Article
Licensed
Unlicensed Requires Authentication

Comparison of cetylpyridinium chloride and cetyltrimethylammonium bromide extractive procedures for quantification and characterization of human urinary glycosaminoglycans

  • , , , , and
Published/Copyright: May 4, 2010
Clinical Chemistry and Laboratory Medicine
From the journal Clinical Chemistry and Laboratory Medicine Volume 48 Issue 8

Abstract

Background: Glycosaminoglycans (GAGs) are natural complex polysaccharides that are important in several pathological processes. Urinary GAGs have long been investigated for their possible modifications in many pathological conditions. In some cases, they have been found to have diagnostic utility. As a result, the measurement of GAGs in urine is gradually gaining importance. Cetylpyridinium chloride (CPC) and cetyltrimethylammonium bromide (CETAB) are generally used to extract urinary GAGs prior to analysis. In this study, we evaluated the extraction of human urinary GAGs using CPC in comparison with CETAB.

Methods: Extracted urinary GAGs were qualitatively and quantitatively analyzed by agarose-gel electrophoresis in the presence of sequential staining and densitometric scanning. This procedure was able to give more reproducible and reliable results for urinary GAGs, and high performance liquid chromatography (HPLC) was used for the evaluation of chondroitin sulfate (CS) disaccharides.

Results: Differences were observed between CPC and CETAB extract protocols. The absolute amount of CS evaluated by electrophoresis was found to be similar for the two protocols. However, the heparan sulfate (HS) concentration was calculated to be approximately 3.3 times greater for CPC than CETAB. When calculated in relative percentage, 33.6% HS was determined for CPC and 10.0% for CETAB. These results show a quantitative expression for greater recovery of HS by using CPC protocol than CETAB. No significant differences were found between CS quantified by agarose-gel and HPLC. In addition, no differences were observed for the CS disaccharide composition purified by using CPC or CETAB, and quite similar results were observed for 4s/6s disaccharide ratios and charge density values.

Conclusions: Extract procedures for urinary GAGs using CPC or CETAB are able to recover similar amounts of CS quantified by agarose-gel electrophoresis and HPLC. However, CPC yields greater recovery of HS than the CETAB protocol; an increase of approximately 3.3 times as evaluated by electrophoresis. This different capacity of HS extraction between CPC and CETAB should be considered when urinary GAGs of subjects affected by various diseases and related pharmacological treatments are considered, or meta-analysis is performed comparing various studies and trials performed under different experimental conditions.

Clin Chem Lab Med 2010;48:1133–9.

Keywords: chondroitin sulfate; glycosaminoglycans; heparan sulfate; urine analysis
Corresponding author: Prof. Nicola Volpi, Department of Biology, University of Modena and Reggio Emilia, Via Campi 213/D, 41100 Modena, Italy Phone: +39 59 2055543, Fax: +39 59 2055548,
Received: 2009-11-27
Accepted: 2010-2-17
Published Online: 2010-05-04
Published in Print: 2010-08-01

©2010 by Walter de Gruyter Berlin New York

Articles in the same Issue

  1. Editorials
  2. Errors in transfusion medicine are not only misidentifications of the recipient, but also pre-analytical and analytical errors
  3. Biochemical and imaging biomarkers: the search for the Holy Grail
  4. Reviews
  5. Molecular diagnosis of congenital adrenal hyperplasia due to 21-hydroxylase deficiency: an update of new CYP21A2 mutations
  6. Potential role of the lipoxygenase derived lipid mediators in atherosclerosis: leukotrienes, lipoxins and resolvins
  7. Minireview
  8. Errors in transfusion: causes and measures to avoid them
  9. Opinion Paper
  10. Improving the harmonisation of the International Normalized Ratio (INR): time to think outside the box?
  11. Genetics and Molecular Diagnostics
  12. Differential contribution of MTHFR C677T variant to the risk of diabetic nephropathy in Lebanese and Bahraini Arabs
  13. Development and evaluation of an automated hepatitis C virus NS5B sequence-based subtyping assay
  14. A novel mutant-enriched liquidchip technology for the qualitative detection of somatic mutations in KRAS gene from both serum and tissue samples
  15. General Clinical Chemistry and Laboratory Medicine
  16. Urine particle evaluation: a comparison between the UF-1000i and quantitative microscopy
  17. Assessment of the Nova StatSensor whole blood point-of-care creatinine analyzer for the measurement of kidney function in screening for chronic kidney disease
  18. Non-invasive fetal RHD genotyping in the first trimester of pregnancy
  19. Cord blood and maternal serum neopterin concentrations in patients with pre-eclampsia
  20. Comparison of cetylpyridinium chloride and cetyltrimethylammonium bromide extractive procedures for quantification and characterization of human urinary glycosaminoglycans
  21. Increased serum concentrations of visfatin and its production by different joint tissues in patients with osteoarthritis
  22. Human immunodeficiency virus-infection induces major changes in high-density lipoprotein particle size distribution and composition: the effect of antiretroviral treatment and disease severity
  23. A less sensitive detector does not necessarily result in a less sensitive method: fast quantification of 13 antiretroviral analytes in plasma with liquid chromatography coupled with tandem mass spectrometry
  24. Reference Values and Biological Variations
  25. Homocysteine in small-for-gestational age and appropriate-for-gestational age preterm neonates from mothers receiving folic acid supplementation
  26. Cancer Diagnostics
  27. Circulating procalcitonin in aseptic carcinoma patients: a specificity study with 18F-fluorodeoxyglucose positron-emission tomography/computed tomography as benchmark
  28. Increased expression of the oncogenic KLF6-SV1 transcript in human glioblastoma
  29. Thyroglobulin measurement in fine-needle aspirates of lymph nodes in patients with differentiated thyroid cancer: a simple definition of the threshold value, with emphasis on potential pitfalls of the method
  30. Infectious Diseses
  31. Evaluation of the BacT/ALERT® 3D system for the implementation of in-house quality control sterility testing at Canadian Blood Services
  32. Letters to the Editor
  33. Should the external quality assessment sample of antinuclear antibodies be of a typical monospecific pattern?
  34. Identification of reduced serum thrombopoietin concentrations in patients with chronic hepatitis C undergoing an interferon therapy
  35. Plasma glycogen phosphorylase BB is associated with pulmonary artery wedge pressure and left ventricle mass index in patients with hypertrophic cardiomyopathy
  36. Target cell formation leading to pseudoleukocytosis in patients with malignant disorders
  37. Controversies on quotient reporting to standardize laboratory results
  38. Abstracts
  39. Fifth Santorini Conference Biologie Prospective 2010
https://doi.org/10.1515/CCLM.2010.217
Keywords for this article
chondroitin sulfate; glycosaminoglycans; heparan sulfate; urine analysis

