Development of a new measurement method for serum calcium with chlorophosphonazo-III

This version was published on 1 July 2009

Ann Clin Biochem 2009;46:296-301
doi:10.1258/acb.2009.008099
© 2009 Association for Clinical Biochemistry

 

 

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Original Articles


Eisaku Hokazono1,2,
Susumu Osawa1,
Tomota Nakano3,
Yukari Kawamoto3,
Yuji Oguchi3,
Taeko Hotta4,
Yuzo Kayamori4,
Dongchon Kang4,5,
Yuichiro Cho2,
Kiyoko Shiba6 and
Kenji Sato2


1 Department of Health Sciences, Division of Biological Science and Technology, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582;
2 Anatomy and Physiological Science Department of Life Sciences and Bio-informatics, Division of Biomedical Laboratory Sciences, Graduate School of Health Sciences, Tokyo Medical and Dental University, Tokyo;
3 Clinical Department of Isehara Research Laboratory, Technology & Department, Division, Kanto Chemical Co. Inc., Kanagawa;
4 Department of Clinical Chemistry and Laboratory Medicine, Kyushu University Hospital, Fukuoka;
5 Department of Basic Medicine, Faculty of Medical Sciences, Kyushu University, Fukuoka;
6 Department of Clinical Laboratory Medicine, School of Health Science Technology, Bunkyo Gakuin University, Tokyo, Japan


Corresponding author: Eisaku Hokazono, Anatomy and Physiological Science Department of Life Sciences and Bio-informatics, Division of Biomedical Laboratory Sciences, Graduate School of Health Sciences, Tokyo Medical and Dental University, Tokyo, Japan. Email: hokazono{at}shs.kyushu-u.ac.jp


Background: Although serum calcium has been measured using the o-cresolphthaleincomplexone (oCPC) method in the clinical laboratory, this methodstill has some problems regarding linearity and reagent stability.We developed a new measurement procedure using chlorophosphonazo-III(CPZ-III: 2,7-bis (4-chloro-2-phosphonophenylazo) -1,8- dihydroxy-3,6-naphthalenedisulphonic acid, disodium salt) as a chelatorwith an acid medium for serum calcium measurement. The presentmethod showed better linearity and reagent stability comparedwith the oCPC method.

Methods: Characteristics were studied in optimized conditions measuring wavelength by absorption spectra analysis, and interference of protein and metals with Mg2+, Fe2+, Cu2+ and Zn2+. The methodwas applied to an automated analyser (7170; Hitachi High TechnologiesCorp). The measurement performance was evaluated for accuracy,precision, recovery rate, linearity and reagent stability witha comparison study against atomic absorption spectrophotometry(AAS).

Results: The within-run and between-run variations (coefficient of variation [CV]) were 0.92–1.01% and 0.75–1.43%, respectively. The linearity was 0–7.0 mmol/L. The comparison study obtained y = 1.002x (AAS) – 0.10, Sy/x = 0.18 mmol/L, n = 50. Reagent stability was at least 20 d at 4°Cwithout daily calibration.

Conclusion: The new calcium measurement method in serum was demonstratedto have reliable and acceptable performances as a routine testin clinical laboratory.

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