Abstract
A liquid chromatography tandem mass spectrometric method is described for the analysis of homocitrulline in human urine, a key metabolite in the differential diagnosis of hyperammonemia, hyperornithinemia, homocitrullinuria (HHH) syndrome. Urine samples were prepared by mere five-fold dilution with a mixture of internal standards (2H2-citrulline and 2H3-creatinine) used for the simultaneous quantification of creatinine. Analytes were separated on a cyano column and eluted isocratically within seven min. Detection was achieved by monitoring transitions of 190 > 84 and 190 > 127 for homocitrulline, 178 > 115 for 2H2-citrulline, 114 > 44 for creatinine and 117 > 47 for 2H3-creatinine. Calibration curves were linear up to 100 micromol/L. Intraday (n = 7) and interday (n = 6) variations were less than 10%. In urine samples from three siblings confirmed to have HHH syndrome, homocitrulline levels were at 13.3 (74), 21.1 (50) and 108.2 (103) mmol/mol creatinine (micromol/L). Control values were 0–9 mmol/mol creatinine (n = 120). The current method solves specificity issues in homocitrulline determination often encountered with some ninhydrin-based systems (coelution with methionine) and some o-phthalaldehyde-based ones (coelution with taurine), and presents an attractive alternative with a relatively high throughput.
Similar content being viewed by others
References
Fernandes J, Saudubray JM, van den Berghe G (eds) (2000) Inborn metabolic diseases. Springer, Berlin Heidelberg New York
Camacho JA, Obie C, Biery B, Goodman BK, Hu C, Almashanu S, Steel G, Gasey R, Lambert M, Mitchell GA, Valle D (1999) Nat Genet 22:151–158
Blau N, Duran M, Blaskovics ME, Gibson KM (eds) (2003) Physician’s guide to the laboratory diagnosis of metabolic diseases. Springer, Berlin Heidelberg New York
Fecarotta S, Parenti G, Vajro P, Zuppaldi A, Casa RD, Carbone MT, Correra A, Torre G, Riva S, Dionisi-Vici C, Santorelli FM, Andria G (2006) J Inherit Metab Dis 29:186–189
Tsujino S, Kanazawa N, Ohashi T, Eto Y, Saito T, Kira J, Yamada T (2000) Ann Neurol 47:625–631
Salvi S, Dionisi-Vici C, Bertini E, Verardo M, Santorelli FM (2001) Hum Mutat 18:460
Miyamoto T, Kanazawa N, Hayakawa C, Tsujino S (2002) Pediatr Neurol 26:65–67
Korman SH, Kanazawa N, Abu-Libdeh B, Gutman A, Tsujino S (2004) J Neurol Sci 218:53–58
Torisu H, Kira R, Kanazawa N, Takemoto M, Sanefuji M, Sakai Y, Tsujino S, Hara T (2006) Brain Dev 28:332–335
Huskova R, Chrastina P, Adam T, Schneiderka P (2004) Clin Chim Acta 350:99–106
Felitsyn NM, Henderson GN, James MO, Stacpoole PW (2004) Clin Chim Acta 350:219–230
Gjessing LR, Lunde HA, Undrum T, Broch H, Alme A, Lie SO (1986) J Inher Metab Dis 9:186–192
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Al-Dirbashi, O.Y., Al-Hassnan, Z.N. & Rashed, M.S. Determination of homocitrulline in urine of patients with HHH syndrome by liquid chromatography tandem mass spectrometry. Anal Bioanal Chem 386, 2013–2017 (2006). https://doi.org/10.1007/s00216-006-0831-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00216-006-0831-5