Page 229 - Modern Derivatization Methods for Separation Sciences

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Fig. 3.1.
Fluorogenic reactions of primary amines with (A) OPA, (B) NDA, (C) BQCA and (D)
fluorescamine.

2 min intervals, a portion of the resulting mixture is injected into an RP-HPLC system. Extraction with
chloroform or ethyl acetate (100 µl) removes excessive reagent and may be advantageous.

OPA Application: HPLC Assay for l-aminocyclopropanecarboxylic Acid (ACPC) from Plasma and
Brain [12]

Tissue samples (90 µl) from ACPC-treated mice or samples from untreated animals are thawed on the
day of the assay. All samples are spiked with 10 µl of 3.8 mM cycloleucine (internal standard; IS).
Samples are denatured by the addition of 300 µl of 6.2% perchloric acid, followed by 2 min of
centrifugation at 14 000 g. The supernatants are transferred to clean microfuge tubes, placed on ice and
alkalinized by adding 1 ml of 0.4 M KOH. The samples are centrifuged for 2 min at 14000g to remove
precipitated potassium perchlorate. Aliquots of the supernatants (800 µl)

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