AMINO-ACID-ANALYSIS AND ENZYMATIC SEQUENCE DETERMINATION OF PEPTIDES BY AN IMPROVED ORTHO-PHTHALDIALDEHYDE PRE-COLUMN LABELING PROCEDURE
AMINO-ACID-ANALYSIS AND ENZYMATIC SEQUENCE DETERMINATION OF PEPTIDES BY AN IMPROVED ORTHO-PHTHALDIALDEHYDE PRE-COLUMN LABELING PROCEDURE
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DOI：
10.1080/01483918108059956
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
STEIN, S
中科院分区：
文献类型：
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作者：
JONES, BN;PAABO, S;STEIN, S
伯氨基酸在硫醇存在下与邻苯二甲醛反应形成强荧光衍生物。通过使用反相高效液相色谱法，含有26个这些衍生物的混合物被有效地解析，分析时间< 35分钟。单个氨基酸的定量是可重复的，平均相对偏差为±。1.4%，并且具有约1.0%的检测极限。50毫微微摩尔。通过在衍生化介质中加入十二烷基硫酸钠，提高了赖氨酸和羟基赖氨酸的稳定性和荧光响应。应用色谱系统，包括氨基酸分析的肽后，无论是酸或酶水解。还开发了通过用外肽酶进行时程水解来对pmol量的肽进行序列分析的方法，其采用上述分离程序来鉴定和定量释放的氨基酸。
Primary amino acids react with o-phthaldialdehyde in the presence of mercaptans to form intensely fluorescent derivatives. By the use of reverse-phase high-performance liquid chromatography, a mixture containing 26 of these derivatives was efficiently resolved with an analysis time of < 35 min. The quantitation of the individual amino acids was reproducible with an average relative deviation of .+-.1.4% and had a detection limit of .apprx. 50 femtomol. Improvements in the stability and fluorescence response of lysine and hydroxylysine were obtained by the incorporation of sodium dodecyl sulfate in the derivatizing medium. Applications of the chromatography system involving the amino acid analysis of peptides after either acid or enzymatic hydrolysis are presented. Methods for the sequence analysis of pmol quantities of peptides by time course hydrolysis with exopeptidases were also developed, which employed the above separation procedure for the identification and quantification of the released amino acids.