非转基因和转基因小麦中氨基酸含量的分析及比较

目的：对小麦中氨基酸分析的水解方法进行研究,比较非转基因和转基因小麦中氨基酸的含量。方法：样品用含0.1%巯基乙酸的盐酸溶液水解提取,样液经定容、过滤、浓缩、复溶、过膜后,用氨基酸分析仪测定,外标法定量。结果：应用含巯基乙酸的盐酸水解液对小麦样品进行水解的同时有效防止了含硫氨基酸（蛋氨酸和半胱氨酸）的氧化,相对氧化水解方法节省前处理时间,方法的回收率为93%~99%,相对标准偏差小于2.4%。非转基因和转基因小麦样品的17种氨基酸分析,氨基酸含量不存在显著性差异。结论：该方法准确快速、重现性好,适用于小麦中氨基酸含量分析的检测要求。     

云南盈江县不同产地草果氨基酸和矿质营养元素分析

用氨基酸自动分析仪、等离子体发射光谱仪(ICP法)等方法测定云南盈江县三个草果Amomum tsaoko主产乡的草果氨基酸、矿质营养元素等成分。结果显示,不同产地草果中氨基酸和矿质营养元素含量有明显差异。盈江县各地的草果氨基酸含量均比文山产草果低,其中苏典乡劈石村的2号样品(海拔1540 m,土壤pH 6.58)氨基酸含量与文山样品最为接近;微量营养元素总含量则相反,盈江县产草果均高于文山样品。     

牛乳基和羊乳基奶粉中16种氨基酸含量的测定及比较

目的：对乳粉中氨基酸分析的水解方法进行研究,比较牛乳基奶粉和羊乳基奶粉中氨基酸的含量。方法：样品用含巯基乙酸的盐酸溶液水解提取,经定容、过滤、真空浓缩、复溶、过膜后,用氨基酸分析仪测定,外标法定量。结果：应用含0.05%巯基乙酸的盐酸水解液对奶粉样品进行水解的同时有效防止了蛋氨酸的氧化,相对氧化水解方法节省前处理时间,方法的回收率为94%~97%,相对标准偏差小于3.0%。通过对牛乳基和羊乳基奶粉样品中的16种氨基酸分析,牛乳基奶粉中赖氨酸含量略高于羊乳基奶粉,其他相应氨基酸含量不存在显著性差异。结论：该方法准确快速、重现性好,适用于奶粉中氨基酸含量分析的检测要求。     

淮山药中氨基酸含量的测定

用氨基酸分析仪测定了淮山药中各种氨基酸的组成。淮山药样品经酸水解处理,用氨基酸分析仪进行分析,结果表明:淮山药中含有苏氨酸,缬氨酸,蛋氨酸,苯丙氨酸,异亮氨酸,亮氨酸和赖氨酸等17种氨基酸,总氨基酸质量分数为7.256％,其中人体必需氨基酸的含量占总氨基酸含量的25.32％;说明淮山药在医学和营养学上都有很高的研究价值。     

硒代氨基酸的分析

本文介绍了用日立835-50氨基酸分析仪分析硒代氨酸(硒代胱氨酸、硒代蛋氨酸的方法,由于采用了较高浓度的盐酸水解,中性氨基酸沉淀等技术使两种硒代氨基酸实际样品的同时分析得以实现,国内未见报导。     

淮山药中氨基酸含量的测定

用氨基酸分析仪测定了淮山药中各种氨基酸的组成。淮山药样品经酸水解处理,用氨基酸分析仪进行分析,结果表明：淮山药中含有苏氨酸,缬氨酸,蛋氨酸,苯丙氨酸,异亮氨酸,亮氨酸和赖氨酸等17种氨基酸,总氨基酸质量分数为7．256％,其中人体必需氨基酸的含量占总氨基酸含量的25．32％;说明淮山药在医学和营养学上都有很高的研究价值。     

牛磺酸、羟脯氨酸、γ－氨基丁酸、鸟氨酸和色氨酸的测定

在日立８３５－５０型氨基酸分析仪上,用７２ｍｉｎ程序进行分析,通常一次只能分析１７种氨基酸。采用本文所述方法则一次能分析２２种氨基酸。在不延长分析时间,不增加成本的情况下,一次多分析了五种氨基酸,提高了仪器工作效率。经过精密度、准确度试验和样品分析证明：该方法测定的数据准确、可靠,是一个又快又省的好方法。     

水解方法及氨基酸组成测定

<正> 随着分子生物学、医学、食品工业以及工农业生产的发展,要求氨基酸分析的范围不断扩大而且数量也逐日增多。S.Moore和W.H.Stein首次用离子交换层析法分离蛋白水解液中的氨基酸时分析一个样品需要24小时左右。随着现代     

