Combined newborn screening for succinylacetone, amino acids, and acylcarnitines in dried blood spots

Coleman Turgeon, Mark J. Magera, Pierre Allard, Silvia Tortorelli, Dimitar Gavrilov, Devin Oglesbee, Kimiyo Raymond, Piero Rinaldo, Dietrich Matern

Research output: Contribution to journalArticle

92 Citations (Scopus)

Abstract

BACKGROUND: Tyrosinemia type I (TYR 1) is a disorder causing early death if left untreated. Newborn screening (NBS) for this condition is problematic because determination of the diagnostic marker, succinylacetone (SUAC), requires a separate first-tier or only partially effective second-tier analysis based on tyrosine concentration. To overcome these problems, we developed a new assay that simultaneously determines acylcarnitines (AC), amino acids (AA), and SUAC in dried blood spots (DBS) by flow injection tandem mass spectrometry (MS/MS). METHODS: We extracted 3/16-inch DBS punches with 300 μL methanol containing AA and AC stable isotope-labeled internal standards. This extract was derivatized with butanol-HCl. In parallel, we extracted SUAC from the residual filter paper with 100 μL of a 15 mmol/L hydrazine solution containing the internal standard 13C 5-SUAC. We combined the derivatized aliquots in acetonitrile for MS/MS analysis of AC and AA with additional SRM experiments for SUAC (m/z 155-137) and 13C 5-SUAC (m/z 160-142). Analysis time was 1.2 min. RESULTS: SUAC was increased in retrospectively analyzed NBS samples of 11 TYR 1 patients (length of storage, 52 months to 1 week;SUACrange, 13-81 μmol/L), with Tyr concentrations ranging from 65 to 293 μmol/L in the original NBS analysis. The mean concentration of SUAC in 13 521 control DBS was 1.25 μmol/L. CONCLUSION: The inclusion of SUAC analysis into routine analysis of AC and AA allows for rapid and cost-effective screening for TYR 1 with no tangible risk of false-negative results.

Original languageEnglish (US)
Pages (from-to)657-664
Number of pages8
JournalClinical Chemistry
Volume54
Issue number4
DOIs
StatePublished - Apr 1 2008

Fingerprint

Screening
Blood
Newborn Infant
Amino Acids
hydrazine
Tyrosinemias
acylcarnitine
succinylacetone
Butanols
Tandem Mass Spectrometry
Isotopes
Mass spectrometry
Methanol
Tyrosine
Assays
Costs and Cost Analysis
Injections
Costs
Experiments

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Combined newborn screening for succinylacetone, amino acids, and acylcarnitines in dried blood spots. / Turgeon, Coleman; Magera, Mark J.; Allard, Pierre; Tortorelli, Silvia; Gavrilov, Dimitar; Oglesbee, Devin; Raymond, Kimiyo; Rinaldo, Piero; Matern, Dietrich.

In: Clinical Chemistry, Vol. 54, No. 4, 01.04.2008, p. 657-664.

Research output: Contribution to journalArticle

Turgeon, C, Magera, MJ, Allard, P, Tortorelli, S, Gavrilov, D, Oglesbee, D, Raymond, K, Rinaldo, P & Matern, D 2008, 'Combined newborn screening for succinylacetone, amino acids, and acylcarnitines in dried blood spots', Clinical Chemistry, vol. 54, no. 4, pp. 657-664. https://doi.org/10.1373/clinchem.2007.101949
Turgeon, Coleman ; Magera, Mark J. ; Allard, Pierre ; Tortorelli, Silvia ; Gavrilov, Dimitar ; Oglesbee, Devin ; Raymond, Kimiyo ; Rinaldo, Piero ; Matern, Dietrich. / Combined newborn screening for succinylacetone, amino acids, and acylcarnitines in dried blood spots. In: Clinical Chemistry. 2008 ; Vol. 54, No. 4. pp. 657-664.
@article{28e733e9fdff494db7a69ad8d70bbd76,
title = "Combined newborn screening for succinylacetone, amino acids, and acylcarnitines in dried blood spots",
abstract = "BACKGROUND: Tyrosinemia type I (TYR 1) is a disorder causing early death if left untreated. Newborn screening (NBS) for this condition is problematic because determination of the diagnostic marker, succinylacetone (SUAC), requires a separate first-tier or only partially effective second-tier analysis based on tyrosine concentration. To overcome these problems, we developed a new assay that simultaneously determines acylcarnitines (AC), amino acids (AA), and SUAC in dried blood spots (DBS) by flow injection tandem mass spectrometry (MS/MS). METHODS: We extracted 3/16-inch DBS punches with 300 μL methanol containing AA and AC stable isotope-labeled internal standards. This extract was derivatized with butanol-HCl. In parallel, we extracted SUAC from the residual filter paper with 100 μL of a 15 mmol/L hydrazine solution containing the internal standard 13C 5-SUAC. We combined the derivatized aliquots in acetonitrile for MS/MS analysis of AC and AA with additional SRM experiments for SUAC (m/z 155-137) and 13C 5-SUAC (m/z 160-142). Analysis time was 1.2 min. RESULTS: SUAC was increased in retrospectively analyzed NBS samples of 11 TYR 1 patients (length of storage, 52 months to 1 week;SUACrange, 13-81 μmol/L), with Tyr concentrations ranging from 65 to 293 μmol/L in the original NBS analysis. The mean concentration of SUAC in 13 521 control DBS was 1.25 μmol/L. CONCLUSION: The inclusion of SUAC analysis into routine analysis of AC and AA allows for rapid and cost-effective screening for TYR 1 with no tangible risk of false-negative results.",
author = "Coleman Turgeon and Magera, {Mark J.} and Pierre Allard and Silvia Tortorelli and Dimitar Gavrilov and Devin Oglesbee and Kimiyo Raymond and Piero Rinaldo and Dietrich Matern",
year = "2008",
month = "4",
day = "1",
doi = "10.1373/clinchem.2007.101949",
language = "English (US)",
volume = "54",
pages = "657--664",
journal = "Clinical Chemistry",
issn = "0009-9147",
publisher = "American Association for Clinical Chemistry Inc.",
number = "4",

