药,吸附速率为1mL·min-1,水洗脱量为60mL,洗脱溶媒为80mL70%乙醇,洗脱速率为2mL·min-1;树脂经95%乙醇和1moL·L-1氢氧化钠溶液再生后,至少可重复使用5次。总黄酮（酚酸）与咖啡酸的保留率为75.56%及104.59%,峰1～5的保留率均达89%以上,纯化后总黄酮（酚酸）及咖啡酸的含量分别提高了3.99倍和4.69倍。结论 S-8型大孔吸附树脂能较好地纯化富集泽兰总黄酮（酚酸）。     

Resin glycosides from the leaves and stems of <Emphasis Type="Italic">Ipomoea digitata</Emphasis>

Alkaline hydrolysis of the ether-soluble resin glycoside (jalapin) fraction of the leaves and stems of Ipomoea digitata L. (Convolvulaceae) gave six organic acids, isobutyric, (S)-2-methylbutyric, tiglic, n-decanoic, n-dodecanoic, and cinnamic acids, and two glycosidic acids, quamoclinic acid A and operculinic acid A. Further, a new genuine resin glycoside, named digitatajalapin I, was isolated from the jalapin fraction, along with three known resin glycosides. Their structures have been determined on the basis of chemical and spectroscopic data.     

METHIONINE ENKEPHALIN AND ISOSTERIC ANALOGUES I. Synthesis on a Phenolic Resin Support†

An efficient synthesis of methionine enkephalin using a phenolic resin support is described. Analogues modified at their C-termini, such as peptide acids, amides, methyl esters and compounds formed by their reduction, were prepared conveniently from common peptide phenyl ester resins. The resin was used in the synthesis of complex isosterically modified analogues designed to investigate the role the peptide backbone plays in receptor interaction. Free hexapeptide phenyl ester resins underwent intramolecular aminolysis liberating the corresponding cyclic peptides.     

Synthesis of diketopiperazines with on‐resin N‐methylation and cyclative release

Abstract: Enantiomerically pure N‐methylated diketopiperazines (DKP) can be obtained by treating a N‐methylated resin‐bound dipeptide with 20% piperidine in dimethylformamide via a process known as cyclative release. N‐methylated resin‐bound dipeptides can be formed from N‐methylated precursors or N‐methylation can be selectively performed on the resin. When on‐resin N‐methylation was performed on the C‐terminal side of the dipeptide, diastereomers were formed. Yet the cyclative release is shown to be a stereoselective process, as seen using preformed N‐methylated amino acids. The procedure was also applied to synthesize the pseudodiketopiperazine cyclo(Pheψ[CH₂NH]Leu). When comparing nonmethylated, monomethylated and bismethylated derivatives, we find that N‐methylation results in a dramatic increase in solubility.     

3-乙酰基-11-酮-β-乳香酸的抗炎作斥及其衍生物的化学修饰

乳香酸（Bas）是乳香提取物中的活性成分,乳香酸在治疗炎症方面的疾病如类风湿关节炎、慢性支气管炎、哮喘以及慢性发炎性肠道疾病（溃疡性结肠炎和克罗恩病）已经显示出显著的药理活性。数据表明,3-乙酰基-11-酮-β-乳香酸（AKBA）被认为在各种乳香酸中是最强大的。AKBA被发现和确认为5-脂肪氧合酶（5-LOX）强大的抑制剂;AKBA的其他药理活性研究发现,它还充当了p38-MAP激酶强大的抑制剂。为了得到更多的AKBA,它的衍生物BA、KBA、ABA经过一系列化学修饰乙酰化和烯丙基氧化转化为AKBA,从而AKBA的量根据乳香树种和相应树脂质量从0．1％～3．0％上升到25％～35％。     