Articles in the same Issue

  1. Editorials
  2. Errors in transfusion medicine are not only misidentifications of the recipient, but also pre-analytical and analytical errors
  3. Biochemical and imaging biomarkers: the search for the Holy Grail
  4. Reviews
  5. Molecular diagnosis of congenital adrenal hyperplasia due to 21-hydroxylase deficiency: an update of new CYP21A2 mutations
  6. Potential role of the lipoxygenase derived lipid mediators in atherosclerosis: leukotrienes, lipoxins and resolvins
  7. Minireview
  8. Errors in transfusion: causes and measures to avoid them
  9. Opinion Paper
  10. Improving the harmonisation of the International Normalized Ratio (INR): time to think outside the box?
  11. Genetics and Molecular Diagnostics
  12. Differential contribution of MTHFR C677T variant to the risk of diabetic nephropathy in Lebanese and Bahraini Arabs
  13. Development and evaluation of an automated hepatitis C virus NS5B sequence-based subtyping assay
  14. A novel mutant-enriched liquidchip technology for the qualitative detection of somatic mutations in KRAS gene from both serum and tissue samples
  15. General Clinical Chemistry and Laboratory Medicine
  16. Urine particle evaluation: a comparison between the UF-1000i and quantitative microscopy
  17. Assessment of the Nova StatSensor whole blood point-of-care creatinine analyzer for the measurement of kidney function in screening for chronic kidney disease
  18. Non-invasive fetal RHD genotyping in the first trimester of pregnancy
  19. Cord blood and maternal serum neopterin concentrations in patients with pre-eclampsia
  20. Comparison of cetylpyridinium chloride and cetyltrimethylammonium bromide extractive procedures for quantification and characterization of human urinary glycosaminoglycans
  21. Increased serum concentrations of visfatin and its production by different joint tissues in patients with osteoarthritis
  22. Human immunodeficiency virus-infection induces major changes in high-density lipoprotein particle size distribution and composition: the effect of antiretroviral treatment and disease severity
  23. A less sensitive detector does not necessarily result in a less sensitive method: fast quantification of 13 antiretroviral analytes in plasma with liquid chromatography coupled with tandem mass spectrometry
  24. Reference Values and Biological Variations
  25. Homocysteine in small-for-gestational age and appropriate-for-gestational age preterm neonates from mothers receiving folic acid supplementation
  26. Cancer Diagnostics
  27. Circulating procalcitonin in aseptic carcinoma patients: a specificity study with 18F-fluorodeoxyglucose positron-emission tomography/computed tomography as benchmark
  28. Increased expression of the oncogenic KLF6-SV1 transcript in human glioblastoma
  29. Thyroglobulin measurement in fine-needle aspirates of lymph nodes in patients with differentiated thyroid cancer: a simple definition of the threshold value, with emphasis on potential pitfalls of the method
  30. Infectious Diseses
  31. Evaluation of the BacT/ALERT® 3D system for the implementation of in-house quality control sterility testing at Canadian Blood Services
  32. Letters to the Editor
  33. Should the external quality assessment sample of antinuclear antibodies be of a typical monospecific pattern?
  34. Identification of reduced serum thrombopoietin concentrations in patients with chronic hepatitis C undergoing an interferon therapy
  35. Plasma glycogen phosphorylase BB is associated with pulmonary artery wedge pressure and left ventricle mass index in patients with hypertrophic cardiomyopathy
  36. Target cell formation leading to pseudoleukocytosis in patients with malignant disorders
  37. Controversies on quotient reporting to standardize laboratory results
  38. Abstracts
  39. Fifth Santorini Conference Biologie Prospective 2010
Downloaded on 2.4.2026 from https://www.degruyterbrill.com/document/doi/10.1515/CCLM.2010.217/html
Scroll to top button