MassChrom® Amino Acids and Acylcarnitines from Dried Blood / Non Derivatised - LC-MS/MS

Order No.: 57000/F, for 960 tests
Parameters:
Acylcarnitines & free Carnitine, Amino Acids

Sample prep without derivatisation of the analytes
With 96 well filter plates

Amino Acids and Succinylacetone Acylcarnitines and free Carnitine
Alanine Free Carnitine
Arginine C2-Carnitine
Aspartic acid C3-Carnitine
Citrulline C4-Carnitine
Glutamic acid C5-Carnitine
Glycine C5DC-Carnitine
Leucine C6-Carnitine
Methionine C8-Carnitine
Ornithine C10-Carnitine
Phenylalanine C12-Carnitine
Proline C14-Carnitine
Tyrosine C16-Carnitine
Valine C18-Carnitine
Succinylacetone (included in the Upgrade Set 57111/F)  

Clinical relevance

Newborn screening aims to detect congenital metabolic disorders early on before they can cause harm to the organism. Included in these diseases are defects of amino acid metabolism such as phenylketonuria (PKU) and maple syrup urine disease (MSUD), defects of fatty acid metabolism and organic acidurias (isovaleric acidemia). If these diseases are not diagnosed, the metabolic disorders can already lead to severe irreversible harm to the newborns within their first few days of life.

Mass spectrometry is the method of choice for diagnostic testing: it enables to check for a broad range of metabolic defects in a single run. An HPLC column is not required due to the high selectivity of LC-MS/MS. This makes it possible to achieve a very precise quantification in a very short analysis time of less than 2 minutes.

 

Product advantages

  • Simple SUAC determination with upgrade set
  • Screening for many metabolic disorders in a single run such as PKU (Phenylketonuria), MSUD (Maple Syrup Urine Disease) and tyrosinemia type I
  • Each analyte safeguarded by its own internal standard

 

This Chromsystems assay allows the fast and reliable determination of amino acids, acylcarnitines and succinylacetone from dried blood spot samples as part of the newborn screening for amino acid and fatty acid metabolic disorders using tandem mass spectrometry. Sample preparation is based on effective extraction of the analytes from the blood spot without any further sample preparation steps required. The use of stable isotope labelled internal standards for calibration and measurement ensures reliable and precise quantification of the analytes.

For the analysis of succinylacetone an upgrade set is available (order no. 57111/F).

Here's an overview of all our newborn screening assays.

More Information
Number of Tests 960
Limit of quantification

Amino acids with SUAC: 0.5-12 µmol/l
(Acyl-)carnitines: 0.01-1.9 µmol/l

Linearity

Amino acids with SUAC: 120-7000 µmol/l
(Acyl-)carnitines: 15-400 µmol/l

Recovery Amino acids with SUAC: 41-119 %; (Acyl-)carnitines: 89-103 %
Intraassay

Amino acids with SUAC: CV = 2-10 %
(Acyl-)carnitines: CV = 3-8 %

Interassay

Amino acids with SUAC: CV = 4.3-10 %
(Acyl-)carnitines: CV = 4.3-10.8 %

Analysis Time < 2 min
Cutoff thresholds
Amino acids 30–1090 µmol/l
Free carnitine ca. 55 µmol/l
Acylcarnitines 0.2–73 µmol/l
SUAC 1.2 µmol/l
Sample Preparation
  • Punch out 3.2 mm blood spots from the filter cards into a 96 Well Plate.
  • Add 100 μl Internal Standard and agitate for 20 min.
  • Transfer supernatant into a new 96 Well Plate (or with 57000/F centrifuge for 2 min).
  • Inject up to 10 μl into the LC-MS/MS system.

 

In the sample preparation for the analysis of succinylacetone the following additional steps have to be performed before injection: Add Internal Standard and Extraction Buffer, agitate 30 min, combine extracts (centrifuge), incubate 20 min.

For details please refer to the instruction manual.

Specimen Dried blood
Pre-analytic Treatment

Specimen collection: whole blood from the heel of the newborn taken onto a filter card. Collection between 24th and 72nd hour of life depending on individual national guideline.

Stability: storage at low humidity (< 30 %) and ambient temperature is sufficient if analysis is performed within 24 to 48 hours. For longer storage deep-freeze samples with humidity excluded below -18 °C.

Injection Volume <= 10 µl
Flow rate 20–600 µl/min
MS/MS-Mode MRM
Additional Info

No HPLC column is required.

