Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Search research articles

Related Experiment Videos

Total urinary hydroxyproline determined with rapid and simple high-performance liquid chromatography.

R Paroni¹, E De Vecchi, I Fermo

¹Istituto Scientifico H. San Raffaele, Milano, Italy.

Clinical Chemistry

|March 1, 1992

Summary

This study optimized a rapid HPLC method for analyzing total urinary hydroxyproline. The new assay is specific, accurate, and correlates well with existing methods, offering a reliable tool for clinical diagnostics.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Comparison of multiple definitions for ventilator-associated pneumonia in patients requiring mechanical ventilation for non-pulmonary conditions: preliminary data from PULMIVAP, an Italian multi-centre cohort study.

The Journal of hospital infection·2023

Same author

Sphingolipid composition of circulating extracellular vesicles after myocardial ischemia.

Scientific reports·2020

Same author

Can leucocyte esterase replace frozen sections in the intraoperative diagnosis of prosthetic hip infection?

The bone & joint journal·2019

Same author

Modulation of opportunistic species Corynebacterium diphtheriae, Haemophilus parainfluenzae, Moraxella catarrhalis, Prevotella denticola, Prevotella melaninogenica, Rothia dentocariosa, Staphylococcus aureus and Streptococcus pseudopneumoniae by intranasal administration of Streptococcus salivarius 24SMBc and Streptococcus oralis 89a combination in healthy subjects.

European review for medical and pharmacological sciences·2019

Same author

Hyaluronic Acid and Its Composites as a Local Antimicrobial/Antiadhesive Barrier.

Journal of bone and joint infection·2017

Same author

Leucocyte esterase, glucose and C-reactive protein in the diagnosis of prosthetic joint infections: a prospective study.

Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases·2016

Area of Science:

Analytical Chemistry
Biochemistry
Clinical Chemistry

Background:

Hydroxyproline is a key indicator of collagen turnover.
Accurate measurement of urinary hydroxyproline is crucial for diagnosing bone and connective tissue disorders.
Existing analytical methods can be time-consuming or lack specificity.

Purpose of the Study:

To develop and optimize a rapid and specific precolumn derivatization method for total urinary hydroxyproline analysis using High-Performance Liquid Chromatography (HPLC).
To validate the performance characteristics of the optimized HPLC assay, including linearity, detection limit, recovery, and precision.
To compare the results obtained from the new HPLC method with established techniques like ion-exchange chromatography and a colorimetric procedure.

Main Methods:

Related Experiment Videos

Urine samples underwent overnight hydrolysis.
Samples were derivatized using N,N-diethyl-2,4-dinitro-5-fluoroaniline (FDNDEA) after reconstitution with cysteic acid.
Separation was achieved on an Ultrasphere ODS column with a specific mobile phase, and detection was at 360 nm.

Main Results:

The optimized HPLC method provided a rapid analysis with a run time of 18 minutes and a hydroxyproline retention time of 7.3 minutes.
The assay demonstrated linearity from 5 to 100 mg/L with a low detection limit (0.8 ng injected, equivalent to 2 mg/L in urine).
High analytical recovery (94.2-104%) and good precision (CVs < 4.33%) were observed. The HPLC method showed excellent correlation with ion-exchange chromatography (r=0.985) and good correlation with a colorimetric method (r=0.937).

Conclusions:

The developed precolumn derivatization HPLC method is a rapid, specific, and reliable technique for quantifying total urinary hydroxyproline.
This method offers advantages in terms of speed and specificity compared to traditional methods.
The strong correlation with established techniques validates its utility for clinical and research applications in assessing collagen metabolism.