高效液相色谱法测定大豆乳清提取物中大豆异黄酮的含量

建立了大豆乳清提取物中大豆异黄酮含量的高效液相色谱测定方法。采用Nova-Pak C₁₈（3.9×150 mm,4 μm）色谱柱;以甲醇：0.4%磷酸=30：70（v/v）为流动相分析染料木苷和黄豆苷;流速为0.7 mL·min^-1;柱温为30℃;检测波长为260 nm。试验结果表明,大豆乳清提取物中的大豆异黄酮含量为72.5%,其组成以染料木苷和黄豆苷为主,二者比例接近1∶1,苷元型大豆异黄酮未检出。染料木苷和黄豆苷的平均回收率分别为98.1%和98.4%,相对标准偏差（RSD）分别为0.7%（n=5）和0.8%(n=5)。该方法快速、准确、重复性好。     

葛藤与葛根中异黄酮类成分的比较

分析比较了野葛〔Ｐｕｅｒａｒｉａｌｏｂａｔａ（Ｗｉｌｄ．）Ｏｈｗｉ〕的藤（葛藤）和根（葛根）的主要异黄酮类活性成分。从葛藤中首次分离得到３个化合物,经化学方法和光谱鉴定,证实为大豆甙元（Ａ）、大豆甙（Ｂ）和葛根素（Ｃ）。采用双波长薄层扫描法测定葛藤与葛根中上述３种异黄酮化合物的含量,葛藤中大豆甙元、大豆甙和葛根素的含量分别为０．１９５％,３．９３３％和２．４８１％;葛根中则分别为０．０５９％,０．７１４％和４．３１５％。研究结果为葛藤新药源的开发利用提供了科学依据     

超声提取和超声水解法从野葛根中提取纯化葛根素和大豆苷元

采用超声法从野葛根中提取葛根异黄酮,再将提取物在盐酸水溶液中超声水解结合有机溶剂萃取法从野葛根中分离纯化葛根素和大豆苷元。葛根素收得率为1．2％,纯度为97．8％;大豆苷元收得率为0．5％,纯度为98．2％。超声法从野葛根中提取分离葛根异黄酮活性成分葛根素和大豆苷元具有省时、节能、提取率和产品纯度高的优点。     

味噌异黄酮类化学成分

对味噌中异黄酮类化学成分进行了研究。通过硅胶柱色谱、Sephadex LH-20凝胶柱色谱、反相C18硅胶柱色谱以及制备HPLC等方法,对味噌正丁醇萃取部位进行分离纯化,根据化合物的理化性质和波谱数据(1H NMR、13C NMR、HMBC、MS)鉴定单体化合物的化学结构。总共分离得到6个异黄酮类化合物(图1),分别鉴定为:染料木素(1)、大豆素(2)、黄豆黄素(3)、4',5,7,8-四羟基异黄酮(4)、4',7,8-三羟基异黄酮(5)、4',7,8-三羟基-6-甲氧基异黄酮(6)。以上化合物均为首次从味噌中分离得到。     

野葛藤茎的异黄酮化学成分

从野葛Ｐｕｅｒａｒｉａ ｌｏｂａｔａ（Ｗｉｌｌｄ．）Ｏｈｗｉ藤茎中分离得到５个异黄酮类化合物,分别鉴定为：大豆甙元,芒柄花异黄酮,６,７－二甲３‘,４’－次甲二氧基异黄酮,大豆甙和葛根素。     

水提取法分离大豆异黄酮苷和苷元

本文建立了一种仅用水作为溶剂分离大豆异黄酮苷和苷元的方法。采用水加热提取的方法分离大豆异黄酮苷和苷元,分别对所用溶剂,提取温度,提取时间和物料比进行优化。实验上清液干燥后的固形物中大豆异黄酮苷含量为32．24％,苷元含量仅为3．05％,沉淀中大豆异黄酮苷元含量为72．03％,苷含量仅为4．28％。该方法经济,简单,绿色无毒,适合工业生产。     