}

TY - JOUR

T1 - Combined newborn screening for succinylacetone, amino acids, and acylcarnitines in dried blood spots

AU - Turgeon, Coleman

AU - Magera, Mark J.

AU - Allard, Pierre

AU - Tortorelli, Silvia

AU - Gavrilov, Dimitar

AU - Oglesbee, Devin

AU - Raymond, Kimiyo

AU - Rinaldo, Piero

AU - Matern, Dietrich

PY - 2008/4/1

Y1 - 2008/4/1

N2 - BACKGROUND: Tyrosinemia type I (TYR 1) is a disorder causing early death if left untreated. Newborn screening (NBS) for this condition is problematic because determination of the diagnostic marker, succinylacetone (SUAC), requires a separate first-tier or only partially effective second-tier analysis based on tyrosine concentration. To overcome these problems, we developed a new assay that simultaneously determines acylcarnitines (AC), amino acids (AA), and SUAC in dried blood spots (DBS) by flow injection tandem mass spectrometry (MS/MS). METHODS: We extracted 3/16-inch DBS punches with 300 μL methanol containing AA and AC stable isotope-labeled internal standards. This extract was derivatized with butanol-HCl. In parallel, we extracted SUAC from the residual filter paper with 100 μL of a 15 mmol/L hydrazine solution containing the internal standard 13C 5-SUAC. We combined the derivatized aliquots in acetonitrile for MS/MS analysis of AC and AA with additional SRM experiments for SUAC (m/z 155-137) and 13C 5-SUAC (m/z 160-142). Analysis time was 1.2 min. RESULTS: SUAC was increased in retrospectively analyzed NBS samples of 11 TYR 1 patients (length of storage, 52 months to 1 week;SUACrange, 13-81 μmol/L), with Tyr concentrations ranging from 65 to 293 μmol/L in the original NBS analysis. The mean concentration of SUAC in 13 521 control DBS was 1.25 μmol/L. CONCLUSION: The inclusion of SUAC analysis into routine analysis of AC and AA allows for rapid and cost-effective screening for TYR 1 with no tangible risk of false-negative results.

AB - BACKGROUND: Tyrosinemia type I (TYR 1) is a disorder causing early death if left untreated. Newborn screening (NBS) for this condition is problematic because determination of the diagnostic marker, succinylacetone (SUAC), requires a separate first-tier or only partially effective second-tier analysis based on tyrosine concentration. To overcome these problems, we developed a new assay that simultaneously determines acylcarnitines (AC), amino acids (AA), and SUAC in dried blood spots (DBS) by flow injection tandem mass spectrometry (MS/MS). METHODS: We extracted 3/16-inch DBS punches with 300 μL methanol containing AA and AC stable isotope-labeled internal standards. This extract was derivatized with butanol-HCl. In parallel, we extracted SUAC from the residual filter paper with 100 μL of a 15 mmol/L hydrazine solution containing the internal standard 13C 5-SUAC. We combined the derivatized aliquots in acetonitrile for MS/MS analysis of AC and AA with additional SRM experiments for SUAC (m/z 155-137) and 13C 5-SUAC (m/z 160-142). Analysis time was 1.2 min. RESULTS: SUAC was increased in retrospectively analyzed NBS samples of 11 TYR 1 patients (length of storage, 52 months to 1 week;SUACrange, 13-81 μmol/L), with Tyr concentrations ranging from 65 to 293 μmol/L in the original NBS analysis. The mean concentration of SUAC in 13 521 control DBS was 1.25 μmol/L. CONCLUSION: The inclusion of SUAC analysis into routine analysis of AC and AA allows for rapid and cost-effective screening for TYR 1 with no tangible risk of false-negative results.

UR - http://www.scopus.com/inward/record.url?scp=42449118446&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=42449118446&partnerID=8YFLogxK

U2 - 10.1373/clinchem.2007.101949

DO - 10.1373/clinchem.2007.101949

M3 - Article

VL - 54

SP - 657

EP - 664

JO - Clinical Chemistry

JF - Clinical Chemistry

SN - 0009-9147

IS - 4

ER -