Boswellia serrata, a potential antiinflammatory agent: an overview

The resin of Boswellia species has been used as incense in religious and cultural ceremonies and in medicines since time immemorial. Boswellia serrata (Salai/Salai guggul), is a moderate to large sized branching tree of family Burseraceae (Genus Boswellia), grows in dry mountainous regions of India, Northern Africa and Middle East. Oleo gum-resin is tapped from the incision made on the trunk of the tree and is then stored in specially made bamboo basket for removal of oil content and getting the resin solidified. After processing, the gum-resin is then graded according to its flavour, colour, shape and size. In India, the States of Andhra Pradesh, Gujarat, Madhya Pradesh, Jharkhand and Chhattisgarh are the main source of Boswellia serrata. Regionally, it is also known by different names. The oleo gum-resins contain 30-60% resin, 5-10% essential oils, which are soluble in the organic solvents, and the rest is made up of polysaccharides. Gum-resin extracts of Boswellia serrata have been traditionally used in folk medicine for centuries to treat various chronic inflammatory diseases. The resinous part of Boswellia serrata possesses monoterpenes, diterpenes, triterpenes, tetracyclic triterpenic acids and four major pentacyclic triterpenic acids i.e. β-boswellic acid, acetyl-β-boswellic acid, 11-keto-β-boswellic acid and acetyl-11-keto-β-boswellic acid, responsible for inhibition of pro-inflammatory enzymes. Out of these four boswellic acids, acetyl-11-keto-β-boswellic acid is the most potent inhibitor of 5-lipoxygenase, an enzyme responsible for inflammation.     

The Chemical Shift Index method applied to resin‐bound peptides

Abstract: The Chemical Shift Index (CSI) method proposed by Wishart et al. [Biochemistry (1992) 31 , 1647–1651] to evaluate the secondary structure of peptides in aqueous solution uses as its reference the chemical shift values of each of the 20 natural amino acids (X) in a typical nonstructured sequence GGXAGG (17–20). In order to apply the CSI method to protected resin‐bound peptides, we established a new database of chemical shift values for the same GGXAGG sequences in their protected form and anchored to a polystyrene resin swollen in DMF‐d₇. The predictive value of this new reference set in the CSI protocol was tested on different resin‐bound peptides that were previously characterized by a full NOE analysis.     

THE SOLID PHASE SYNTHESIS OF PORCINE SECRETIN WITH FULL BIOLOGICAL ACTIVITY

The solid phase synthesis of porcine secretin is described. The C-terminal residue was attached to a polymeric amino support and all the Boc-amino acids, including Boc-glutamine, were coupled by a modified carbodiimide method. A preliminary test synthesis showed that the couplings of several amino acids of the N-terminal section were unsatisfactory. This problem was overcome in the main synthesis by executing all the major reactions twice. Cleavage of the peptide from the resin as well as the removal of all the side chain protecting groups was achieved with liquid HF. The product was purified by ion-exchange chromatography on SP-Sephadex to obtain a highly purified heptacosapeptide amide with full biological activity.     

胸腺五肽合成工艺的研究

采用固相合成法进行了胸腺五肽的合成研究,以Fmoc-Tyr(But)-Wang Resin为载体,所有氨基酸衍生物都采用Fmoc氨基酸保护系统,并以BOP-HOBT活化酯法偶联形成肽键,以20%哌啶/DMF为脱Fmoc试剂,以三氟乙酸(TFA)为脱氨基酸衍生物侧链保护基团和从树脂上切下肽试剂.经冷乙醚沉淀得粗品,以RP-HPLC纯化后得TFA型TP-5,经醋酸转型后冷冻干燥行TP-5白色粉末状物质.     

螺旋霉素发酵代谢物的分析及其对产物合成的影响

本文利用膜技术在螺旋霉素(SPM)发酵过程中让发酵液中的某些成份通过膜渗透到透析液中,然后用离子色谱分析仪、纸层析和氨基酸分析仪分析透析液中的代谢产物。经711阴离子交换树脂分离,发现洗脱液中含有乙酸、丙酸、丁酸、丙酮酸和草酰乙酸。用732强酸性阴离子树脂和纸层析初步分离和分析了透析液中天冬酰胺和谷氨酰胺的含量;经氨基酸分析仪测定,透析液中含有15种氨基酸。经静息细胞系统试验,发现短链脂肪酸有促进SPM合成的作用,而谷氨酰胺(1.0mmol/L)、天冬酰胺(2.0mmol/L)会抑制SPM的合成。膜过滤能降低发酵罐内谷氨酰胺和天冬酰胺浓度。这很可能是膜过滤发酵能提高SPM总产率的原因之一。     

