TY - JOUR
T1 - Cation exchange HPLC analysis of desmosines in elastin hydrolysates
AU - Perła-Kaján, Joanna
AU - Gryszczyńska, Agnieszka
AU - Mielcarek, Sebastian
AU - Jakubowski, Hieronim
N1 - Funding Information:
Acknowledgments This work was supported in part by grants from the Ministry of Science and Higher Education, Poland (NN 401 230634) and the American Heart Association (0855919D). We thank Ewa Pruszyńska-Oszmałek (Department of Animal Physiology and Biochemistry, Poznań University of Life Sciences) and Ryszard Słomski (Institute of Human Genetics, Polish Academy of Sciences, Poznań, Poland) for kindly providing mouse and pig organs. We thank Jarosław Zimny for assistance in HPLC analyses in the initial stage of the method development. We also thank anonymous reviewers for their constructive comments.
PY - 2011/11
Y1 - 2011/11
N2 - Desmosine crosslinks are responsible for the elastic properties of connective tissues in lungs and cardiovascular system and are often compromised in disease states. We developed a new, fast, and simple cation exchange HPLC assay for the analysis of desmosine and isodesmosine in animal elastin. The method was validated by determining linearity, accuracy, precision, and desmosines stability and was applied to measure levels of desmosines in porcine and murine organs. The detection and quantification limits were 2 and 4 pmol, respectively. The run-time was 8 min. Our cation exchange column does not separate desmosine and isodesmosine, but their level can be quantified from absorbance at different wavelengths. Using this assay, we found that desmosines levels were significantly lower in elastin isolated from various organs of immunodeficient severe combined immunodeficiency mice compared with wild-type animals. We also found that desmosines levels were lower in lung elastin isolated from hyperhomocysteinemic Pcft -/- mice deficient in intestinal folate transport compared with wild-type Pcft +/+ animals.
AB - Desmosine crosslinks are responsible for the elastic properties of connective tissues in lungs and cardiovascular system and are often compromised in disease states. We developed a new, fast, and simple cation exchange HPLC assay for the analysis of desmosine and isodesmosine in animal elastin. The method was validated by determining linearity, accuracy, precision, and desmosines stability and was applied to measure levels of desmosines in porcine and murine organs. The detection and quantification limits were 2 and 4 pmol, respectively. The run-time was 8 min. Our cation exchange column does not separate desmosine and isodesmosine, but their level can be quantified from absorbance at different wavelengths. Using this assay, we found that desmosines levels were significantly lower in elastin isolated from various organs of immunodeficient severe combined immunodeficiency mice compared with wild-type animals. We also found that desmosines levels were lower in lung elastin isolated from hyperhomocysteinemic Pcft -/- mice deficient in intestinal folate transport compared with wild-type Pcft +/+ animals.
KW - Cation exchange HPLC
KW - Desmosine
KW - Elastin
KW - Homocysteine
KW - Isodesmosine
KW - Pcft mouse
KW - SCID mouse
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U2 - 10.1007/s00216-011-5346-z
DO - 10.1007/s00216-011-5346-z
M3 - Article
C2 - 21887606
AN - SCOPUS:80255129334
SN - 1618-2642
VL - 401
SP - 2473
EP - 2479
JO - Analytical and Bioanalytical Chemistry
JF - Analytical and Bioanalytical Chemistry
IS - 8
ER -