Hydroxyproline quantification for the estimation of collagen in tissue using multiple reaction monitoring mass spectrometry

Collagens are highly abundant mammalian proteins that contain a high content of hydroxylated amino acids such as hydroxyproline. We have exploited the high hydroxyproline content of collagen and developed a method for hydroxyproline quantification as a measure of collagen content in muscle samples. The novel method utilizes a highly selective and sensitive method of multiple reaction monitoring (MRM) by mass spectrometry. The analytical method is simple, rapid (5 min), convenient (no derivatization), precise (<17% RSD), accurate (90–108%), sensitive (4.88 nmol/L) and linear (R2 > 0.999) over three orders of magnitude (5–5000 nmol/L).