食品科学 ›› 2019, Vol. 40 ›› Issue (20): 234-240.doi: 10.7506/spkx1002-6630-20181118-206

• 成分分析 • 上一篇    下一篇

双缩脲法测定罗非鱼源胶原蛋白肽含量的改良及应用评价

张玲,石彩文,肖凯军,李春海,莫军全,张钟,史煜宇   

  1. (1.华南理工大学食品科学与工程学院,广东 广州 510641;2.广东石油化工学院生物与食品工程学院,广东 茂名 525000)
  • 出版日期:2019-10-25 发布日期:2019-10-25
  • 基金资助:
    广东省科技计划项目(2015B020230001);广东普通高校食品科学创新团队项目(2016KCXTD020); 广东石油化工学院2018年大学生创新创业校级培训计划项目(2018pyA038)

ZHANG Ling, SHI Caiwen, XIAO Kaijun, LI Chunhai, MO Junquan, ZHANG Zhong, SHI Yuyu

ZHANG Ling, SHI Caiwen, XIAO Kaijun, LI Chunhai, MO Junquan, ZHANG Zhong, SHI Yuyu   

  1. (1. School of Food Science and Engineering, South China University of Technology, Guangzhou 510641, China;2. College of Biological and Food Engineering, Guangdong University of Petrochemical Technology, Maoming 525000, China)
  • Online:2019-10-25 Published:2019-10-25

摘要: 为探究罗非鱼源胶原蛋白肽含量的快速测定方法,以罗非鱼皮胶原蛋白肽为对照品,改良双缩脲比色法测定肽含量的操作方法,考察显色络合物最大吸收波长、线性拟合度高的肽和三氯乙酸(trichloroacetic acid,TCA)用量范围、pH值;在优化条件下对标准曲线进行重复性、精密度评价以及加样回收率实验;研究本方法对不同分子质量罗非鱼皮及鱼鳞胶原蛋白肽体系检测的适用性。结果显示,当胶原蛋白肽(标称分子质量3 kDa)质量浓度在0.3~1.5 mg/mL范围内,使用13% TCA溶液,调整体系pH 12.5,在波长545 nm处检测,吸光度与肽质量浓度线性拟合好(R2=0.999),标准曲线方程为y=0.144 9x+0.001 5;重复性和精密度实验得到相对标准偏差(relative standard deviation,RSD)分别为1.86%和1.84%,加样回收率分别为113.9%、109.7%,RSD分别为1.39%、1.00%;线性范围宽、重复性好、准确度高。将本法应用于不同种类罗非鱼肽产品体系的检测,加样回收率相差不大,偏差均较小,说明本法简便、快捷,适用于罗非鱼源肽含量的检测。

关键词: 罗非鱼, 胶原蛋白肽, 双缩脲比色法, 肽含量测定, 吸光度

Abstract: To develop a rapid method for the determination of tilapia collagen peptide content, the colorimetric biuret method was improved using tilapia skin collagen peptide as a reference material. We determined the maximum absorption wavelength of the chromogenic complex and the concentration ranges of peptide and trichloroacetic acid (TCA) and pH for high linearity. The repeatability, precision and recovery of the improved method were tested under optimized conditions. The applicability of this method to the detection of collagen peptides from the skin and scales of tilapia with different molecular masses was also studied. The results showed that the absorbance was linearly fitted with peptide concentration ranging from 0.3 to 1.5 mg/mL under the following conditions: 13% TCA, adjustment of the system pH to 12.5, and detection wavelength 545?nm, and the following equation was obtained: y = 0.144 9 x + 0.001 5 (R2 = 0.999). The relative standard deviation (RSD) for repeatability and precision were 1.86% and 1.84%, respectively. The average spiked recoveries with and without the addition of gelatin were 113.9% and 109.7%, with RSD of 1.39% and 1.00%, respectively. The method had wide linear range, good repeatability and high accuracy. Its application to different tilapia collagen peptides showed little differences in spiked recoveries and RSD. The improved biuret method proved to be simple, rapid and suitable for the determination of peptides derived from tilapia.

Key words: tilapia, collagen peptide, colorimetric biuret method, determination of peptide content, absorbance

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