I need advice to perform hydroxyproline extraction from bovine and porcine hides, in order to estimate collagen content.
Which is the most efficient and reliable method? I found some literature on internet about acid hydrolysis to extract collagen, but it takes days to complete the extraction. I’ve seen also the alkaline hydrolysis, which takes some hours.
The point is that I don´t need to extract collagen, just the hydroxyproline to determine its concentration (through a modified Woessner method) and then estimate collagen content in hides samples.
Any advice/suggestion/comment will be appreciated.
Best regards.
Dear Mohamed,
Thank you very much for your prompt reply.
Unfortunately, I don't have access to the book chapter.
The paper also describes a 3 days method. I guess that is the only option.
Best regards.
As I understand it, you still have to totally digest the collagen to estimate the hydroxyproline So you are pretty much stuck with acid digestion. Our experience of this was isolating hydroxyproline in bulk for a synthetic application. We used conc HCl in a sealed tube for 24h at 130C.
Exactly, the protocols I've found imply isolating the collagen through acid hydrolysis, and then digestion with proteases. I need a protocol to release the hydroxyproline from the sample directly. If you have an optimized protocol I would greatly appreciate if you can suggest me bibliography.
Pablo, my advisor in Brazil worked a lot with collagen, maybe he or his work can help you. Look for Massami Shimokomaki.
Best regards!
Olá Iris!
I will search for his papers.
Muito Obrigado!
PS. quando vais enviar uns bolos para Argentina!?
Can I do the same protocol to quantify collagen in a broth? Any suggestions. Thank you!
Hi Armarynette, I think you can apply this method to culture media as well. You would need to perform some optimizations with the acid or alkaline hydrolysis before quantifying the hydroxyproline through the Woessner method. Our experiments showed that alkaline is better and faster than acid hydrolysis.
Best regards,
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Enzymology