血清游离精氨酸的快速检测

建立快速、准确的精氨酸定量检测方法。采用 6 30 0黄金系统氨基酸分析仪 ,在锂柱生理体液分析方法基础上 ,建立血清游离精氨酸 (ARG)快速测定方法。血清样本经磺基水杨酸沉淀蛋白后取上清液进行色谱分析 ,色谱柱为Beckman公司阳离子交换柱 (12cm× 4 .0mm) ;流动相为 2 0mmol·L- 1 柠檬酸锂水溶液 ,流速为 2 0ml·h- 1 ;比色波长 5 70nm。该法检测精氨酸浓度的线性范围为 5mg·L- 1 ～ 5 0mg·L- 1 ,相关系数 0 .99834,最低检测限 1mg·L- 1 ,重复性 :日内RSD 0 .4 0 % ,日间RSD 0 .5 5 % ,回收率 97.6 7%～ 10 0 .6 7% (平均值 99.0 7% ) ;整个实验过程耗时 2 8min。该法简便、快速、准确、可靠 ,适用于临床和科研工作。     

梯度洗脱高效液相色谱法测定桑叶中3种活性成分的含量

建立同时测定桑叶中绿原酸、芦丁和槲皮素含量的分析方法。色谱柱为NUCLEODUR C18 RP(250 mm×4.6 mm,5μm),光电二极管检测器,流动相为甲醇-质量分数0.5%磷酸水溶液,梯度洗脱程序为0 min(30:70)-15min(30:70)-25min(50:50)-35min(85:15)-40min(30:70);流速0.8 ml.min-1,检测波长为350 nm。绿原酸、芦丁和槲皮素的线性范围分别为0.1144~1.0296μg(r=0.9996)、0.0436~0.3924μg(r=0.9999)和0.0452~0.4068μg(r=0.9997),平均回收率分别为97.7%(RSD=1.7%)、98.4%(RSD=2.2%)和100.6%(RSD=1.5%)。方法快速简便,专属性强,重复性好,可作为桑叶中绿原酸、芦丁和槲皮素的定量分析方法。     

氨基酸快速分析法测定高原低氧大鼠下丘脑γ-氨基丁酸的含量

建立一种快速、准确、可靠的γ 氨基丁酸定量检测方法 ,并观察高原低氧大鼠下丘脑γ 氨基丁酸 (GABA)含量变化。采用 6 30 0黄金系统氨基酸分析仪 ,在锂柱 130min程序生理体液分析方法基础上 ,根据γ 氨基丁酸 (GABA)的特性 ,建立了GABA的快速测定方法 ,并用此方法检测了高原低氧条件下大鼠下丘脑GABA的含量变化。结果高原低氧组大鼠下丘脑GABA的含量明显增多。此方法分析GABA的保留时间为 8.87min ,比原方法缩短了 6 5 .96min ;并且有较好的重现性 (CV :1.39% )、回收率高 (98.81% )。是一种快速、准确、可靠的GABA定量检测方法 ,可用于大批量样品的快速测定     

HPLC测定7种龙胆科植物花中龙胆苦苷与獐牙菜苦苷的含量

目的:建立7种龙胆花中龙胆苦苷和獐牙菜苦苷含量测定的HPLC方法。方法:采用微波辅助动态回流法进行提取,色谱条件:Fusion-RP 80 A C18柱(150 mm×4.6 mm,5μm);流动相:甲醇-0.2%磷酸溶液梯度洗脱(0~25 min:15%~30%);流速:1 mL/min;柱温:30℃;检测波长240 nm。结果:7种龙胆花中獐牙菜苦苷和龙胆苦苷的色谱峰与共存组分完全达到基线分离,线性范围分别为0.105~0.945μg(r=0.999 9),0.3~0.7μg(r=0.999 9),平均加样回收率分别为97.8%(RSD=1.02%),98.9%(RSD=1.51%)。结论:所建立的方法测定快速,结果准确可靠。     

高效液相色谱法检测发酵液中木糖和木糖醇

建立高效液相色谱检测发酵液中木糖和木糖醇含量的分析方法。色谱柱为HypersilNH2 柱 (4 .6mmi.d.× 2 5 0mm ,5 μm) ,柱温 3 5℃ ,流动相为乙腈—水 (80∶2 0 ) ,流速 1 .0mL .min 1,示差折光检测器检测。木糖和木糖醇在 3 .0～ 60mg.mL 1范围内 ,峰面积与其浓度线性关系良好 (г=0 .9995 ) ;平均回收率分别为 96.0 7% (n =5 ,RSD =0 .5 1 % )和 97.47% (n =5 ,RSD =1 .1 3 % )。方法简便、快速、准确。     