常用的两种脱色剂对氨基酸定量分析的影响

<正> 在氨基酸分析中常遇到一些样品颜色深,需要进行脱色处理。通常都用活性炭、草酸作脱色剂进行脱色。我们就活性炭、草酸脱色时对氨基酸定量分析的影响进行了比较。具体方法如下:材料:标准氨基酸混合液(日本日立公司);酱油(普通市售二级酱油);活性炭(普通脱色用活性炭,经盐酸处理,烘干备用);草酸(国产二级);日立835～10型带数据处理氨基酸自动分析仪等。     

相思子属三种药材中的氨基酸分析比较

利用全自动氨基酸分析仪和高效液相色谱仪分析比较相思子属三种药材中氨基酸成分及含量。结果9个样品都检测出17种氨基酸和2种色氨酸衍生物。结论:不同样品中的氨基酸含量有所差异,但其中门冬氨酸、谷氨酸、亮氨酸和赖氨酸的含量相对较高。     

Amino acids analysis using a monolithic silica column after derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F)

A fast HPLC method using a monolithic silica column was developed for the measurement of amino acids. The amino acids were pre-column derivatized with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) and separated on a monolithic silica column (MonoClad C18-HS, 250 mm × 3 mm I.D.). The separation of 19 NBD-amino acids was achieved within 18 min, which was only one-fifth of the time taken by the methods using a conventional particle-packed column, with the gradient elution of a mobile phase at the flow rate of 1.4 mL/min. The sensitivity was good with a limit of detection for the individual amino acids ranging from 2.94 to 53.4 fmol. The calibration curves for all the amino acids were found to be linear in the range of 200 fmol to 20 pmol with correlation coefficients of 0.997 or better. The analytical method was successfully applied to determine the amino acids in a mouse plasma sample.     

High-sensitivity amino acid analysis by derivatization with O-phthalaldehyde and 9-fluorenylmethyl chloroformate using fluorescence detection: applications in protein structure determination

An amino acid analysis method using a commercially available analyzer that accurately quantitates protein-derived amino acids in the 10-100 pmol range is described. The method utilizes the robotic capability of the analyzer's autosampler to perform precolumn derivatization of both primary and secondary amino acids with o-phthalaldehyde and 9-fluorenylmethyl chloroformate, respectively. The derivatized amino acids are then separated on a C-18 reverse-phase amino acid column and quantitated in a single run by fluorescence detection. The characterization of beta-lactoglobulin and two tryptic peptides from the bacterial enzyme diaminopimelic acid epimerase is used to demonstrate the sensitivity and utility of this method.     

一种构建高质量随机肽库的有效方法

为构建完全随机化的基因工程肽库 ,克服现有简并方案中终止密码子和序列组成偏歧的不足 ,提出了一种新的简并DNA文库合成方式。通过这种分组式合成方式构建的肽库可以避免终止密码子的出现和氨基酸组成偏歧的发生 ,还可以控制随机化过程中不同氨基酸的参入比例。以一个 13肽库的合成过程为例对分组式合成法进行了实验 ,测序结果和对 19种氨基酸出现频率的统计表明没有终止密码子和半胱氨酸密码子出现 ,各氨基酸的出现频率接近均值 ,表明这种分组 混合 分组 ,辅以简并合成的方法是行之有效的 ,能满足各类高容量基因工程随机肽库要求。     

Development of a protocol for the automated analysis of amino acids in brain tissue samples and microdialysates

An automated precolumn derivatisation method has been developed for the measurement of fourteen amino acids in brain tissue and microdialysate samples. The method involves labelling amino acids with naphthalene-2,3-dicarboxaldehyde (NDA) in the presence of cyanide (CN⁻). The resulting highly stable N-substituted 1-cyanobenz[f]isoindole (CBI) derivatives were separated using a binary gradient elution profile and detected fluorometrically. The order of elution of the derivatised amino acids was confirmed by using liquid chromatography with fluorescence and mass spectrometric detection in tandem. Linear calibration plots were obtained for all amino acids in the range studied (0.2–12.5 μM). The limit of detection for CBI derivatives of amino acids was in the range 5–20 fmol (S/N=2) using a 5 μl injection volume. The method has been used for the measurement of amino acids in microdialysates from rat brain and tissue homogenates from different regions of mouse brain.     

中药鳖甲及两种炮制品的氨基酸营养学

本文报导了鳖甲及其炮制品的氨基酸营养分析情况。结果表明,生品含17种游离氨基酸,传统炮制品含15种游离氨基酸,食用菌炮制品含16种游离氨基酸,且生品总氨基酸的含量明显高于炮制品,经盐酸水解后,炮制前后的样品均测得17种氨基酸,生品的含量略高于炮制品,炮制品前后样品中所测游离氨基酸和水解后氨基酸均含有8种人体必需氨基酸。     

Determination of amino acids in human serum by capillary gas chromatography

A simple and rapid method for the determination of serum amino acids by gas chromatography (GC) has been developed. Following deproteinization of serum with perchloric acid, free amino acids in the supernatant were converted into their N(O,S)-isobutoxycarbonyl methyl ester derivatives and measured by GC with flame ionization detection using a DB-17 capillary column. All the derivatives of the 22 protein amino acids were completely resolved as single peaks within 9 min by GC. The calibration curves were linear in the range 0.2–50 μg of each amino acid, and the correlation coefficients were above 0.998. By using this method, serum amino acids could be directly analysed without prior clean-up procedure such as ion-exchange column chromatography except for deproteinization of the samples, and without any interference from coexisting substances. Overall recoveries of amino acids added to serum samples were 88–108%. Analytical results for serum amino acids from normal subjects are presented.     