铜藻岩藻黄素提取及纯化工艺研究

为了对岩藻黄素的提取、纯化进行系统研究,进而为高纯度岩藻黄素的工业化生产提供研究基础,筛选了适用于提取铜藻（Sargassum horneri）鲜藻中岩藻黄素的有机溶剂,并通过单因素实验和正交实验确定了最佳的提取溶剂浓度、提取温度、提取时间、料液比等工艺参数。随后采用硅胶柱层析法进行纯化,并通过单因素实验确定了最佳的硅胶柱床高度、上样量和洗脱流速。最后采用制备液相法对经层析纯化的岩藻黄素进一步纯化。结果表明,有机溶剂萃取的最佳工艺条件为：甲醇浓度90%,提取温度50 ℃,提取时间1 h,料液比1∶10,此条件下岩藻黄素提取率达到(0.258 9±0.003 6) mg·g-1鲜重（FW）［(1.078 8±0.015 0) mg·g-1干重（DW）］。硅胶柱层析的最佳工艺条件为：硅胶柱床高度10 cm,上样量6 g,洗脱流速10 mL·min-1,此条件下岩藻黄素得率为0.176 5 mg·g-1FW（0.735 3 mg·g-1 DW）,纯度为87.01%±0.88%。经制备液相进一步纯化后,岩藻黄素得率为0.127 1 mg·g-1 FW（0.529 4 mg·g-1 DW）,纯度为99.27%±0.22%。研究所用工艺简单,岩藻黄素得率高,为高纯度岩藻黄素的制备提供了实验基础。     

酸水解法从葛根中提取分离葛根素和大豆苷元

本文报道了葛根黄酮的提取精制方法,以６０％乙醇为溶剂,６０℃温浸６ｈ的优化工艺条件下,采用逆流萃取法从葛根中提取葛根黄酮,其收得率为１９．２８％,含量５１．０５％。葛根黄酮提取物采用５％盐酸水解４ｈ,酸水解液用乙酸乙酯萃取放置析出葛根素,乙酸乙酯相经水洗、浓缩和重结晶可得大豆苷元。采用酸水解法可以将葛根黄酮中的葛根素及大豆苷元衍生物水解成葛根素和大豆苷元,有利于葛根素、大豆苷元的分离及产率的提高;该方法从葛根中提取分离葛根素和大豆苷元具有操作简便、产品纯度和产率高、成本低的特点。葛根素和大豆苷元产率分别为１．３６％和０．４５％,纯度为９８．３２％和９１．２５％。     

光照对大豆幼苗组织中异黄酮含量和分布的影响

利用高效液相色谱（ＨＰＬＣ）测定了不同光照处理的大豆（Ｇｌｙｃｉｎｅｍａｘ（Ｌ．）Ｍｅｒｒｉ．）幼苗不同组织的异黄酮类含量。子叶中最高,叶片和根部相对较少。子叶的异黄酮以大豆甙和染料木甙及其丙二酰基结合体为主,且在光照条件下,异黄酮含量随光照时间的增加而显著升高;相反,黑暗中的异黄酮含量随苗龄的增加呈下降趋势;当子叶由黑暗转为光照处理以后,异黄酮含量同样随光照时间的增加而升高。在叶片和根部异黄酮含量和种类也因光照条件的不同而有很大差异。光照条件下,叶片中以染料木甙及其丙二酰结合体和黄酮芦丁为主,且随时间增加呈上升趋势;黑暗中的黄化叶片,则以大豆甙和丙二酰结合体为主,但随时间的变化不明显。在幼苗根部,黑暗条件下几乎检测不出异黄酮的存在;光照条件下,则可检测到５种异黄酮,其中以大豆甙元及其衍生物占主要部分。实验证实了光照对大豆异黄酮的积累有明显的促进作用     

