METHOD OF DETERMINING FRACTIONS OF HYDROXYPROLINE IN BIOLOGICAL MATERIAL Russian patent published in 2020 - IPC G01N33/52 

FIELD: medicine.

SUBSTANCE: invention relates to medicine and a method of determining fractions of hydroxyproline in a biological material, comprising collecting biological material, extracting therefrom hydroxyproline by adding ethanol, centrifuging, separating supernatant and precipitate; drying, further dilution in distilled water, separation of diluted supernate into sample 1 to determine fraction of free hydroxyproline and sample 2 to determine fractions of free and peptide-bound hydroxyproline, dilution of precipitate for sample 3 to determine fraction of protein-bound hydroxyproline, adding sodium hydroxide to all samples, conducting alkaline hydrolysis of sample 1 at room temperature and samples 2 and 3 while heating, adding chloramine T as an oxidant to all samples, staining with Ehrlich's reagent, measurement of optical density in all samples, determination of absorbability of free hydroxyproline fraction in sample 1 as per readings of optical density, protein-bound hydroxyproline in sample 3, content of peptide-bound hydroxyproline fraction as difference between content of hydroxyproline in samples 2 and 1, where alkaline hydrolysis is carried out for 25–30 minutes, for its use 4 N sodium hydroxide solution is used, alkaline hydrolysis of samples 2 and 3 is carried out at +120–125 °C; in all samples after alkaline hydrolysis additionally performing neutralization reaction by adding 4 N solution of hydrochloric acid; blood serum is additionally used as biological material, or urine, or lung tissue or spleen homogenate.

EFFECT: invention provides reduced duration of method implementation, high sensitivity of analysis, reduced volume of biological material.

1 cl, 1 dwg, 3 ex