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使用高效液相色谱法进行氨基酸分析的柱前衍生化已成为一种使用选择性、高灵敏度检测器和反相柱同时分离多种成分的方法。使用 dabsyl chloride (4-(Dimethylamino)azobenzene-4’-sulfonyl chloride:DABS-Cl) 作为柱前衍生试剂进行氨基酸分析的许多结果已经被报道,因为dabsyl chloride 衍生的氨基酸比其他衍生剂衍生的氨基酸更稳定。此外,可以使用具有高灵敏度的简单的UV/VIS检测器。在这里,氨基酸的标准混合物是通过使用dabsyl chloride 的系统测量的Application Note 2/3Application NoteAnalysis of Dabsyl Amino Acids using HPLC Analysis of Dabsyl Amino Acids using HPLC The pre -column derivatiz a tion using HPLC for amino acids analysis has prevailed as a simultaneous separation method of multiple components using a selective , high-sensi t ivity detector and a reversed phase column. Many results of amino acids analysis u s ing dabsyl chloride (4-(Dimethylamino)azobenzene -4'-sulfonyl chlor i de:DABS-CI) as a reagent for pre-column derivatization have been reported because th e amino ac i ds derivatized by dabsyl chloride are more stable than those by other derivatization agents. Also a s i mply a UV/IS detector can be used with high sensitivity. Here the standard mixture of amino acids was measured by the system using dabsyl chloride Keyword: HPLC , Amino acids, DABS-CI , DAB Label ,5.0 pm, Dabsy l pak I, UV/IS detector Jasco PU-2080, DG-2080-53,LG-2080-02,AS-2055,CO-2060,UV-2070 Experimental Equipment: Conditions: Experimental Equipment: Pump: PU-2080 Degasser: DG-2080-53 Mixer: LG-2080-02 Column oven: AS-2055 Autosampler: CO-2060 Detector: UV-2070 Column: Dabsylpak ⅡI (4.6 mmlD x 150 mmL, 5um) Pre-column: Dabsylpak II-P (4.6 mmlDx35 mmL, 5 pm) Eluent A: 20 mM Sodium acetate (pH6.0) Eluent B: Acetonitrile Gradient condition: (A/B), 0 min (78/22, 3 min (78/22),25 min (70/30), 40 min (40/60),40.1 min (20/80), 45 min (20/80) , 45.1 min (78/22), 1cycle: 55min Flow rate: 1.0 mL/min Column temp.: 45℃ Wavelength: 465 nm Injection volume: 20 pL Standard sample: Amino acids mixture 40 pmol each, Type H Fi g u r e 1 sh o w s t h e c hr o ma t o gram o f s t a n da rd mi xt u r e o f a m in o a c i ds. 17 k i nds o f a m in o a c i ds w e r e se pa rat e d i n 45 m inu t es . Figure 1. C h romatogra m of standard m ixture o f amin o acids. Aspar t ic ac i d , Glutami c a c id, Serine,Threo n ine, Arginine, Glycine, Alanin e , P rolin e , Valine, Met h ionin e , I soleuc i ne, L euc i ne, P h e n ylalan i ne,Cys t in e , Lysi n e , H ist i d i ne, T yrosi n e A d v a n ta g e s o f Da b s y l a tio n - B y usi n g t he k i t (D A B L ab e l ), t he p roc ed u r e o f i n tr i ca t e p r e p a r a ti o n o f d er i v a t i z ation ca n be av o id ed .- Amin o a c i d c o m p o s i t ion o f 0.5 p g o f p r o t ei n s o r p e pti d e s c an a n a l y ze d w i th go o d acc u r a cy a n d r e p r o duci bi l i t y. - 17 k in ds o f t he de ri va t i z ed a min o a cids ca n b e s epa r at e d w i th i n 45 min u t e s . D A BS -C I r eact s w i t h a-a m i n o g r ou p s,8a mi n o g r o up s, p h en ol ic h y dr ox yl g r o up s a n d i m i da zole g r o u p s .F i gure 2 sh o ws a r ea c ti o n f o r mu l a wit h a-a m i n o gr oup .
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佳士科商贸有限公司为您提供《Dabsyl氨基酸的高效液相色谱分析》,该方案主要用于其他中氨基酸、检测、分析检测,参考标准《暂无》,《Dabsyl氨基酸的高效液相色谱分析》用到的仪器有JASCO高效色谱仪LC-4000。
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