氨基酸分析法快速测定梭曼中毒后大鼠脑组织中兴奋性氨基酸的含量

建立一种快速、准确、可靠的脑内兴奋性氨基酸定量检测方法 ,并观察梭曼惊厥后大鼠脑组织中兴奋性氨基酸(EAAs)含量变化。采用 6 30 0黄金系统氨基酸分析仪 ,在锂柱 130min程序生理体液分析方法基础上 ,根据兴奋性氨基酸(EAAs)的特性 ,建立了EAAs的快速测定方法 ,并用此方法对梭曼惊厥后不同时相大鼠的新鲜脑组织进行定位检测。梭曼诱发惊厥后大脑皮质和海马内谷氨酸和天冬门氨酸水平显著下降。惊厥 30min时谷氨酸下降最明显 ,分别是正常组的 5 3.2 %和 5 2 .8%。天门冬氨酸更易受梭曼中毒的影响 ,惊厥后 5、30、90min 3个时相点测定值均显著下降。此方法完成谷氨酸和天门冬氨酸分析的时间是 2 0min ,比原方法缩短了 110min ;且有较好的重现性 (GluCV :日内 1.86 % ,日间 2 .32 % ;AspCV :日内 1.42 ,日间 2 .48% )和回收率 (Glu 97.7% ;Asp97.3% )。兴奋性氨基酸参与了梭曼中毒性惊厥的病理生理过程。本方法定量检测兴奋性氨基酸快速、准确 ,并利于大批量样品的快速测定     

高效液相色谱法直接测定当归中阿魏酸的含量

建立了一种高效液相色谱法直接测定当归中阿魏酸的含量。色谱柱为Shim-pack CLC-ODS(6.0 mm i.d×150 mm,5μm),流动相为甲醇—1.0%冰醋酸水溶液(体积比为25∶75),流速为1.0 mL/min,检测波长为313 nm。研究结果表明:阿魏酸的线性范围为1.0~10.0 mg/L,回归方程为A=13.352 8C 0.013 6(r=0.998 5),最小检测限为10 ng,加样平均回收率为94.67%,相对标准偏差(RSD)为1.57%。方法操作简便、快速、灵敏、准确。     

太子参及其种植土壤中16种元素的测定

目的:建立电感耦合等离子体原子发射光谱法(ICP-AES)测定太子参及其种植土壤中16种元素含量的方法.方法:采用微波消解处理样品,并考察了不同元素间相互干扰,采用ICP-AES同时测定16种元素.结果:对所测定元素的相时标准偏差(RSD)均小于4%,回收率在95%～104%之间,结果可信度高.结论:本研究所建立的方法具有简便、快速、准确、灵敏度高的优点.     

三种胆汁酸盐对Bilophila wadsworthia菌株生长的影响

本研究通过体外纯培养实验研究了3种胆汁酸盐:牛磺胆酸(TCA)、胆酸(CA)和脱氧胆酸(DCA),对沃氏嗜胆菌(Bilophila wadsworthia)菌株的生长状况的影响。比浊法和活菌计数法结果表明,TCA能显著促进该菌株生长,与添加相同摩尔浓度的牛磺酸(TAU)带来的效应接近,而CA和DCA对该菌株无显著作用。通过超高效液相色谱-质谱技术分析B.wadsworthia菌株对TCA的代谢的结果显示TCA可被该菌株分解产生CA和TAU。Ca2+沉淀法对B.wadsworthia菌株产胆盐水解酶的能力进行鉴定的结果表明该菌株具有产胆盐水解酶的能力。由此推断,TCA对B.wadsworthia菌株的促进作用主要是通过其水解产生的TAU,而非CA来发挥的。     

不同产地玛咖中酰胺含量分析

建立玛咖酰胺测定方法,测定不同产地玛咖切片中玛咖酰胺含量。采用HPLC法,色谱条件为Sepax Sapphire C18色谱柱(250 mm×4.6 mm,5μm)流动相:乙腈(A)-0.05%磷酸水溶液(B),梯度洗脱:0~15 min,90%~95%A;15~50 min,95%A。检测波长208 nm,体积流量1.0 m L/min,柱温30℃。酰胺1、酰胺2、酰胺3分别在0.88~132μg/m L、0.85~128μg/m L、0.96~144μg/m L线性关系良好。酰胺1、酰胺2平均回收率分别为99.21%(RSD=1.64%)、98.59%(RSD=1.75%)。不同产地玛咖中玛咖酰胺含量差别较大,建立可靠、准确、重复性好的检测方法,有利于评价不同产地玛咖的质量。     

Interaction between bile acids and staphylococcal polysaccharide: inhibition of capsule formation in encapsulated mutant strains (taurine+, taurine-) of Staphylococcus aureus

