The Isomeric Composition of Hydroperoxides Formed by Autoxidation of Unsaturated Triglycerides and Vegetable Oils

Trioleoylglycerol (TO), trilinoleoylglycerol (TL), and trilinolenoylglycerol (TLN)were autoxi-dized in the dark at 37°C. Monohydroperoxides (MHP), the primary products, were isolated by preparative thin-layer chromatography (TLC). The isomeric compositions of their hydroperoxy fatty acid components were determined by gas chromatography-mass spectrometry (GC-MS) as follows—TO: the 8-, 9-, 10-, and 11-isomers; TL: the 9-, and 13-isomers; and TLN: the 9-, 12-, 13-, and 16-isomers. The proportions of isomers in each MHP did not vary with the oxidation time. The isomeric compositions of hydroperoxy fatty acid components obtained from autoxidized soybean and olive oils indicated that each unsaturated fatty acyl group of triacylglycerol (TG) in vegetable oils produced isomeric hydroperoxides during autoxidation in a way similar to the corresponding fatty acid methyl esters. The proportions of the isomers obtained from autoxidized oils changed with the level of oxidation. Isomers coming from linolenic acid in soybean oil and those from linoleic acid in olive oil decreased remarkably at a high level of oxidation.     

高效液相色谱法测定辣椒素

用95%醋酸钠饱和乙醇溶液抽提辣椒树脂油中的辣椒素,经高效液相色谱(HPLC)分离,在紫外波长280 nm处测定。用此法分析测得福建特产"小米椒"的辣椒素含量为10.38%。     

Separation of Glycopeptides by High Performance Liquid Chromatography

A method is presented for separation of tryptic glycopeptides-containing oligosaccharides of the N-asparagine-linked type. High performance liquid Chromatography (HPLC) of glycopeptides on a C18 reverse-phase system eluted with a gradient of 0%–50% acetonitrile in 0.1 M NaPO₄ pH 2.2 resolves the two major glycosylation sites from the envelope glycoprotein (G) of vesicular stomatitis virus. Glycopeptides containing N-linked oligosaccharides of the complex type coelute with those containing N-linked oligosaccharides of the neutral, high mannose type, indicating that separation is based upon peptide rather than carbohydrate composition. The contribution of the carbohydrate component to glycopeptide elution, as determined by cleavage of the high mannose oligosaccharides with endo-β-Nacetylglucosaminidase H, is that of a significant, but minor, decrease in peptide retention time. Comparison of the tryptic glycopeptide profiles of G isolated from both wild type and mutant strains of VSV illustrates the rapid, reproducible, and quantitative nature of the technique. Through HPLC analysis of appropriately treated glycopeptides, it is possible to explore both the nature and extent of glycosylation at individual sites in glycoproteins in a single step.     

Capsaicinol: Synthesis by Allylic Oxidation and Its Effect on TRPV1-Expressing Cells and Adrenaline Secretion in Rats

Capsaicinol is an ingredient of hot red pepper. In this study, we developed a novel method for capsaicinol synthesis and examined capsaicinol’s physiological effects on capsaicin receptor (TRPV1)-related actions. Allylic oxidation of capsaicin by palladium acetate (Pd(OAc)₂) resulted in the formation of (±)-capsaicinol acetate at a 7.2% yield in a single step. The effectiveness of (±)-capsaicinol in TRPV1 activation (EC₅₀=1.1 μM) was found to be weaker than that of capsaicin (EC₅₀=0.017 μM), whereas the efficacy of (±)-capsaicinol reached 75% of that of capsaicin. Intravenous administration of (±)-capsaicinol in anesthetized rats dose-dependently enhanced adrenaline secretion from the adrenal gland. The response to a 5 mg/kg-dose of (±)-capsaicinol was comparable to that of a 0.05 mg/kg-dose of capsaicin. The relative pungency of capsaicinol to capsaicin was coincident with the relative effectiveness in inducing these TRPV1-related actions.     

高效液相色谱法测定超滤后血浆中的谷氨酰胺

用高效液相色谱法测定超滤后血浆中的谷氨酰胺, 平均回收率为96.75%, 在130～1300μmol/L范围内呈线性关系(r=0.9906), 健康人正常值男性为(694.97±102.31)μmol/L, 女性为(623.79±99.27)μmol/L, 男女间值有显著差别(P＜0.05)该分析方法简单、迅速, 可用于外科肠道内营养的监测和对肠粘膜结构与功能的研究.     

