Determination Of Hydroxyproline In Equine Tendon Using A Modified Spectrophotometric Method
2007
Rassouli, A., Pharmacology and Tocicology Dept, Faculty of Veterinary Medicine, University of Tehran. | Shams, Gh.R., Pharmacology and Tocicology Dept, Faculty of Veterinary Medicine, University of Tehran. | Sharifi, D., Dept of Clinical Sciences, Faculty of Veterinary Medicine. University of Tehran.
Determination of hydroxyproline (OH-Pro) is used as an index for assaying collagen content in connective tissues and in collagen metabolism studies. In this method, tendon samples were hydrolyzed in 6 m HCl, at 105 آ°C for 14-16 hrs and OH-Pro was oxidized by Chloramine T. Then by adding Ehrlich`s reagent and incubating at 60 آ°C, a chromophore was formed. To remove interfering chromophores, OH-Pro product in alkaline media was extracted into toluene and then into acid phase. The absorbance of acid phase was read at 543nm and OH-Pro content was calculated from a calibration curve based on standard solutions run as the same as samples. Calibration curve was linear at 10-160 خ¼g/ml (R2=0.997). The precision of the method defined by CV% of intra- and inter-day variations was less than 7%. Six equine tendon samples were analyzed and their OH-Pro contents were 95-121 mg/g dry weight (109.5 آ± 9.0). This easy, specific and reproducible method can be used practically for determination OH-Pro content in tendon as well as other collageneous tissues like skin.
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