A routine method for cholesterol and 7-dehydrocholesterol analysis in dried blood spot by GC–FID to diagnose the Smith–Lemli–Opitz syndrome

Gelzo, Monica; Clericuzio, Stefano; Barone, Rosalba; D’Apolito, Oceania; Russo, Antonio Dello; Corso, Gaetano

doi:10.1016/j.jchromb.2012.08.025

Cited by 16 publications

(13 citation statements)

References 21 publications

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“…The analysis in DBS allows to reduce the degree of invasiveness of blood sampling, requires a small amount of sample, and the DBS can be easily sent to laboratories. Several markers are today analyzed in the blood spot to study populations for diagnostic and epidemiological purposes, and not least for drug monitoring (21,(25)(26)(27)(28)(29)(30)(31).…”

Section: Discussionmentioning

confidence: 99%

“…The evaluation of extraction efficiency was performed by a GC‐FID method on three different DBS samples with serum cholesterol levels of 135, 195, and 260 mg/ dl ( n = 15) and analyzed in quintuplicate. The residual cholesterol in DBS was evaluated after the extraction for 30, 60, and 120 min of incubation in methanol.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…The hexane was removed under a gentle stream of N 2 , the dry residue was resuspended in 100 μl of dichloromethane, and 1 μl of solution was injected into GC apparatus. The residual cholesterol in DBS was expressed as percentage of total cholesterol in DBS measured by GC-FID standard method (21). The DBS cholesterol values (n = 317) were correlated with serum cholesterol previously analyzed by automated enzymatic colorimetric method (reference method; Modular Analytics, Roche Diagnostics S.p.A.), and used as training test.…”

Section: Methods Validationmentioning

confidence: 99%

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Development and Validation of an Enzymatic Method for Total Cholesterol Analysis Using Whole Blood Spot

Corso

Papagni

Gelzo

et al. 2015

Clinical Laboratory Analysis

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methods Validationmentioning

confidence: 99%

See 2 more Smart Citations

Development and Validation of an Enzymatic Method for Total Cholesterol Analysis Using Whole Blood Spot

Corso

Papagni

Gelzo

et al. 2015

Clinical Laboratory Analysis

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“…In addition, more and more disorders are being included in newborn screening panels in an effort to ease the diagnostic journey and make disorders amenable to presymptomatic therapies, as they become available. There has been an effort to develop techniques to analyze 7-DHC in dried blood spots [84,85]. However, one of the main problems remains that 7-DHC is not as stable as other diagnostic compounds on dried blood spots, so that pretreated filter paper may be needed [86].…”

Section: Diagnosismentioning

confidence: 99%

Pathogenesis, epidemiology, diagnosis and clinical aspects of Smith–Lemli–Opitz syndrome

Bianconi

Cross

Wassif

et al. 2015

Expert Opinion on Orphan Drugs

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Introduction Smith-Lemli-Opitz Syndrome (SLOS) is a malformation syndrome inherited in an autosomal recessive fashion. It is due to a metabolic defect in the conversion of 7-dehydrocholesterol to cholesterol, which leads to an accumulation of 7-dehydrocholesterol and frequently a deficiency of cholesterol. The syndrome is characterized by typical dysmorphic facial features, multiple malformations, and intellectual disability. Areas covered In this paper we provide an overview of the clinical phenotype and discuss how the manifestations of the syndrome vary depending on the age of the patients. We then explore the underlying biochemical defect and pathophysiological alterations that may contribute to the many disease manifestations. Subsequently we explore the epidemiology and succinctly discuss population genetics as they relate to SLOS. The next section presents the diagnostic possibilities. Thereafter, the treatment and management as is standard of care are presented. Expert opinion Even though the knowledge of the underlying molecular mutations and the biochemical alterations is being rapidly accumulated, there is currently no efficacious therapy addressing neurological dysfunction. We discuss the difficulty of treating this disorder, which manifests as a combination of a malformation syndrome and an inborn error of metabolism. A very important factor in developing new therapies is the need to rigorously establish efficacy in controlled trials.

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A reliable tool for detecting 7‐dehydrocholesterol and cholesterol in human plasma and its use in diagnosis of Smith–Lemli–Opitz syndrome

Luo

Liu

Zeng

et al. 2022

J of Separation Science

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Background 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. Methods 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. Results 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. Conclusion 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.

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A routine method for cholesterol and 7-dehydrocholesterol analysis in dried blood spot by GC–FID to diagnose the Smith–Lemli–Opitz syndrome

Cited by 16 publications

References 21 publications

Development and Validation of an Enzymatic Method for Total Cholesterol Analysis Using Whole Blood Spot

Development and Validation of an Enzymatic Method for Total Cholesterol Analysis Using Whole Blood Spot

Pathogenesis, epidemiology, diagnosis and clinical aspects of Smith–Lemli–Opitz syndrome

A reliable tool for detecting 7‐dehydrocholesterol and cholesterol in human plasma and its use in diagnosis of Smith–Lemli–Opitz syndrome

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