Regional Changes in the Cellular Level of Adenine Nucleotides in Ischemic Rat Brain Subjected to Single Embolization

Regional changes in adenine nucleotides in the rat brain were studied after 1 h of ischemia produced by the embolization method. The animals were divided into three groups according to neurological symptoms: sham-operation group, group A (hemiparesis only), and group B (hemiparesis with unconsciousness). Marked ATP depletion was detected in the hippocampus on the embolized side and extended to the other regions on the same side in group B. The results suggest that this damage in regional energy metabolism arises from regional reduction in blood flow and/or tissue vulnerability. ATP levels in the hypothalamus, hippocampus, and striatum on the opposite side of embolization decreased markedly in group B, and may be caused by extension of brain edema or diaschisis.     

高效液相色谱法同时测定荷叶中6种黄酮类成分

为了探索同时测定荷叶茶及饮片中6种黄酮类成分(芦丁、金丝桃苷、紫云英苷、槲皮素、山奈素和异鼠李素)含量的方法,本研究以8种不同的荷叶样本为材料,采用高效液相色谱分析法对6种黄酮类成分进行了同时测定。样品经过前处理,以0.5%甲酸-水(A)和0.1%甲酸-乙腈(B)作为流动相进行梯度洗脱,流速为1mL/min,柱温为25℃,进样量为20μL,经Agilent TC-C18(2)(150mm×4.6mm,5μm)色谱柱分离,于360nm波长处检测,结果显示,芦丁、金丝桃苷、紫云英苷、槲皮素、山奈酚、异鼠李素6种黄酮类成分分别在1.6~160、1.8~180、2.16~216、1.4~140、2.12~212、1.6~160μg/mL浓度范围内有良好的线性关系(R20.9992),其检测稳定性、重复性、日内精密度、日间精密度以及加样回收率的RSD均小于2%。进一步用该方法对不同来源的8个荷叶样本进行检测,结果显示荷叶样本中6种黄酮类成分含量以槲皮素最高,并且以样本G荷叶茶(购自G公司,批号为130802)的槲皮素含量最高。本研究建立的同时测定6种荷叶黄酮类成分含量的方法快速、准确,可为荷叶有效成分的检测和质量控制提供技术支撑。     

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%。     

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。     

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。     

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％。     

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。     

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。     

高效液相色谱法快速直接测定酪氨酸、苯丙氨酸和色氨酸

利用酪氨酸、苯丙氨酸和色氨酸具有紫外吸收这一特征 ,选用 2 3 0 nm、2 1 0 nm和 2 78nm的检测波长 ,在 7min内分别检测了酪氨酸、苯丙氨酸和色氨酸。三种氨基酸在 0 .0 1～ 1 .0 μmol/ ml浓度范围内呈显著的直线线性关系 ,相关系数均在 0 .9998以上。该方法不需衍生 ,直接测定 ,灵敏快速 ,结果准确可靠 ,适用于氨基酸注射液中低含量的酪氨酸和色氨酸准确检测 ,以及该三种氨基酸原料药纯度检测。     

A Quantitative Regional Analysis of Amino Acids Involved in Rat Brain Protein Synthesis by High Performance Liquid Chromatography

A biochemical method is described for the simultaneous quantitative estimation of unidirectional blood-brain amino acid influx and protein biosynthesis in individual structures of the rat brain. The method involved a double labeling experiment started by the administration of [14C]carboxyl-labeled amino acids and terminated 2 min after infusion of 3H-labeled amino acids, each at tracer quantities, the total labeling period being 45 min. Specific radioactivities of 14C- or 3H-labeled phenylalanine, tyrosine, leucine, isoleucine, and valine were determined in plasma and in small brain tissue samples for free amino acids, aminoacyl-tRNAs, and proteins. Amino acids were converted to their corresponding 5-dimethylamino-naphthalenesulfonyl (Dns, dansyl) derivatives and separated on HPLC C18 reversed-phase columns isocratically according to a newly developed optimizing procedure. The order of influx values between the neutral amino acids in relation to each other was Leu greater than Tyr greater than Ile greater than Phe greater than Val in every structure examined. Although aminoacylation of tRNAs was found to proceed to a comparable degree for neutral amino acids in all regions investigated, the specific radioactivity of amino acids attached to tRNAs differed substantially from that in the free amino acid pool, especially for leucine and valine. The results indicate the necessity of aminoacyl-tRNA determinations for tracer incorporation studies in protein synthesis analysis. Relative protein synthesis rates in the halothane-anesthetized rat were determined to be 30 and 67-91 pmol total amino acid incorporation/min/mg tissue for white and gray matter, respectively.     

Tyrosine Hydroxylase Activity in Rat Brain Synaptosomes: Direct Measurement Using High Performance Liquid Chromatography

Abstract: By use of high performance liquid chromatography with electrochemical detection to measure dopamine production, tyrosine hydroxylase (EC 1.14.16.2) activity has been measured in rat brain synaptosomes from striatum and forebrain. Normal specific activities three- to fivefold higher than previously reported in the literature for radiochemical methods of assay were found. It is suggested that synaptosomes contain a significant amount of endogenous substrate for tyrosine hydroxylase, which causes dilution of the added labelled tyrosine and hence underestimation of the activity of this enzyme when radiochemical methods are used.     

