Fundamental Study on Improvement in Analytical Precision and Speed Associated with Amino Acid Racemization Age-Dating

提高氨基酸外消旋年龄测定相关分析精度和速度的基础研究

• 批准号: 05835009
• 负责人: ABE Iwao
• 金额: $ 1.41万
• 依托单位: College of Engineering, University of Osaka Prefecture
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05835009/
• 关键词: Racemization Dating; Rapid derivatization; Capillary gas chromatography; Aspartic acid; Enantiomer separation; 光学活性固定相; D.L分離; D,L分離

Amino acid racemization dating is known as an unique dating method that can cover the generation field less than 1,000 years and from fifty thousands to hundreds of thousands years which are determination domain outside of radiocarbon and K-Ar dating methods. It is one of advantage that a quantity of fossil sample necessary for 1 determination answers the purpose by approximately 1 g. Howere, weak point of this method is pre-treatment of the fossil sample takes very long time for 1 determination. In this study, the experiment method has been improved for faster dating of the fossils as describing follows. First, fossil has been dissolved in dilute hydrochloric acid of necessary minimum amount. Next, about 5 times amount of ethanol has been added to the hydrochloric acid solution and cooled down to -20ﾟC for 1 hour to precipitate proteins. Collecting the precipitate by means of centrifugation, inorganic salts, free amino acids, low molecular weight peptides, and most of dissolved organi … More c impurities have been removed simultaneously. White precipitate of proteins has been turned into free amino acids by hydrochloric acid hydrolysis. After evaporated off the excess hydrochloric acid, the free amino acids have been derivatized into their N-isopropyloxycarbonyl methyl esters within 1 min. The derivatives have been separated into their enantiomeric pairs by gas chromatography using capillary column coated with modified cyclodextrin as chiral stationary phase. The outcome of this study is as follows : 1) Ion-exchange purification of the hydrolyzed sample could be eliminated, and the time required for derivatization became rapid. Therefore, total analysis time has been shortened by less than 1/2 compared with conventional method. 2) Recoveries of proteins in fossils have increased considerably by employing ethanol precipitation method of proteins in fractionation. According to this method, amount of fossil sample necessary for 1 determination could be reduced to 100 mg result in contribution for preservation of valuable fossil samples. 3) Analytical precision for determination of the D/L ratio of aspartic acid in fossil sample could be hold down to less than 3 % of RSD( relative standard deviation). Less

氨基酸消旋定年法是一种独特的定年方法，它可以覆盖1,000年以下和5万年至10万年的世代范围，是放射性碳和K-Ar定年法之外的测定领域。一个优点是，一次测定所需的化石样品量约为1g。但该方法的缺点是化石前处理时间长。本研究对实验方法进行了改进，以加快化石定年。首先，化石已经溶解在必要的最低量的稀盐酸中。接着，将约5倍量的乙醇加入到盐酸溶液中，并冷却至-20 ℃ 1小时以沉淀蛋白质。通过离心收集沉淀物，无机盐、游离氨基酸、低分子量肽和大部分溶解有机物， ...更多信息 C杂质同时被除去。蛋白质的白色沉淀经盐酸水解转化为游离氨基酸。蒸发掉过量的盐酸后，游离氨基酸已衍生成其N-异丙氧基羰基甲酯在1分钟内。衍生物已被分离成其对映体对气相色谱法使用毛细管柱涂有改性环糊精作为手性固定相。本研究的结果如下：1）水解后的样品可以省去离子交换纯化，衍生化所需的时间变得更快。因此，总分析时间比常规方法缩短不到1/2。2)采用乙醇沉淀法分离蛋白质，大大提高了化石中蛋白质的回收率。根据该方法，1次测定所需的化石样品量可减少至100 mg，有助于保存有价值的化石样品。3)测定化石样品中天冬氨酸D/L比值的分析精密度可控制在相对标准偏差（RSD）小于3%。少

项目成果

I.Abe,N.Fujimoto,N.Nishiyama,K.Terada: "Rapid Analysis of Amino Acid Enantiomers by Chiral Phase Capillary Gas Chromatography" J.Chromatography A. (1996)

I.Abe、N.Fujimoto、N.Nishiyama、K.Terada：“通过手性相毛细管气相色谱快速分析氨基酸对映体”J.Chromatography A. (1996)

Iwao Abe, Masatoshi Kawazuma, Noriteru Fujimoto Taketoshi Nakahara: "N-Alkyloxycarbonyl isobutylamides as readily prepared diamide derivatives of amino acids for separation of enantiomeric isomers by chiral phase capillary gas chromatography" Chem.Lett. N

Iwao Abe、Masatoshi Kawazuma、Noriteru Fujimoto Taketoshi Nakahara：“N-烷氧基羰基异丁基酰胺作为易于制备的氨基酸二酰胺衍生物，用于通过手性相毛细管气相色谱分离对映体异构体”Chem.Lett。

I.Abe,N.Fujimoto,T.Nishiyama,T.Nakahara: "Rapid Derivatization of Amino Acids for Separation of Enantiomeric Isomers by Chiral Phase Capillary Gas Chromatography" Proceeding of the 35th Int.Symp.Chromatography. 713-718 (1995)

I.Abe、N.Fujimoto、T.Nishiyama、T.Nakahara：“通过手性相毛细管气相色谱法快速衍生氨基酸以分离对映体异构体”第 35 届 Int.Symp.Chromatography 论文集。

I.Abe: "2,2,-Trifluoroethyl Chloroformate as a Rapid Derivatizing Reagent ofAmino Acids for Fast Enantiomer Separation by GC" Chemistry Letters. No.2. 113-114 (1995)

I.Abe：“2,2,-三氟乙基氯甲酸酯作为氨基酸的快速衍生试剂，用于通过 GC 进行快速对映体分离”化学快报。

I.Abe: "A New Method for Immobilizationof Chirasil-Val for Gas Chromatography21GC01:Chromatographia" 35. 503-505 (1993)

I.Abe：“气相色谱法固定 Chirasil-Val 的新方法 21GC01：色谱” 35. 503-505 (1993)