Hydroxyproline determination in serum and gingival crevicular fluid

Abstract

A high-performance hquid chromatographic assay for total hydroxyproline in serum and gingival crevicular fiuid has been developed. The Clq component of the first complement factor was precipitated out of the samples with a 0.02 M sodium acetate solution. Ahquots of the supernatants were dried, vapor hydrolyzed with 6 M hydrochloric acid at 105°C for 24 hours and derivatized with phenylisothiocyanate. The derivatives were chromatographed using a multilinear solvent gradient with detection at 254 nm. The regression of peak areas on the concentrations of hydroxyproline was linear with a correlation coefficient of 0,99 within the range of assayed quantities (2.5 to 25 ng). Analytical recovery was 90.5 ±2.54% (X + SD) and the determination level was 2.5 ng (19.1 pmol). The precision of the assay as determined by the coefficient of variation for five consecutive runs of six concentrations of hydroxyproline in serum hydrolysate ("within-run" precision) and five series run on different days ("between-run" precision) was 1.8+1.28% and 4.2 + 2.59% (X + SD), respectively. Hydroxyproline concentrations in GCF from single sites during developing experimental gingivitis in the beagle dog showed an irregular pattern of low and high concentrations ranging from 5.2 to 17.4 ng/fA.

Assessment of the alkaline phosphatase level in gingival crevicular fluid, as a biomarker to evaluate the effect of scaling and root planing on chronic periodontitis: An in vivostudy

“…These constituents include physiologically active substances and enzymes such as prostaglandins,[1819] collagenase,[20] hydroxyproline,[21] β-glucuronidase,[22] lactate dehydrogenase,[22] glycosaminoglycans,[23] aspartate aminotransferase,[24] and alkaline phosphatase (ALP). [171925]…”

Elevated levels of collagen cross‐link residues in gingival tissues and crevicular fluid of teeth with periodontal disease

“…Alternative markers of collagen degradation in GCF such as hydroxyproline and the carboxy‐terminal telopeptide of type I collagen (ICTP) have also been evaluated (20–22). The pyridinoline crosslinked ICTP in particular has been suggested to offer a promising option to detect active periodontal destruction (23–27).…”

Hydroxyproline and Total Protein Levels in Gingiva and Gingival Crevicular Fluid in Periodontally Healthy Human Subjects.

“…Gingival crevicular fluid (GCF) carries most of the breakdown products out of tissues, and these subsequently appear in the gingival sulcus [19]. There should be a small amount of GCF even in healthy gingiva, according to the osmotic gradient theory of ALFANO [20,21].…”