高效液相色谱(HPLC)技术鉴定中国南方大豆品种异黄酮主要组分

采用高效液相色谱(HPLC)技术检测中国南方六个省份的249份大豆品种异黄酮主要组分含量.结果显示大豆籽粒中可检测出6种主要的异黄酮组分,分别为大豆甙(Daidzin)、甲氧基黄豆甙原(Glycitin)、染料木甙(Genistin)、丙二酰基大豆甙(Malonyldaidzin)、丙二酰基黄豆甙原(Malonylglycitin)和丙二酰基染料木甙(Malonylgenistin).各组分中以丙二酰基(Malonyl)异黄酮组分含量最高(61.2%),且各组分间相关极显著.大豆品种间异黄酮含量变异较大,变异系数达49.6%.来自江苏省的品种海门红黄豆乙异黄酮含量最高(4932.3μg/g),品种宝应等西风含量最低(367.1μg/g).不同省份间异黄酮含量差异极显著,来自浙江省的大豆品种平均含量最高(2717.2μg/g),来自安徽省的平均含量最低(1181.8μg/g).异黄酮含量与生育期呈极显著正相关(r=0.319* * *),与百粒重呈显著正相关(r=0.132*),而与脂肪含量(r=-0.45* * *)和蛋白质含量(r=-0.136)呈负相关.     

Acyl-coenzyme A: cholesterol acyltransferase assay: silica gel column separation of reaction products

Acyl-coenzyme A:cholesterol acyltransferase (ACAT) assays are usually performed by incubation of the enzyme with a labeled substrate followed by thin-layer chromatography separation and subsequent quantification of cholesteryl esters (CE) formed. Herein, a method is described for rapid separation of CE from other lipids, by elution from a silica gel column with a solvent mixture of petroleum ether/diethyl ether (98:2, v/v). Silica gel column chromatography is reliable and more rapid and safer than TLC. The best results were obtained when the reaction was stopped by Dole extraction followed by CE separation on a silica gel column. Assays for ACAT from rat intestinal microsomes showed that the specific activity values obtained using this method were reproducible and in good agreement with those obtained by conventional TLC method.     

Physiochemical properties of LIA-0191 A and B antibiotics

An antifungal antibiotic LIA-0191 was isolated from the mycelium by methanol extraction. It was shown with thin-layer chromatography that it consisted of components A and B. Component A was isolated with collumn chromatography on silica gel, recrystalization from the solvent mixture as a monocomponent crystalline substance. On the basis of the physicochemical and biological properties it was identified with sentacidin. Component B was obtained from preparation LIA-0191 by the method of counter-current distribution and recrystalization from methanol. Comparison of its physico-chemical and biological properties with those of the known purines and pyrimidine pyrrol showed that antibiotic LIA-0191 B is new.     

刺果紫玉盘的化学成分研究

采用95%乙醇提取,溶剂萃取,硅胶柱色谱分离等方法,从刺果紫玉盘根的乙醇提取物中分离得到了6个化合物,经NMR、MS等光谱学方法分别鉴定为:(-)1,6-desoxypipoxide(1)、piperenol A(2)、zeylena(3)、grandi-floracin(4)、顺式桂皮酸(5)和苯甲酸(6)。所有这些化合物都为从该植物中首次分离鉴定。     

Microquantitative determination of Pi-ATP and ADP-ATP exchange kinetics using thin-layer chromatography on silica gel

A new method for determination of 32Pi-ATP and [14C]ADP-ATP exchange rates is described. It is based upon separation of nucleotides and Pi by thin-layer chromatography on commercial aluminum or plastic sheets precoated with silica gel. The method permits avoiding special procedures for stopping of the reaction and for preparation of aliquots for thin-layer chromatography separation. It also allows to separate all adenine nucleotides and Pi in one chromatography procedure, and to work with a double label. The volume of the reaction mixture was 50-100 microliters. Aliquots (2-6 microliters) of the reaction mixture were taken at various moments without stopping the reaction and were layered immediately on a heated silica gel sheets. The nucleotides and Pi were separated in a solvent system which consisted of dioxane, isopropanol, 25% ammonia, and water (4:2:3:4, v/v). The nucleotide spots were detected in ultraviolet light, cut out, and their radioactivity was measured with a liquid scintillation counter. The method for measurement of kinetics of exchange reactions catalyzed by reconstituted into liposomes H+-ATPase complexes from beef heart mitochondria and high plant chloroplasts, is described.     

