Stacking and separation of aspartic acid enantiomers under discontinuous system by capillary electrophoresis with light-emitting diode-induced fluorescence detection.
We describe the stacking and separation of d- and l-aspartic acid (Asp) by capillary electrophoresis (CE) with light-emitting diode-induced fluorescence detection (LEDIF). In the presence of cyanide, d- and l-Asp were derivatized with naphthalene-2,3-dicarboxaldehyde (NDA) to form fluorescent derivatives prior to CE-LEDIF. The separation of NDA-derivatized d- and l-Asp was accomplished using a discontinuous system - buffer vials contained a solution of 0.6% poly(ethylene oxide) (PEO), 150 mM sodium dodecyl sulfate (SDS), and 60mM hydroxypropyl-β-cyclodextrin (Hp-β-CD), while a capillary was filled with a solution of 150 mM SDS and 60mM Hp-β-CD. The role of PEO, Hp-β-CD, and SDS is to act as a concentrating media, as a chiral selector, and as a pseudostationary phase, respectively. This discontinuous system could be employed for the stacking of 600 nL of NDA-derivatized d- and l-Asp without the loss of chiral resolution. The stacking mechanism is mainly based on the difference in viscosity between sample zone and PEO as well as SDS sweeping. The limits of detection at signal-to-noise of 3 for d- and l-Asp were down to 2.4 and 2.5 × 10(-10)M, respectively. Compared to normal sample injection volume (25 nL), this stacking approach provided a 100- and 110-fold improvement in the sensitivity of d- and l-Asp, respectively. This method was further applied for determining d- and l-Asp in cerebrospinal fluid, soymilk, and beer.
Lin
Kai-Cheng
KC
Department of Emergency Medicine, Kaohsiung Veterans General Hospital, Taiwan.
Hsieh
Ming-Mu
MM
Chang
Chia-Wei
CW
Lin
En-Ping
EP
Wu
Tsung-Han
TH
D000818:Animals; D001224:Aspartic Acid / Q000134:cerebrospinal fluid / Q000737:chemistry / Q000302:isolation & purification*; D001515:Beer / Q000032:analysis; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D057230:Limit of Detection; D011092:Polyethylene Glycols / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D041942:Soy Milk / Q000737:chemistry; D013237:Stereoisomerism; D047391:alpha-Cyclodextrins / Q000737:chemistry
D005456:Fluorescent Dyes; D047391:alpha-Cyclodextrins; C479658:hydroxypropyl-alpha-cyclodextrin; D001224:Aspartic Acid; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols