Page 159 - Modern Derivatization Methods for Separation Sciences

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[42].

PITC Application: Derivatization of Amino Acids [43]

Standards or sample preparations which had been evaporated to dryness were treated with 10 µ1 of a
mixture of methanol-1 M sodium acetate -triethylamine (TEA) (2:2:1, v/v) and the drying process was
repeated under vacuum until a pressure of 70 mTorr was reached. This redrying step ensured that any
residual acid was neutralized and that the alkaline pH which favored derivatization was attained. The
derivatization reagent consisted of MeOH-TEAwater-PITC (7:1:1:1, v/v) and was freshly prepared each
time. The dried sample was dissolved in 20 µ1 of derivatizing reagent and left to react for 20 min at
ambient temperature. Excess reagent, TEA and

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