In a previous paper, we showed that bile acid derivatives inhibit capsule formation as well as taurine biosynthesis in a taurine+ (Tau+) encapsulated strain of Staphylococcus aureus. In the present study, binding of [14C]cholic acid [( 14C]CA) and [14C]taurocholic acid [( 14C]TA) to the staphylococcal polysaccharide antigen (SPA) of the capsular fraction was examined. The bile acids were found to bind with SPA via taurine of the Tau+ cells. [14C]CA bound with the SPA fraction of the Tau+ strain within 10-30 min, whereas 60-120 min was required in the binding of [14C]TA. Various bile acids competed with cholic acid binding to Tau+ cells which was shown by the inhibition of binding with cholic acid or taurocholic acid but not with glycholic acid. Binding of bile acid derivatives to a Tau- encapsulated mutant or to capsular material from this mutant was not observed.     

Xterra RP18柱高效液相色谱法快速分离测定氨基酸

建立了一种用XterraRP1 8色谱柱快速分离测定水解氨基酸的方法。所采用的色谱条件是 :WatersAlliance系统 ,柱温 5 6℃ ,流速 1 .8ml/min ,检测波长 2 4 8nm ,梯度分离 ,运行周期 2 5min,柱反压低于 2 0 0 0Psi。在 1 7.5min内分离了包括AMQ、NH3 和牛磺酸在内的 2 1种氨基化合物 ,适应于复合氨基酸注射液、含牛磺酸的氨基酸口服液及水解氨基酸样品的分析测定     

电渗析法进行胱氨酸母液脱盐的研究

猪毛酸解提取胱氨酸后的母液中含有十七种氨基酸。本文报道了采用我校由辐射法制备的高性能离子交换膜（ＨＦ－１及ＨＦ－２）,通过电渗析技术对母液进行脱盐并制得混合氨基酸。该技术已运转近一年,脱盐率＞９５．５％,氨基酸中人体必须氨基酸达２０％以上。     

Effect of taurine and gold nanoparticles on the morphological and molecular characteristics of muscle development during chicken embryogenesis

The objective of the present investigation was to evaluate the effects of taurine and Au nanoparticles on the expression of genes related to embryonic muscle development and on the morphological characteristics of muscles. Fertilised chicken eggs (n = 160) were randomly divided into four groups: without injection (Control) and with injection of Au nanoparticles (NanoAu), taurine (Tau) or Au nanoparticles with taurine (NanoAu + Tau). The experimental solutions were given in ovo, on the third day of incubation, by injecting 0.3 ml of the experimental solution into the air sack. The embryos were evaluated on the 20th day of incubation. The methods included gene expression at the mRNA and protein levels, immunohistochemistry, histology and microscopy. In groups NanoAu, Tau and NanoAu + Tau, the muscle structure and the number of muscle cells were affected. Furthermore, taurine increased fibre diameter, the total number of nuclei, the proportion of proliferating cell nuclear antigen (PCNA)-positive cells and the total cell number. Also, gene expression of basic fibroblast growth factor-2 and PCNA was downregulated. There were no significant interactions between NanoAu and taurine, indicating that NanoAu did not enhance the effects of taurine. It may be concluded that 20 days after injection, NanoAu affected some parameters of muscle development, but the most profound effects were those of taurine.     

Rapid and sensitive step gradient assays of glutamate, glycine, taurine and γ-aminobutyric acid by high-performance liquid chromatography–fluorescence detection with o-phthalaldehyde–mercaptoethanol derivatization with an emphasis on microdialysis samples

We developed a rapid step-gradient HPLC method for determination of glutamate, glycine and taurine, and a separate method for determination of γ-aminobutyric acid (GABA) in striatal microdialysates. The amino acids were pre-column derivatized with o-phthalaldehyde–2-mercaptoethanol by using an automated refrigerated autoinjector. Separation of the amino acids was established with a non-porous ODS-II HPLC column, late-eluting substances were washed out with a one-step low-pressure gradient. Concentrations of the amino acids were determined with a fixed-wavelength fluorescence detector. The detection limit for GABA was 80 fmol in a 15 μl sample, detection limits for glutamate, glycine and taurine were not determined because their concentrations in striatal perfusates were far above their detection limits. Total analysis time was less than 12 min, including the wash-out step. The methods described are relatively simple, sensitive, inexpensive, and fast enough to keep up with the microdialysis sampling.     

A rapid method for the determination of taurine in biological tissue

A rapid method for the determination of taurine is described. Based on high performance liquid chromatography (HPLC) separation, it avoids all transfers, precleaning treatments and the use of internal standards. The method consists of precolumn derivatization with O-phthaldialdehyde (OPA) followed by separation on a reversed phase styrene column using an acetonitrile-citrate buffer mixture as the eluting solvent. Naturally occurring substances, often quoted to co-migrate with taurine, are separated and the method described has been used to determine the taurine content of various tissues and blood fractions. These determinations indicate that the only satisfactory determination of blood taurine is that of whole blood.     

