Abstract:

In order to improve the utilization of collagen in fishery products, a high performance liquid chromatographic
(HPLC) method with pre-column derivatization was established to determine hydroxyproline in the skin, meat, scale, and
blubber of the crucian carp Carassius auratus and the content of collagen was calculated based on hydroxyproline levels
in samples. Samples were hydrolyzed by hydrochloric acid at 110 ℃ for 18 h, followed by pre-column derivatizations
with dansyl chloride. Hydroxyproline was determined by HPLC connected to an ultraviolet detector (HPLC-UV). The
chromatographic conditions were set as follows: column, C18 (250 mm × 4.6 mm, 5 μm); mobile phase, A: methanol, B:
tetrahydrofuran-methanol-0.05 mol/L sodium acetate (1:15:84, V/V); flow rate, 1.0 mL/min; column temperature, 40 ℃; and
detection wavelength, 254 nm. Results showed a good linear relationship existed when hydroxyproline was in the range of
20-400 μg/mL (R2 = 0.999 8). The limit of detection (LOD) was 0.18 μg/g and the average recoveries were 80.22%–83.33%
with relative standard deviations (RSDs) of 0.31%–1.84%. The contents of collagen in skin, meat, scale, blubber of crucian
carp were 106.62, 24.75, 166.94, 115.05 mg/g, respectively. The present method proved to be of high maneuverability,
excellent sensitivity and accuracy, and could be employed to analyze collagen contents in fishery products.

Key words: crucian carp (Carassius auratus), hydroxyproline, collagen, high performance liquid chromatography (HPLC)