制备型高效液相色谱法分离蛋白质的研究

生物工程的发展,特别是生化制品下游处理技术的兴起,对现代分离科学提出了更高的要求,研究和开发各类生化物质特别是活性生物大分子的分离纯化技术,已成为一项十分重要的研究课题。在现有的分离技术中,液相色谱,尤其是80年代在分析型高效液相色谱(HPLC)基础上兴起的制备型HPLC,在大规模分离纯化生物活性物质特别是蛋白质方面已显示出巨大的应用潜力,引起了各国研究者的高度重视^[1-5].本文利用自行设计的制备型HPLC分离装置,对牛血清白蛋白(BAS)和牛血清红蛋白(HG)的制备分离过程进行了实验研究,着重考虑了流动流速、柱超载方式、柱长等因素对BAS和HG分离度的影响。     

高效液相色谱测定血清中的非胆固醇甾醇

本文介绍一种用高效波相色谱（ＨＰＬＣ）同时测定血清中２４－脱氢胆固醇、７－烯胆甾烷醇、菜油甾醇、胆甾烷醇和β－谷甾醇的方法。血清加内标（６－氯豆甾醇）后用氢氧化钾醇溶液皂化,用正己烷提取其中的各种甾醇,将提出的甾醇衍生为苯氨基甲酸酯,用ＨＰＬＣ分离测定。本法样品处理简单,色谱分析时间短,批内和批间变异系数分别为２．２～３．８％和３．３～６．７％。本文也首次报告我国成年人血清非胆固醇甾醇水平,５种甾醇血清浓度的总和平均为１．５４ｍｇ／ｄｌ,是胆固醇的０．８６％。     

高效液相色谱法测定血浆中同型半胱氨酸

建立了一种高效液相色谱-库仑阵列电化学测定血浆中同型半胱氨酸的方法。采用3-2-羧乙基膦作为还原剂还原巯基间形成的二硫键,用0.1 mol.L-1HC lO4来沉淀血浆蛋白,离心取上清,进样分析。在0.40~50.00μmol.L-1范围内,线性关系良好。加标平均回收率是98.43%,RSD%是4.35%。该方法简单、快速、灵敏。     

Preparation of Porcine Neurophysin Proteins by High Performance Liquid Chromatography

Abstract: The crude neurophysin containing extract from posterior lobes of porcine pituitaries was roughly purified by gel chromatography. 15 mg of the lyophilized neurophysin complex were completely separated by HPLC yielding in neurophysin I₁ (3.6 mg), I₂ (4.0 mg), II (4.6 mg) and III (1.9 mg). All of the neurophysins were homogenous by PAGE and SDS-electrophoresis, isoelectrofocussing, amino-acid composition and N- and C-terminal amino acid analysis. In conclusion, HPLC is a reliable and quick method for the preparation of pure neurophysins.     

高效液相色谱测定微量胆固醇氧化产物

介绍一种高效液相色谱测定胆固醇氧化产物的方法,以苯甲酰氯为衍生剂,将胆固醇及其氧化产物衍生成苯甲酸酯,反相高效液相色谱分离,紫外检测,内标法定量．本法灵敏度高,重现性好,可应用于胆固醇纯度标准物质的杂质分析及血清中的胆固醇氧化产物测定。     

Characterization of Alkylgalactolipids from Calf Brain by High Performance Liquid Chromatography

Minor nonpolar galactolipids were isolated from the total lipids of calf brain stem by column chromatography and were separated by preparative thin-layer chromatography into four groups. The material recovered from the bottom band of the thin-layer chromatography consisted of monogalactosyl diglyceride and its 1-0-alkyl isomer, alkylgalactolipid, present in a molar ratio of 11 :9. After perbenzoylation. they were separated by preparative thin-layer chromatography and characterized. The fatty acid compositions of these lipids were similar to each other and to those of the ester-linked fatty acids of cerebroside esters. The major alkyl group of alkylgalactolipid was palmityl, and the other, minor components were oleyl. myristyl, and stearyl ethers. Perbenzoylated derivatives of these lipids were further separated by reverse-phase high performance liquid chromatography. The chromatograms from these two lipids were similar; however, most of the peaks were still mixtures of homologs containing different fatty acids or an alkyl group.     