Information

Mass transitions of analytes and internal standards

Amino acids and SUAC
MRM
Alanine 90 > 44
Alanine-D4 94 > 48
Arginine 175 > 70
Arginine-D7 182 > 77
Aspartic acid 134 > 116
Aspartic acid-D3 137 > 119
Citrulline 176 > 113
Citrulline-D2 178 > 115
Glutamic acid 148 > 130
Glutamic acid-D5 153 > 135
Glycine 76 > 30
Glycine-13C2-15N 79 > 32
Leucine 132 > 86
Leucine-D3 135 > 89
Methionine 150 > 133
Methionine-D3 153 > 136
Ornithine 133 > 70
Ornithine-D6 139 > 76
Phenylalanine 166 > 120
Phenylalanine-D5 171 > 125
Proline 116 > 70
Proline-D7 123 > 77
Tyrosine 182 > 136
Tyrosine-D4 186 > 140
Valine 118 > 72
Valine-D8 126 > 80
SUAC 155 > 137
SUAC-13C5 160 > 142
Acylcarnitines and free carnitine MRM
Carnitine 162 > 85
Carnitine-D9 171 > 85
C2-Carnitine 204 > 85
C2-Carnitine-D3 207 > 85
C3-Carnitine 218 > 85
C3-Carnitine-D3 221 > 85
C4-Carnitine 232 > 85
C4-Carnitine-D3 235 > 85
C5-Carnitine 246 > 85
C5-Carnitine-D9 255 > 85
C5DC-Carnitine 276 > 85
C5DC-Carnitine-D6 282 > 85
C6-Carnitine 260 > 85
C6-Carnitine-D3 263 > 85
C8-Carnitine 288 > 85
C8-Carnitine-D3 291 > 85
C10-Carnitine 316 > 85
C10-Carnitine-D3 319 > 85
C12-Carnitine 344 > 85
C12-Carnitine-D3 347 > 85
C14-Carnitine 428 > 85
C14-Carnitine-D3 375 > 85
C16-Carnitine 400 > 85
C16-Carnitine-D3 403 > 85
C18-Carnitine 428 > 85
C18-Carnitine-D3 431 > 85
Please note The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.
Method of Analysis LC-MS/MS
Parameter Acylcarnitines & free Carnitine, Amino Acids
Autosampler rinsing solution Rinsing Solution (57007)
  1. 57001 newborn screening mobile phase
    Mobile Phase
    Order no.: 57001

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood/Non Derivatised - LC-MS/MS

    Details
  2. 57001 newborn screening mobile phase
    Mobile Phase, 10 x 1000 ml
    Order no.: 57002

     MassChrom® Amino Acids and Acylcarnitines from Dried Blood/Non Derivatised - LC-MS/MS

    Details
  3. 57004 newborn screening internal standard
    Internal Standard
    Order no.: 57004

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood/Non Derivatised - LC-MS/MS

    Details
  4. 57007 newborn screening rinsing solution
    Rinsing Solution
    Order no.: 57007

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood/Non Derivatised - LC-MS/MS

    Details
  5. 57008 newborn screening extraction buffer
    Extraction Buffer
    Order no.: 57008

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood/Non Derivatised - LC-MS/MS

    Details
  6. 96 Well Collection Plate
    96 Well Plates, V-bottomed
    Order no.: 57010

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood/Non Derivatised - LC-MS/MS

    Details
  7. 57057 57010 newborn screening plates
    96 Well Filter Plates
    Order no.: 57057

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood/Non Derivatised - LC-MS/MS

    Details
  8. Protective sheets for 96 Well Plates
    Protective sheets for 96 Well Plates
    Order no.: 55011

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood - LC-MS/MS

    Details
  9. Pierceable adhesive seals for 96 Well Plates
    Pierceable adhesive seals for 96 Well Plates
    Order no.: 55013

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood - LC-MS/MS

    Details
  10. 0191 0192 0193 newborn screening controls
    MassCheck® Amino Acids, Acylcarnitines, Succinylacetone Dried Blood Spot Controls
    Order no.: 0191/0192/0193

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood - LC-MS/MS
    MassChrom® Amino Acids and Acylcarnitines from Dried Blood/Non derivatised - LC-MS/MS

    Details
  1. 0191 0192 0193 newborn screening controls
    MassCheck® Amino Acids, Acylcarnitines, Succinylacetone Dried Blood Spot Controls
    Order no.: 0191/0192/0193

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood - LC-MS/MS
    MassChrom® Amino Acids and Acylcarnitines from Dried Blood/Non derivatised - LC-MS/MS

    Details
  1. Pierceable adhesive seals for 96 Well Plates
    Pierceable adhesive seals for 96 Well Plates
    Order no.: 55013

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood - LC-MS/MS

    Details
  2. Pierceable Heat Seals for 96 Well Plates
    Pierceable Heat Seals for 96 Well Plates
    Order no.: 57014

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood / Non Derivatised - LC-MS/MS

    Details
  3. 57099 LCMS tuning mix amino acids acylcarnitines non derivatised
    Tuning Mix Amino Acids and Acylcarnitines / Non Derivatised
    Order no.: 57099

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood / Non Derivatised - LC-MS/MS

    Details
  4. 57111 newborn screening succinylacetone upgrade set
    Succinylacetone Upgrade Set
    Order no.: 57111/F

    Upgrade for assay no. 57000/F

    Details
  5. Restrictor Capillary
    Restrictor Capillary
    Order no.: 55015

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood - LC-MS/MS

    Details
  6. Adapter collar for centrifugation
    Adapter collar for centrifugation
    Order no.: 55016

    MassChrom® Amino Acids and Acylcarnitines from Dried Blood - LC-MS/MS

    Details
  7. PEEK Prefilter, 2 µm
    PEEK Prefilter, 2 µm
    Order no.: 55033
    Details
  8. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  9. Heat Sealer
    Chromsystems Heat Sealer
    Order no.: 42740

    Easy sealing of 96 well plates
    Adjustable time and temperature
    Ergonomic lever
    Compatible with a wide range of microplates
    Fast startup time
    Small footprint

    Details
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