高效液相色谱法检测鲫鱼不同组织中的胶原蛋白含量

为提高水产品胶原蛋白的利用率，采用柱前衍生-高效液相色谱法对鲫鱼鱼皮、鱼肉、鱼鳞、鱼鳔组织中的羟脯氨酸含量进行检测，再将羟脯氨酸含量换算成相应胶原蛋白的含量。鲫鱼样品经盐酸110 ℃水解18 h，丹磺酰氯衍生，采用高效液相色谱-紫外法进行检测。色谱柱：反相C18柱（250 mm×4.6 mm，5 μm）；流动相：A为甲醇，B为四氢呋喃-甲醇-0.05 mol/L乙酸钠（1∶15∶84，V/V）溶液，采用梯度洗脱；流速1 mL/min；柱温40 ℃；检测波长254 nm。结果表明：羟脯氨酸在20～400 μg/mL范围内呈现良好的关系（R2=0.999 8），最低检测限为0.18 μg/g，不同水平的平均回收率为80.22%～83.33%，相对标准偏差范围为0.31%～1.84%。实际样品分析显示，鱼皮、鱼肉、鱼鳞、鱼鳔组织中胶原蛋白含量分别为106.62、24.75、166.94、115.05 mg/g。本实验建立的分析方法简便快捷、具有较好的灵敏度和可靠性，可用于水产品不同组织中胶原蛋白的定量分析。

Determination of Collagen in Different Tissues of Crucian Carp (Carassius auratus) by High Performance Liquid Chromatography

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.