Improved analysis of C26:0-lysophosphatidylcholine in dried-blood spots via negative ion mode HPLC-ESI-MS/MS for X-linked adrenoleukodystrophy newborn screening

Haynes, Christopher A.; Jesús, Víctor R. De

doi:10.1016/j.cca.2012.03.026

Cited by 40 publications

(37 citation statements)

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“…The zero enrichment specimen (13104) had a very low measured concentration when analyzed by HPLC–ESI–MS/MS in negative ion mode (0.03 μM) but a higher measured concentration when analyzed by FIA–ESI–MS/MS in positive ion mode (0.7 μM). This result is consistent with previously reports of an isobaric compound in human blood that is observed in positive ion mode but not negative ion mode [12,14,18]. Enrichment with C26:0-LPC was additive with the value measured in specimen 13014, with 100% apparent recovery in the low enrichment specimen (1.7 μM) and 96% apparent recovery in the high enrichment specimen (10.3 μM).…”

Section: Resultssupporting

confidence: 92%

“…The analyte enrichment of human blood utilized commercially available standards prepared as stock solutions (in water or methanol) and their concentrations were verified by comparison of diluted analyte peak areas to commercially available stable-isotope labeled AA, AC, and SUAC. Our results indicated that the recoveries of AA, AC, and C26:0-LPC were similar to the results obtained from two separate methods (AA–AC–SUAC [5] and C26:0-LPC [14]), and that the quantitation of SUAC was slightly less sensitive than NSQAP's in-house AA–AC–SUAC method. Anonymous residual DBS specimens from presumed normal newborns (N = 151) were assayed to determine the mean and 95% confidence interval of C26:0-LPC, and to confirm that AA, AC, and SUAC quantitation was similar to aggregate NBS data from the Region 4 Stork MS/MS Collaborative (R4S) [4,16].…”

Section: Introductionsupporting

confidence: 80%

“…The quantitative results of analysis by the method described here were compared to the results of NSQAP's in-house HPLC analysis [14]. Fig.…”

Section: Resultsmentioning

confidence: 99%

“…C26:0-LPC is a biomarker for X-linked adrenoleukodystrophy (X-ALD) [10] and other peroxisomal biogenesis disorders (PBD). It has previously been analyzed by FIA–ESI–MS/MS [11] and high-performance liquid chromatography–ESI–MS/MS (HPLC–ESI–MS/MS), both of which typically require a second punch from the newborn's DBS sample, a separate extraction protocol, and a separate analysis with respect to AA–AC–SUAC quantitation [12–14]. We used the method described here to quantitate AA, AC, SUAC, and C26:0-LPC in quality control DBS specimens produced at the Centers for Disease Control and Prevention's (CDC) Newborn Screening Quality Assurance Program (NSQAP).…”

Section: Introductionmentioning

confidence: 99%

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Simultaneous quantitation of hexacosanoyl lysophosphatidylcholine, amino acids, acylcarnitines, and succinylacetone during FIA–ESI–MS/MS analysis of dried blood spot extracts for newborn screening

Haynes

Jesús

2016

Clinical Biochemistry

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Objectives The goal of this study was to include the quantitation of hexacosanoyl lysophosphatidylcholine, a biomarker for X-linked adrenoleukodystrophy and other peroxisomal disorders, in the routine extraction and analysis procedure used to quantitate amino acids, acylcarnitines, and succinylacetone during newborn screening. Criteria for the method included use of a single punch from a dried blood spot, one simple extraction of the punch, no high-performance liquid chromatography, and utilizing tandem mass spectrometry to quantitate the analytes. Design and methods Dried blood spot punches were extracted with a methanolic solution of stable-isotope labeled internal standards, formic acid, and hydrazine, followed by flow injection analysis–electrospray ionization–tandem mass spectrometry. Results Quantitation of amino acids, acylcarnitines, and hexacosanoyl lysophosphatidylcholine using this combined method was similar to results obtained using two separate methods. Conclusions A single dried blood spot punch extracted by a rapid (45 min), simple procedure can be analyzed with high throughput (2 min per sample) to quantitate amino acids, acylcarnitines, succinylacetone, and hexacosanoyl lysophosphatidylcholine.

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Section: Resultssupporting

confidence: 92%

Section: Introductionsupporting

confidence: 80%

“…The quantitative results of analysis by the method described here were compared to the results of NSQAP's in-house HPLC analysis [14]. Fig.…”

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Simultaneous quantitation of hexacosanoyl lysophosphatidylcholine, amino acids, acylcarnitines, and succinylacetone during FIA–ESI–MS/MS analysis of dried blood spot extracts for newborn screening

Haynes

Jesús

2016

Clinical Biochemistry

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“…The throughput of a negative ion-mode HPLC–ESI-MS/MS method to detect C26:0-LPC in DBS [7] was improved to achieve an HPLC analysis time of 1 min per sample by increasing the mobile phase flow rate to 0.45 mL/min. The isocratic mobile phase was 50:50 methanol/acetonitrile containing 5 mM ammonium acetate.…”

Section: Methodsmentioning

confidence: 99%

The stability of hexacosanoyl lysophosphatidylcholine in dried-blood spot quality control materials for X-linked adrenoleukodystrophy newborn screening

