{Reference Type}: Journal Article
{Title}: A reliable tool for detecting 7-dehydrocholesterol and cholesterol in human plasma and its use in diagnosis of Smith-Lemli-Opitz syndrome.
{Author}: Luo Y;Liu Z;Zeng Y;Zhang Y;Luan Y;Ma L;Chen L;Zou L;Yang J;Huang Z;Rao Y;Zhang C;
{Journal}: J Sep Sci
{Volume}: 45
{Issue}: 5
{Year}: Mar 2022
{Factor}: 3.614
{DOI}: 10.1002/jssc.202100594
{Abstract}: <B>BACKGROUND: </B>Smith-Lemli-Opitz syndrome is a birth defect caused by the deficiency of 7-dehydrocholesterol reductase in cholesterol biosynthesis pathway, which leads to accumulation of 7-dehydrocholesterol and reduction of cholesterol in body fluids. To effectively diagnose Smith-Lemli-Opitz syndrome and monitor therapy, a reliable method for simultaneous detection of 7-dehydrocholesterol and cholesterol is needed.<BR><B>METHODS: </B>In the presence of antioxidants (2,6-ditert-butyl-4-methylphenol and triphenylphosphine), 50 μL of human plasma were hydrolyzed at 70℃ for 40 min with 1 M potassium hydroxide in 90% ethanol, and then 7-dehydrocholesterol and cholesterol were extracted by 600 μL of n-hexane for three times. After microwave-assisted derivatization with 70 μL of N,O-bis(trimethylsilyl)trifluoroacetamide at 460 W for 3 min, the analytes were measured by gas chromatography-mass spectrometry.<BR><B>RESULTS: </B>The limits of detection were 100 ng/mL for 7-dehydrocholesterol and 300 ng/mL for cholesterol. Good linearity was obtained in the range of 1-600 μg/mL for 7-dehydrocholesterol and 10-600 μg/mL for cholesterol, which completely covered the biochemical levels of Smith-Lemli-Opitz syndrome patients that have been reported.<BR><B>CONCLUSIONS: </B>A time-saving and accurate gas chromatography with mass spectrometry based method was developed for the determination of 7-dehydrocholesterol and cholesterol in human plasma, which also serves as a useful tool for Smith-Lemli-Opitz syndrome diagnosis, treatment, and research.