高效液相色谱法同时测定17种磺胺类药物的研究

建立了高效液相色谱-紫外法同时测定17种磺胺类药物的分析方法.主要研究了色谱柱、流动相配比对磺胺类药物分离的影响.通过研究,确定了最佳液相色谱分析条件.分离条件为:YMC ODS-C18柱;以2%乙酸溶液、乙腈和甲醇作为流动相,进行梯度洗脱;检测波长为270 nm.该方法的检测限为:磺胺胍和磺胺为2.0 ng/ml,磺胺二甲氧哒嗪、磺胺二甲基嘧啶、磺胺多辛、磺胺胍、磺胺甲基嘧啶、磺胺甲噻二唑、磺胺甲基异噁唑、磺胺甲氧哒嗪、磺胺6-甲氧嘧啶、磺胺二甲基噁唑、磺胺、磺胺吡啶、磺胺喹噁啉、磺胺噻唑和磺胺异噁唑均为5.0 ng/ml.各组分的回收率在81.3%～97.9%,相对标准偏差在0.1%～4.3%.     

Separation of Lepidopterous Sex Pheromones by Reversed-phase Thin Layer Chromatography and High Performance Liquid Chromatography

Bis(4-chloro-2-ethylphenyl) phenylphosphonate was metabolically transformed into the cor-responding cyclic ester, i.e., 6-chloro-4-methyl-2-phenyl-4/f-1,3,2-benzodioxaphosphorin 2-oxide, in houseflies in vivo. In a p-unsubstituted analog, hydroxylation at the para-position of an ester linkage occurred preferably to alpha-hydroxylation with subsequent cyclization. The cyclization was diastereomerically selective, giving predominantly the cis ester. The biological activities of synthesized and related cyclic esters were similar to but weaker than saligenin cyclic phosphorus esters lacking a methyl group at the 4-position.     

高效液相色谱法测定山栀子中栀子甙的含量

采用高效液相色谱法测定中药山栀子（Cardenia jasminoides Ellis）中保肝利胆的主要成分栀子甙（geniposide）的含量。分析结果表明,本方法重现性好,平均加样回收率为100．90％,RSD为2．00％。用此方法测定未知样品山栀子中栀子甙的平均含量为3．463g/100g。     

Measurement of Aldehydes in Low Density Lipoprotein by High Performance Liquid Chromatography

A modified procedure is presented for the HPLC determination of nanomolar concentrations of n-alkanals, hydroxyalkenals, malondialdehyde and furfural in biological fluid. The modifications allow aldehyde profile analysis of small samples of fresh, human, low density lipoprotein (LDL), enabling more detailed studies of LDL fatty acid peroxidation. Aldehydes are reacted with 1,3-cyclohexanedione to produce fluorescent derivatives which are separated by gradient, reversed phase, high performance liquid chromatography (HPLC). Analysis time has been reduced by shortening the sample preparation. Sensitivity has been increased by miniaturization of the derivatisation procedure, reducing required sample size. Recoveries of added aldehydes have been improved. In addition, the method presented allows determination of three further aldehydes, not measured previously by CHD methods: malondialdehyde, formaldehyde and furfural. Recovery and variability data and concentrations of aldehydes found in human LDL are given. The capacity of the method for further development, to enable determination of other aldehydes such as the trans, 2-alkenals, is also demonstrated.     

Separation of Fuchsin Homologs Using High Performance Liquid Chromatography

Commercial samples of basic fuchsin contain variable proportions of four homologs, pararosaniline, rosaniline, magenta II, and new fuchsin. That different samples of dyes give variable staining is documented in the literature. Three commercial samples of basic fuchsin were investigated using high performance liquid chromatography. Separation of homologs was achieved using a C18 adsorbant and a solvent system of methanol:water:glacial acetic acid (66:24:10).     

Purification of Human Placental Trophoblast Interferon by Two-Dimensional High Performance Liquid Chromatography

Abstract

Human placental trophoblast challenged with Sendai virus induced IFNs mainly of the β-type (75%) and relatively low levels of the α-type (25%). A two-step high performance liquid chromatographic procedure (“two-dimensional HPLC”) has been developed for the complete purification of the placental trophoblast interferon beta (tro-IFN-β) from serum-containing culture supernatant. The method involved a combination of high performance liquid affinity chromatography (HPLAC) on Cibacron Blue 3GA immobilized on an activated pressure stable macroporous synthetic polymer, 2-hydroxyethyl methacrylate vinyl sulphone (HEMA-BIO 1000 VS), as the first dimension and reversed-phase high performance liquid chromatography (RP-HPLC) on Separon SGX C-18 as the second. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot experiments showed that the tro-IFN-β was present as a 24 kDa protein. Densitometric scanning analysis of Coomassie-stained gel revealed the purity of the final preparation to be greater than 99%. The purified tro-IFN-β had a specific activity of 1.03 × 10⁸ IU/mg of protein and the overall recovery was 81% of the total IFN-β activity in the crude preparation and 61% of the total IFN activity.