This work suggests an HPLC method for qualitative and quantitative determination of N-epsilon(2-amino-2-carboxyethyl)-L-lysine (LAL). LAL was released from total hydrolysates of various proteins of animal origin and derivatized with dansyl chloride, The performance of two different columns, Spherisorb 3S TG and mu-Bondapack C-18, was compared; better resolution and quantitative response were obtained with the former. The mobile phase was a mixture of 0.01 M phosphate buffer (pH 7) and acetonitrile. Linear response and quantitative repeatability were tested for both detectors used (UV-Vis set at 254 nm; fluorimetric set at lambda(ex(max))= 360 nm and lambda(em(max))= 525 nm). For LAL standard the minimum detectable amount was 0.05 ng, whereas for LAL in actual samples the amount was 0.5 ng (40 mu g/g of analyzed proteins). Good analytical repeatability was obtained, resulting in CV% of 4.7 and 3.8 for UV and fluorimetric detectors, respectively. LAL recovery was determined using both detectors; the values obtained were 94 % (fluorimetric) and 92 % (UV). Greater noise levels were observed with the fluorimetric detector and its higher sensitivity could not, therefore, be fully utilized, The highest amounts of LAL were found in the casein (2816 mu g/g) and cooked albumin (615 mu g/g) samples.
Determination of lysinoalanine by high performance liquid chromatography
MORET, Sabrina;CHERUBIN, Susi;
1994-01-01
Abstract
This work suggests an HPLC method for qualitative and quantitative determination of N-epsilon(2-amino-2-carboxyethyl)-L-lysine (LAL). LAL was released from total hydrolysates of various proteins of animal origin and derivatized with dansyl chloride, The performance of two different columns, Spherisorb 3S TG and mu-Bondapack C-18, was compared; better resolution and quantitative response were obtained with the former. The mobile phase was a mixture of 0.01 M phosphate buffer (pH 7) and acetonitrile. Linear response and quantitative repeatability were tested for both detectors used (UV-Vis set at 254 nm; fluorimetric set at lambda(ex(max))= 360 nm and lambda(em(max))= 525 nm). For LAL standard the minimum detectable amount was 0.05 ng, whereas for LAL in actual samples the amount was 0.5 ng (40 mu g/g of analyzed proteins). Good analytical repeatability was obtained, resulting in CV% of 4.7 and 3.8 for UV and fluorimetric detectors, respectively. LAL recovery was determined using both detectors; the values obtained were 94 % (fluorimetric) and 92 % (UV). Greater noise levels were observed with the fluorimetric detector and its higher sensitivity could not, therefore, be fully utilized, The highest amounts of LAL were found in the casein (2816 mu g/g) and cooked albumin (615 mu g/g) samples.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
