Quantification of cholesterol in human serum by isotope dilution liquid chromatography/mass spectrometry

동위원소희석 액체크로마토그래피/질량분석법에 의한 혈청 내 콜레스테롤의 정량

  • Shin, Hyesun ;
  • Lee, Hwashim ;
  • Lee, Gaeho
  • 신혜선 ;
  • 이화심 ;
  • 이계호
  • Received : 2008.11.10
  • Accepted : 2008.12.12
  • Published : 2008.12.25

Abstract

An ID LC/MS (isotope dilution liquid chromatography/mass spectrometry) was used as a primary method for the quantitative analysis of cholesterol in human serum. The separation of cholesterol was carried out by Thermo ODS hypersil $C^{18}$ column. The mobile phase was 100% methanol, and flow rate was $0.3m{\ell}/min$. Cholesterol and cholesterol-$3,4-13C_2$ were monitored at m/z 369.4 and 371.3, which correspond to $[M-H_2O+H]^+$ respectively. In order to verify the measurement method, NIST SRM 909b was analyzed. The results agreed well with certified values within uncertainty. The four kinds of serum certified reference material were prepared and certified. The repeatabilities of measurement were ranged from 0.1 to 0.8% (RSD), which were relatively good. The reproducibility between independent measurement run was below 0.24% (RSD). The expanded uncertainty was about 1.43% within the 95% confidence interval.

Keywords

cholesterol;ID LC/MS;primary method;traceability

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