Determination of chenodiol bioequivalence using an immobilized multi-enzyme bioluminescence technique

Measurement of the bioequivalence of formulations of chenodiol, a bile acid which is used for gallstone dissolution, is difficult because its high first-pass clearance results in low plasma levels after ingestion of usual dosages. To solve this problem, a new method was developed to determine the bioequivalence of several chenodiol formulations. The method included the following steps: isolation of all bile acids from serum by absorption to a hydrophobic resin, elution of bile acids from the resin by methanol, separation of the unconjugated bile acid fraction by an ion-exchange procedure, and bioluminescence measurement of the unconjugated 7 alpha-hydroxy bile acids using Sepharose beads containing co-immobilized 7 alpha-hydroxysteroid dehydrogenase, diaphorase, and luciferase. The isolation method gave complete recovery, and the bioluminescence procedure was simple, rapid, and sensitive. The peak level of systemic chenodiol occurred 1 to 2 h following oral ingestion and ranged from 4 to 8 microM. This method appears superior to previously reported methods for determining the bioequivalence of chenodiol preparations. In principle, the method is suitable for measurement of the bioequivalence of other bile acids provided the appropriate hydroxysteroid dehydrogenase is available.     

Components of convolvulin from <Emphasis Type="Italic">Quamoclit</Emphasis> × <Emphasis Type="Italic">multifida</Emphasis>

Alkaline hydrolysis of the ether-insoluble resin glycoside (convolvulin) fraction of the seeds of Quamoclit × multifida (syn. Q. sloteri House, Convolvulaceae), a hybrid between Q. pennat and Q. coccinea, gave three new glycosidic acids (maltifidinic acids C, D, and E) along with three known glycosidic acids (quamoclinic acids B, C, and D) and four organic acids (2S-methylbutyric, tiglic, 2R,3R-nilic, and 7S-hydroxydecanoic acids). The structures of the new glycosidic acids were characterized on the basis of spectroscopic data as well as chemical evidence.     

The tetrahydrocannabinol and tetrahydrocannabinolic acid content of cannabis products

Cannabinoid acids readily decarboxylate to the corresponding cannabinoid. Methods are available for the determination of Δ⁹-tetrahydrocannabinol (THC) and its acids (THCA) and published data on the levels of these compounds in cannabis are summarized. Using gas and liquid chromatography, fresh cannabis (64 samples) and cannabis resin (26 samples) from different countries were examined. Wide variations in the relative amounts of THCA and THC in cannabis were found. For cannabis resin, a wide range of values was also found (0·5: 1 to 6·1: 1), the lower values being in resins from the Indian sub-continent and the higher values in resins from the Mediterranean area. Total THC values were in the range 1·–10·6% in cannabis and 6·0–12·5% in cannabis resin.     

Two new glycosidic acids, multifidinic acids F and G, of the ether-insoluble resin glycoside (convolvulin) from the seeds of Quamoclit × multifida

Two new glycosidic acids, multifidinic acids F and G, were isolated from the glycosidic acid fraction afforded by alkaline hydrolysis of the ether-insoluble resin glycoside (convolvulin) fraction from the seeds of Quamoclit × multifida (syn. Q. sloteri House, Convolvulaceae), a hybrid between Q. pennata and Q. coccinea. The two compounds are the third and fourth examples of bisdesmosides of glycosidic acids having sugar linkages at C-3 of 3,11-dihydroxytetradecanoic acid (ipurolic acid) as well as at C-11.     

N-ALKYLATION OF AMINO ACID RESIDUES BY CHLOROMETHYL GROUPS

A serious side reaction in the peptide synthesis on the Merrifield resin was observed during attempts to synthesize the TMV fragment, Asn-Pro-Thr-Thr-Ala (101–105). The side reaction is consistent with N-alkylation of the amino groups by the residual chloromethyl groups on the resin, which lowers the total yield and complicates evaluation of monitoring data during the synthesis. It is shown that several amino acids can be N-alkylated in different positions in the peptide chain and in different solvents. The extent of N-alkylation is in some cases 50%.