[Objective]To investigate the influence of different factors on the spectrophotometry of hydroxyproline(HYP)in allogeneic tendon.[Methods]The HYP content of allograft tendons was detected by spectrophotometer based on orthogonal test.Four factors,includ-ing enzymolysis time(4 h,12 h,24 h),water bath temperature(37℃,60℃,100℃),acid-enzyme ratio(1∶2,1∶1,1∶1),and water bath oscillation frequency(180 r/min,200 r/min,250 r/min)were set to explore the optimal assay conditions.[Results]The results of single fac-tor analysis showed that under the condition that other variables remained unchanged,the enzymolysis time was 12 h,the water bath temper-ature was 60℃,the acid-enzyme ratio was 2∶1,the pH value was about 7.4,and the shock frequency of 200 r/min could ensure the best enzymolysis effect of HYP.Orthogonal test and range analysis showed that the acid-enzyme ratio had the greatest effect.Other influencing factors are bath temperature,oscillation frequency and time.[Conclusion]In spectrophotometry of HYP content in allograft tendon,pH val-ue,temperature,shock frequency and enzymolysis time should be adjusted to ensure high enzyme activity,so as to achieve the highest de-tection rate of samples,so as to rationally use resources and achieve the best assay efficiency.
allograft tendonhydroxyprolinespectrophotometryorthogonal test
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