Stable isotope study of plasma taurine kinetics in rhesus monkey

Taurine is one of the most abundant amino acids in mammals and there is increasing evidence for the importance of taurine during development. Plasma taurine kinetics in a rhesus monkey was studied using [1,2-13C2]taurine. Taurine in plasma was derivatized to its dimethylaminomethylene methyl ester, separated on a gas chromatographic column, and the [M+2+H]+/[M+H]+ ion ratio was measured by ammonia chemical ionization mass spectrometry. The results were comparable to those obtained from the simultaneous radioisotope tracer study using [35S]taurine. This stable isotope method requires only 200 microliters of plasma for precise and accurate determination and is suitable for taurine kinetic studies in human infants.     

Quantitative determination of donepezil in human plasma by liquid chromatography/tandem mass spectrometry employing an automated liquid-liquid extraction based on 96-well format plates. Application to a bioequivalence study

An automated high-throughput liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was developed for quantitative determination of donepezil in human plasma. 150 MicroL of plasma samples were placed in 2.2 mL 96-deepwell plates and both donepezil and loratadine (IS) were extracted from human plasma by liquid-liquid extraction (LLE), using hexane as the organic solvent. Robotic liquid handling workstations were employed for all liquid transfer and solution preparation steps and resulted in a short sample preparation time. After vortexing, centrifugation and freezing, the supernatant organic solvent was evaporated and reconstituted in a small volume of reconstitution solution. The method developed, includes a sample analysis performed by reversed phase LC-MS/MS, with positive ion electrospray ionization, using multiple reaction monitoring (MRM). The chromatographic run time was set for 2.0 min with a flow rate of 0.7 mL/min in a C18 analytical column. The method was significantly sensitive, specific, accurate and precise for the determination of donepezil in human plasma and had the shortest run time. The curve was proven to be linear for the concentration range of 0.1-100 ng/mL. After validation, the method was applied to the rapid and reliable quantitative determination of donepezil in a bioequivalence study after per os administration of a 5mg donepezil tablet.     

Capillary electrophoretic separation and quantification of flavone-O- and C-glycosides in Achillea setacea W. et K

A simple method for the rapid separation and quantification of flavone-O- and C-glycosides in A. setacea W. et K. by capillary zone electrophoresis (CZE) with UV detection is described. Using 25 mM sodium borate with 20% (v/v) of methanol (pH 9.3) as running buffer sufficient separation of the analytes was achieved within 19 min. For the quantitative determination isorhamnetin-3-O-rutinoside was used as internal standard. The method was successfully applied to a rapid characterisation of the flavonoid complex and a precise quantification of the single and total amount of the flavonoids in different samples of A. setacea.     

Mutual Inhibition Kinetic Analysis of γ-Aminobutyric Acid, Taurine, and β-Alanine High-Affinity Transport into Neurons and Astrocytes: Evidence for Similarity Between the Taurine and β-Alanine Carriers in Both Cell Types

The transport kinetics of gamma-aminobutyric acid (GABA), taurine, and beta-alanine in addition to the mutual inhibition patterns of these compounds were investigated in cultures of neurons and astrocytes derived from mouse cerebral cortex. A high-affinity uptake system for each amino acid was demonstrated both in neurons (Km GABA = 24.9 +/- 1.7 microM; Km Tau = 20.0 +/- 3.3 microM; Km beta-Ala = 73.0 +/- 3.6 microM) and astrocytes (Km GABA = 31.4 +/- 2.9 microM, Km Tau = 24.7 +/- 1.3 microM; Km beta-Ala = 70.8 +/- 3.6 microM). The maximal uptake rates (Vmax) determined were such that, in neurons, Vmax GABA greater than Vmax beta-Ala = Vmax Tau, whereas in astrocytes, Vmax beta-Ala greater than Vmax Tau = Vmax GABA. Taurine was found to inhibit beta-alanine uptake into neurons and astrocytes in a competitive manner, with Ki values of 217 microM in neurons and 24 microM in astrocytes. beta-Alanine was shown to inhibit taurine uptake in neurons and astrocytes, also in a competitive manner, with Ki values of 72 microM in neurons and 71 microM in astrocytes. However, beta-alanine was found to be a weak noncompetitive inhibitor of neuronal and astrocytic GABA uptake, whereas in reverse experiments, GABA displayed weak noncompetitive inhibition of neuronal and astrocytic uptake of beta-alanine. Likewise, taurine was a weak noncompetitive inhibitor of GABA uptake in neurons and similarly, GABA was a weak noncompetitive inhibitor of taurine uptake into neurons.(ABSTRACT TRUNCATED AT 250 WORDS)