An　efficient　and　sensitive　method　for　the　separation　and　determination　of　three　essential　amino　acids　and　three　B　vitamins　by　CE-LIF　with　a　simultaneous　derivatization　procedure　was　developed.　The　conditions　for　derivatization　and　separation　of　these　micronutrients　were　investigated.　FITC　was　used　as　the　reagent　for　fluorescence　tagging　of　arginine　(Arg),　valine　(Val),　tryptophan　(Trp),　folic　acid　(FA),　and　niacinamide　(NA).　Riboflavin　(RF)　was　detected　without　derivatization.　Derivatization　of　analytes　dissolved　in　borate　solution　was　performed　by　successive　introduction　of　fluorescence　reagent　and　analytes　followed　by　water　bathing　at　43°C.　The　molar　ratio　of　sample/reagent　(S/R),　derivatization　temperature,　and　incubation　time　significantly　influenced　the　efficiency　of　derivatization.　To　maximize　the　fluorescence　yield,　a　high　S/R　(≥20)　was　required.　The　nonderivatized　RF　and　five　derivatized　analytes　were　separated　in　the　optimized　CE-LIF　system　with　the　application　of　22　kV　voltage　and　25　mM　borate　buffer　at　pH　9.85.　Validation　of　the　method　showed　good　linearity　for　the　corrected　peak　areas　versus　standard　concentrations　for　the　six　analytes.　The　RSDs　(n　=　3)　of　the　migration　time　and　the　peak　area　obtained　for　the　analytes　ranged　from　0.4　to　1.1%　and　from　1.9　to　4.4%,　respectively.　The　developed　method,　with　the　lowest　LOD　of　0.5　nM,　was　successfully　applied　for　the　efficient　derivatization　and　determination　of　B　vitamins　in　four　health　drink　samples.　©　2012　WILEY-VCH　Verlag　GmbH　&　Co.　KGaA,　Weinheim.