Isolation and preliminary characterization of carotenoids from pink-pigmented methylotrophs

An effective method was developed for complete removal of pigments from the cells and solvent mixture for further separation of pigments using thin layer chromatography on silica gel. Carotenoid samples that have been obtained in this way are of good purity for further investigations. Carotenoid pigments of pink-pigmented facultative methylotrophic bacteria Methylobacterium have been characterized. These carotenoids are represented mainly by xanthophylls, particularly hydroxycarotenoids. Strains M. fujisawaense B-3365 and M. mesophilicum B-3352 also have nonpolar carotenes in a small amount. Physico-chemical properties of carotenoids have been studied.     

A preparative method for purification of riboflavin 5'-monophosphate.

A preparative column chromatography method was developed for preparation of pure riboflavin 5′-monophosphate. A crude preparation of riboflavin phosphate(s) was chromatographed on DEAE-cellulose to provide a mixture of riboflavin 4′- and 5′-monophosphates. The 5′-isomer was isolated by chromatography on a column of silica gel with an ethanol:1 m triethylammonium bicarbonate, pH 7.5 (85:15) solvent system.     

Purification of growth-promoting peptides and proteins, and of histones, by high pressure silica gel chromatography.

A rapid method for the purification of histones and a variety of growth-promoting proteins and peptides by chromatography on silica gel has been developed. The isolation of the growth-promoting components of serum has been hampered by excessive losses associated with the use of water-based purification mens in acidic methanol-H2O solutions (eg. insulin, albumin, the somatomedins) provides a basis for purification on high-pressure silica gel columns, while peptides and histones can be purified in similar solvents. After column chromatography, the solvent is removed by flash-evaporation, or the protein may be precipitated directly from the solvent by neutralization of the pH and the addition of ethanol. The retention of biological activity (eg. somatomedin-C binding to insulin receptors and cell-growth stimulation) and recovery are excellent.     

Solvent systems for improved isolation and separation of territrems A and B.

Territrems A and B, tremorgenic mycotoxins in the rice culture of Aspergillus terreus, were extracted with hot chloroform. The toxins were cleaned on a silica gel column by direct adsorption-concentration of the extracts followed by desorption with chloroform-acetone (9:1, vol/vol). Crude toxin mixtures were separated by silica gel column chromatography and eluted with benzene-ethyl acetate (65:35, vol/vol). The method gave 112 mg of territrem A, 379 mg of territrem B, and 80 mg of their mixture from 4 kg of rice per batch. The criteria of extraction, cleanup, and separation are provided.     

Studies on neurosteroids: VII. Determination of pregnenolone and its 3-stearate in rat brains using high-performance liquid chromatography–atmospheric pressure chemical ionization mass spectrometry

An assay method for pregnenolone and its 3-stearate in rat brains has been developed using LC–atmospheric pressure chemical ionization (APCI) isotope dilution MS. The extraction of the rat brain homogenate containing deuterated internal standards (ISs) with organic solvent followed by silica gel mini-column chromatography gave two fractions containing pregnenolone and its 3-stearate together with the respective IS. Each fraction was derivatized into 3-acetate-20-methyloxime and 20-methyloxime, respectively, followed by silica gel mini-column chromatography to remove the excess reagents, and then each derivatized fraction was applied onto reversed-phase LC–APCI-MS. The method was applied to the determination of these steroids in the gray matter and olfactory bulbs of rat brains, which were divided into control and acute stressed specimens. Although pregnenolone in both regions of the rat brains increased more than five times after stress, its 3-stearate decreased after stress.