Optimization of Fluorometric Measurement of Free and Total Carnitine in Serum

Muhittin A. SERDAR

Makale detayları

Optimization of Fluorometric Measurement of Free and Total Carnitine in Serum

Yazarlar:

Muhittin A. SERDAR

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Medical Sciences 2001 Vol.31 Issue 4, pp.309-316

Belge Tipi:

Makale

Yayın Tarihi:

3/2001

Belge Dili:

Türkçe

Document ID (Org No.):

2000757266

Sıklık:

İki Aylık

eISSN:

1303-6165

pISSN:

1300-0144

Anahtar kelimeler:

Özet:

 

Carnitine is a major molecule which plays a great role in the transfer of long chain fatty acids into the mitochondrion. Measurement of its total and free forms is essential in the clinical evaluation of carnitine deficiency that can arise from disorders either primarily or secondarily.  In this study, different deproteinization methods (precipitation with perchloric acid, heat denaturation of freeze thawing and filtration) were evaluated for the optimization of fluorometric measurement of total and free carnitine in serum. The most appropriate method for the carnitine measurement was established by additional studies of deacylation at different temperatures, pH optimization of the reaction, tests with reagents of different concentrations and intensity alteration at reaction ends. Recovery of free carnitine and octanoylcarnitine added to serum was 104-92% and 87-91%, respectively. The method was found to be very sensitive for measuring concentrations from 5 to 200 µM. In conclusion, it was observed that there was a significant correlation (r=0.977, slope:1.055, intercept:-2.59) when the optimized method was compared with the UV-kinetic method.

 

Carnitine is a major molecule which plays a great role in the transfer of long chain fatty acids into the mitochondrion. Measurement of its total and free forms is essential in the clinical evaluation of carnitine deficiency that can arise from disorders either primarily or secondarily.  In this study, different deproteinization methods (precipitation with perchloric acid, heat denaturation of freeze thawing and filtration) were evaluated for the optimization of fluorometric measurement of total and free carnitine in serum. The most appropriate method for the carnitine measurement was established by additional studies of deacylation at different temperatures, pH optimization of the reaction, tests with reagents of different concentrations and intensity alteration at reaction ends. Recovery of free carnitine and octanoylcarnitine added to serum was 104-92% and 87-91%, respectively. The method was found to be very sensitive for measuring concentrations from 5 to 200 µM. In conclusion, it was observed that there was a significant correlation (r=0.977, slope:1.055, intercept:-2.59) when the optimized method was compared with the UV-kinetic method.

Makale detayları

Optimization of Fluorometric Measurement of Free and Total Carnitine in Serum

Yazarlar: Muhittin A. SERDAR

Yayıncı: TUBITAK

Dergi: Turkish Journal of Medical Sciences 2001 Vol.31 Issue 4, pp.309-316

Belge Tipi: Makale

Yayın Tarihi: 3/2001

Belge Dili: Türkçe

Document ID (Org No.): 2000757266

Sıklık: İki Aylık

eISSN: 1303-6165

pISSN: 1300-0144

Carnitine

fluorometric measurement

Sayı

Yazarlar:

Muhittin A. SERDAR

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Medical Sciences 2001 Vol.31 Issue 4, pp.309-316

Belge Tipi:

Makale

Yayın Tarihi:

3/2001

Belge Dili:

Türkçe

Document ID (Org No.):

2000757266

Sıklık:

İki Aylık

eISSN:

1303-6165

pISSN:

1300-0144