Haynes

Jesús

2015

Clinical Biochemistry

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Objectives Newborn screening for X-linked adrenoleukodystrophy utilizes tandem mass spectrometry to analyze dried-blood spot specimens. Quality control materials (dried-blood spots enriched with hexacosanoyl lysophosphatidylcholine) were prepared and stored at different temperatures for up to 518 days to evaluate the stability of this biomarker for X-linked adrenoleukodystrophy. Design and methods Dried-blood spot storage included desiccant (45, 171, and 518 days) or omitted desiccant (53 days at >90% relative humidity). Specimens were stored for 171 and 518 days at −20 °C, 4 °C, ambient temperature, and 37 °C. Each weekday for 45 days, a bag of specimens stored at 4 °C was warmed to ambient temperature and one specimen was removed for storage at −80 °C. Specimens were analyzed by high-performance liquid-chromatography electrospray ionization tandem mass spectrometry and data was plotted as concentration (micromoles per liter) vs. time. Linear regression provided slope and y-intercept values for each storage condition. Results Small slope values (0.01 or less) and y-intercept values close to the enrichment indicated less than 11% loss of hexacosanoyl lysophosphatidylcholine under all storage conditions tested. Conclusions Quality control materials for X-linked adrenoleukodystrophy are stable for at least 1 year when stored with desiccant.

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Evaluation of X-Linked Adrenoleukodystrophy Newborn Screening in North Carolina

et al. 2020

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IMPORTANCE X-linked adrenoleukodystrophy (X-ALD) is a peroxisomal genetic disorder in which an accumulation of very long-chain fatty acids leads to inflammatory demyelination in the central nervous system and to adrenal cortex atrophy. In 2016, X-ALD was added to the US Recommended Uniform Screening Panel. OBJECTIVETo evaluate the performance of a single-tier newborn screening assay for X-ALD in North Carolina. DESIGN, SETTING, AND PARTICIPANTSThis diagnostic screening study was of all newborn dried blood spot specimens received in the North Carolina State Laboratory of Public Health between January 2 and June 1, 2018, excluding specimens of insufficient quantity or quality. A total of 52 301 specimens were screened for X-ALD using negative ionization high-performance liquid chromatography tandem mass spectrometry to measure C24:0-and C26:0-lysophosphatidylcholine concentrations. Sanger sequencing of the adenosine triphosphate-binding cassette subfamily D member 1 (ABCD1) gene was performed on screen-positive specimens.EXPOSURES A medical and family history, newborn physical examination, sequencing of ABCD1 on dried blood spot samples, and plasma analysis of very long-chain fatty acids were obtained for all infants with screen-positive results. MAIN OUTCOMES AND MEASURESThe prevalence of X-ALD in North Carolina and the positive predictive value and false-positive rate for the first-tier assay were determined. RESULTSOf 52 301 infants tested (47.8% female, 50.6% male, and 1.7% other or unknown sex), 12 received screen-positive results. Of these 12 infants, 8 were confirmed with a genetic disorder: 3 male infants with X-ALD, 3 X-ALD-heterozygous female infants, 1 female infant with a peroxisome biogenesis disorder, and 1 female infant with Aicardi-Goutières syndrome. Four infants were initially classified as having false-positives results, including 3 female infants who were deemed unaffected and 1 male infant with indeterminate results on confirmatory testing. The positive predictive value for X-ALD or other genetic disorders for the first-tier assay was 67%, with a false-positive rate of 0.0057%. CONCLUSIONS AND RELEVANCEThis newborn screening pilot study reported results on 2 lysophosphatidylcholine analytes, identifying 3 male infants with X-ALD, 3 X-ALD-heterozygous female infants, and 3 infants with other disorders associated with increased very long-chain fatty acids. These results showed successful implementation in a public health program with minimal risk (continued) Key Points Question What is the analytical and clinical validity of a mass spectrometric method evaluating very long-chain fatty acyl-lysophosphatidylcholine species for the detection of X-linked adrenoleukodystrophy among newborns in North Carolina? Findings In this newborn diagnostic screening study of 52 301 dried blood spot specimens, 3 male infants were confirmed to have X-linked adrenoleukodystrophy, 3 female infants were identified as heterozygous for X-linked adrenoleukodystrophy (carriers), 1 female infant had a peroxisome biogenesis...

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Improved analysis of C26:0-lysophosphatidylcholine in dried-blood spots via negative ion mode HPLC-ESI-MS/MS for X-linked adrenoleukodystrophy newborn screening

Cited by 40 publications

References 28 publications

Simultaneous quantitation of hexacosanoyl lysophosphatidylcholine, amino acids, acylcarnitines, and succinylacetone during FIA–ESI–MS/MS analysis of dried blood spot extracts for newborn screening

Simultaneous quantitation of hexacosanoyl lysophosphatidylcholine, amino acids, acylcarnitines, and succinylacetone during FIA–ESI–MS/MS analysis of dried blood spot extracts for newborn screening

The stability of hexacosanoyl lysophosphatidylcholine in dried-blood spot quality control materials for X-linked adrenoleukodystrophy newborn screening

Evaluation of X-Linked Adrenoleukodystrophy Newborn Screening in North Carolina

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