Energy Metabolic Changes in the Early Post-injury Period Following Traumatic Brain Injury in Rats

Impaired cerebral energy metabolism may be a major contributor to the secondary injury cascade that occurs following traumatic brain injury (TBI). To estimate the cortical energy metabolic state following mild and severe controlled cortical contusion (CCC) TBI in rats, ipsi-and contralateral cortical tissues were frozen in situ at 15 and 40 min post-injury and adenylate (ATP, ADP, AMP) levels were analyzed using high-performance liquid chromatography (HPLC) and the energy charge (EC) was calculated. At 15 min post-injury, mildly brain-injured animals showed a 43% decrease in cortical ATP levels and a 2.4-fold increase in AMP levels (P < 0.05), and there was a significant reduction of the ipsilateral cortical EC when compared to sham-injured animals (P < 0.05). At 40 min post-injury, the ipsilateral adenylate levels and EC had recovered to the values observed in the sham-injury group. In the severe CCC group, there was a 51% decrease in ipsilateral cortical ATP levels and a 5.3-fold increase in AMP levels with a significant reduction of cortical EC at 15 min post-injury (P < 0.05). At 40 min post-injury, a 2.6-fold ipsilateral increase in AMP levels and an 11% and 44% decrease in EC and ATP levels, respectively, remained (P < 0.05). A 37–38% reduction of the total adenylate pool was observed ipsilaterally in both CCC severity groups at the early time-point, and a 19% and 28% decrease remained in the mild and severe CCC groups, respectively, at 40 min post-injury. Significant contralateral ATP and EC changes were only observed in the severe CCC group at 40 min post-injury (P < 0.05). The energy-requiring secondary injury cascades that occur early post-injury do not challenge the brain tissue to the extent of ATP depletion and may provide a window of opportunity for therapeutic intervention.     

Phenylalanine Ammonia-Lyase Activity in Soybean Roots Extract Measured by Reverse-Phase High Performance Liquid Chromatography

Abstract: The high performance liquid chromatography (HPLC) technique was applied to measure phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) activity in soybean ( Glycine max L. Merril cv. BR16) roots. t -Cinnamate, the catalytic product of the PAL reaction was quantified at 275nm by isocratic elution with methanol: water through an ODS(M) column. Comparative experiments were carried out with 1.0 mM ferulic acid, an inducer of PAL activity. The results suggest that liquid chromatography is a rapid and sensitive method to analyze PAL activity in non-purified extract.     

柱前衍生HPLC法测定大鼠血浆中的河蚌糖胺聚糖

目的:建立柱前衍生高效液相色谱法(HPLC)用于河蚌糖胺聚糖在大鼠血浆中的含量测定。方法:使用异硫氰酸荧光素(FITC)对河蚌糖胺聚糖进行荧光标记,色谱柱为Shodex SB-804HQ(8.0×300 mm),流动相为流速为0.5 m L·min-1的0.1 mol·L-1的Na H2PO4-Na2HPO4缓冲液(p H 7.8),柱温30℃,激发波长495 nm,发射波长520 nm。结果:FITC对河蚌糖胺聚糖的荧光标记率为70;线性范围为5~120μg·m L-1,批内和批间精密度均小于15.0%,提取回收率为51.5~58.6%,样品在血浆中稳定性良好。结论:血浆中内源性杂质不干扰样品的测定,建立的方法符合生物样本的分析要求。     

ATP-levels of germinating seeds in relation to vigor

Determination of seed vigor was attempted by comparing ATP-levels of deteriorating seed to germination percentage and production of dry matter. Immediately after imbibition of any seed lot investigated, a production of ATP took place. This ATP-accumulation invariably reached a plateau after 6 h of imbibition. Two well germinating seed lots of rape, one of cauliflower and one of sugar beet, were artificially aged by means of elevated storage temperature and humidity. Every second week through 16 weeks of deterioration the levels of ATP, ADP and AMP after 7 h of imbibition were compared with the germination percentage. While ADP- and AMP-contents of germinating seed displayed no change (when imbibed 7 h) during the period of artificial aging, seed deterioration was reflected in the ATP-levels long before loss of viability could be detected by the conventional germination test.
When ATP-levels per seed were related to germination percentage throughout the aging, all four seed lots displayed similar patterns although the absolute figures differed. In contrast to the conventional "per seed' basis, however, ATP per gram seed not only displayed similar deterioration patterns, but the absolute values were also of the same magnitude.     

鸡肠中氨基酸含量的高效液相色谱分析

本文采用以异硫氰酸苯酯（ＰＩＴＣ）为柱前衍生化的高效液相色谱法测定氨基酸,并对方法测定条件进行了研究,得出流动相ｐＨ６．４０时测定的ＲＳＤ值最低,为０．５９％,样品经微波水解７ｍｉｎ（３００Ｗ功率）,且稀释液加入５ｍｉｎ后进样方能得到最佳结果。用本法对鸡肠中氨基酸进行了分析,必需氨基酸总量占３４．９％,具有可开发利用之价值。     

Content of Adenosine Phosphate Compounds in a Long-Day Duckweed, Lemna gibba G3, under Different Light and Nutritional Conditions

The levels of ATP ADP and AMP as well as the energy charge were examined in the long-day duckweed. Lemna gibba G3, under different light and nutritional conditions. ATP and ADP content, but not AMP, decreased slightly when the plant was cultured with a medium depleted of sucrose or was exposed to continuous darkness. The energy charge was not affected either by changes in the light conditions or by the depletion of sucrose from the medium, although increase of fresh weight changed drastically under these conditions. The levels of ADP and AMP and the value of the energy charge remained nearly constant throughout a 1-day period when the plant was exposed to short-day, continuous light and continuous dark. ATP content decreased gradually during the light period of the short day and thereafter remained constant during the dark period. When the plant was exposed to continuous light. ATP content decreased until the 8th hour after beginning of the continuous light, only to recover rapidly to its original level during the phase coinciding with the preceding dark period of the short day. Under continuous dark conditions the ATP content remained constant throughout the day. It was concluded that the diurnal rhythm of physiological activities previously reported cannot be related to the energy charge.