Analysis of all protein amino acids as their tert.-butyldimethylsilyl derivatives by gas-liquid chromatography

A gas chromatographic method for the separation and quantitation of the 20 protein amino acids is described using N-methyl-N(tert.-butyldimethylsilyl)trifluoroacetamide, with 1% tert.-butyldimethylchlorosilane as catalyst, to prepare the tert.-butyldimethylsilyl amino acid derivatives. Alkylsilylation of amino acids proceeds at 140 degrees C in 20 min. The derivatives formed in the one-step reaction are used directly for gas-liquid chromatographic analysis, using a flame-ionization detector, without prior isolation or purification. Complete separation and quantitation of all protein amino acids are readily achieved using a 15-m DB-5 capillary column. Strict linearity extends from less than 15 to about 100 ng for all amino acids except Arg, which has a linear range from 50 to 300 ng. The limits of detection, however, range from one to several hundred nanograms. The method was used to analyze the free amino acid pool in carnation petals.     

Analysis of free amino acids during fermentation by Bacillus subtilis using capillary electrophoresis

A high performance capillary electrophoresis (HPCE) method was presented to identify and quantitate free amino acids during fermentation by Bacillus subtilis. Amino acids, pre-column derivatized with phenylisothicyanate, were separated and characterized by HPCE. In order to optimize separation conditions, the assay was developed by varying the β-cyclodextrin concentration and pH of the background electrolyte. A buffer system comprising 30 mM phosphate and 3 mM β-cyclodextrin at pH 7.0, voltage of 20 kV and detection wavelength of 254 nm showed the best results, with 17 out of 20 phenylthioncarbamyl amino acids in a solution adequately separated. For quantification, p-aminobenzoic acid was added as an internal standard. Analysis of free amino acids in Bacillus subtilis culture medium using this method revealed good consistency with the values obtained using conventional ninhydrin-based amino acid analyzer. Four free amino acids (aspartic acid, glutamic acid, proline, and tyrosine) concentration in an extracellular matrix during fermentation by Bacillus subtilis were mainly monitored using this method.     

Quantitative urine amino acid analysis using liquid chromatography tandem mass spectrometry and aTRAQ reagents

Ion-exchange chromatography (IEC) is the most widely used method for amino acid analysis in physiological fluids because it provides excellent separation and reproducibility, with minimal sample preparation. The disadvantage, however, is the long analysis time needed to chromatographically resolve all the amino acids. To overcome this limitation, we evaluated a novel liquid chromatography tandem mass spectrometry (LC-MS/MS) method, which utilizes aTRAQ reagents, for amino acid analysis in urine. aTRAQ reagents tag the primary and secondary amino groups of amino acids. Internal standards for each amino acid are also labeled with a modified aTRAQ tag and are used for quantification. Separation and identification of the amino acids is achieved by liquid chromatography tandem mass spectrometry using retention times and mass transitions, unique to each amino acid, as identifiers. The run time, injection-to-injection, is 25 min, with all amino acids eluting within the first 12 min. This method has a limit of quantification (LOQ) of 1 μmol/L, and is linear up to 1000 μmol/L for most amino acids. The Coefficient of Variation (CV) was less than 20% for all amino acids throughout the linear range. Method comparison demonstrated concordance between IEC and LC-MS/MS and clinical performance was assessed by analysis of samples from patients with known conditions affecting urinary amino acid excretion. Reference intervals established for this method were also concordant with reference intervals obtained with IEC. Overall, aTRAQ reagents used in conjunction with LC-MS/MS should be considered a comparable alternative to IEC. The most attractive features of this methodology are the decreased run time and increased specificity.     

Improved method for the measurement of large neutral amino acids in biological matrices

A high-performance liquid chromatographic method for measuring neutral amino acids in rat sera, brain tissues, and perfusates was developed by using o-phthalaldehyde sulfite as a pre-column derivatization reagent. With the present method, it was possible to separate the neutral amino acids within a single run in 25 min, while the acidic amino acids were eluted near or at the solvent front. The recovery was above 88.8% with a relative standard deviation (RSD) below 4.2%. The within- and between-day assay reproducibility for the determination of rat serum amino acids showed RSDs below 1.35 and 7.61%, respectively. In the present study, the neutral amino acids were assayed with high sensitivity, accuracy and good reproducibility in a relatively short time and on a small sample size.