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Isocratic Separation of Dansylated Biogenic Amines on a C8‐Bonded Silica Column under the LC Elution of Methanol‐Based Mobile Phase
Authors:Jian‐De Yeh  Shushi Chen
Institution:Department of Applied Chemistry, National Chiayi University, Chiayi, Taiwan 600, R.O.C.
Abstract:Under the elution of methanol‐based mobile phase, the isocratic resolution of 12 biogenic amines, including 1 aromatic, 2 heterocyclic and 9 aliphatic amines, as the dansylated derivatives has been accomplished in less than 25 minutes on a 15 cm C8‐bonded column. The resolution can not be reproduced on other examined alkyl‐bonded phases (e.g., C4 and C18) under the same chromatographic conditions, or in the reversed‐phase mode. The retention, mainly as a result of hydrophobic interaction between analyte and stationary phase, can be adjusted by varying the percentage of methanol in the mobile phase. Also, incorporating acetic acid as additive to the mobile phase to protonate the analyte and silanol groups that are little shielding on the surface of silica gel reduces the dipole‐dipole interaction, and thus the retention scale, which in turn deteriorates the resolution. Furthermore, the elution reversal is plausible for some of analytes as a greater percent of acetic acid is used in the elution. Values of correlation coefficients (R2) range between 0.9995 and 0.9996, indicating good linearity.
Keywords:Biogenic amine  Alkyl‐based phase  HPLC resolution  Chemical derivatization  Isocratic elution  Dansyl derivatives
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