Abstract
In this study, a novel analytical method has been developed for the rapid determination of L-carnitine in human plasma using electrospray ionization tandem mass spectrometry. Free carnitine (FC) was analyzed after extraction with 80% methanol and total carnitine (TC) was analyzed after hydrolysis and extraction. Acyl carnitine (AC) was subtracted FC from TC. Analytical methods used multiple reaction monitoring (MRM) scan modes. A correlation coefficient of linear regression ($r^2$) was 0.9995, recovery was 97%, reproducibility was less than 10%, and limit of detection (LOD) was $0.0016{\mu}mol/L$. This method reduced sample preparation time and showed high resolution and good reproducibility compared to that with liquid chromatographic methods. Normal control showed AC was lower than FC. Clinical management of patients with inborn error of metabolism showed FC was lower than AC. Thus, carnitine fraction level was very important to monitoring patients with metabolic disorder.
본 연구에서는 ESI/MS/MS를 이용하여 혈장에서 카르니틴을 신속하게 정량하는 분석방법을 개발하였다. 분석방법은 free carnitine (FC)는 유도체화 없이 혈액여지를 80% 메탄올로 추출하여 분석하였고, Total carnitine (TC)은 가수분해 한 후 추출하여 multiple reaction monitoring (MRM) scan mode로 분석하였다. Acyl carnitine (AC)는 TC에서 FC를 뺀 값을 사용하였다. 카르니틴의 분석에서 상관계수 ($r^2$)는 0.9995, 회수율은 97%, 재현성은 변동계수가 10% 이하, 검출한계는 $0.0016{\mu}mol/L$ 였다. 이 방법은 기존의 액체 크로마토그래피 보다 전처리가 간다하고 짧은 분석시간과 좋은 감도와 재현성을 보여 주었다. 정상인은 AC가 낮고 FC는 높은 패턴을 보였고, 유전성대사질환 환자는 FC는 낮고 AC가 높은 패턴으로 나타나 카르니틴의 3분획 분석은 유전성대사질환 환자의 확진 및 모니터링에 유용함을 보여주었다.
