Simultaneous analysis of lysine, Nɛ-carboxymethyllysine and lysinoalanine from proteins

Protein quality was assayed by simultaneous measurement of lysine (Lys), carboxymethyllysine (CML) and lysinoalanine (LAL). GC-FID analysis of N-tert-butyl dimethylsilyl (tBDMSi) derivatives of these amino acids was undertaken. tBDMSi derivates were separated on a CP-SIL 5CB commercially fused silica capillary column (25 m × 0.25 mm i.d., 0.25 μm film thickness) employing a thermal gradient programmed from 200 to 300 °C. The identity of tBDMSi derivatives of Lys, CML and LAL was established by GC–MS while FID detection was employed for quantification. Analytical parameters such as linearity (lysine 350–4200 μM, LAL 3–81 μM, CML 16–172 μM), precision (1–13% variation coefficients), accuracy …