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

Order No.: 55000/F, for 960 tests
Parameters:
Acylcarnitines & free Carnitine, Amino Acids

Sample prep with derivatisation of the analytes
With 96 well filter plates

Amino Acids
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

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

  • Screening for many metabolic disorders in a single run such as PKU (Phenylketonuria) and MSUD (Maple Syrup Urine Disease)
  • 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 with subsequent derivatisation. The use of stable isotope labelled internal standards for calibration and measurement ensures reliable and precise quantification of the analytes.

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

More Information
Number of Tests 960
Limit of quantification

Amino acids: 0.5-5 µmol/l
(Acyl-)carnitines: 0.05-0.5 µmol/l

Linearity

Amino acids: 250-2500 µmol/l
(Acyl-)carnitines: 11-240 µmol/l

Recovery Amino acids: 51-94 %; (Acyl-)carnitines: 62-89 %
Intraassay

Amino acids: CV = 4-10 %
(Acyl-)carnitines: CV = 4-20 %

Interassay

Amino acids: CV = 5-9 %
(Acyl-)carnitines: CV = 8-28 %

Analysis Time < 2 min
Cutoff thresholds
Amino acids 22-743 µmol/l
Free carnitine 50-60 µmol/l
Acylcarnitines 0.1-48 µmol/l
Sample Preparation
  • Punch out 3.2 mm blood spots from the filter cards into a 96 Well Plate.
  • Add 200 μl Internal Standard and agitate for 20 min.
  • Transfer supernatant into a new 96 Well Plate (or with 55000/F centrifuge for 2 min).
  • Evaporate supernatant.
  • Add 60 μl Derivatisation Reagent and incubate for 15 min at 60 °C.
  • Evaporate to dryness.
  • Add 100 μl Reconstitution Buffer and agitate for 1 min.
  • Inject 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 derivatisation: Add Internal Standard, agitate 30 min, combine extracts (pipette).

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 Amino acids: MRM / Neutral Loss Scan of 102 (m/z); Acylcarnitines: Parent Ion Scan of 85 (m/z)
Additional Info

No HPLC column is required.

Information

Mass transitions of analytes and internal standards

Amino acids Neutral Loss Scan 102
Alanine 146 > 44
Alanine-D4 150 > 48
Aspartic acid 246 > 144
Aspartic acid-D3 249 > 147
Glutamic acid 260 > 158
Glutamic acid-D5 265 > 163
Leucine 188 > 86
Leucine-D3 191 > 89
Methionine 206 > 104
Methionine-D3 209 > 107
Phenylalanine 222 > 120
Phenylalanine-D5 227 > 125
Tyrosine 238 > 136
Tyrosine-D4 242 > 140
Valine 174 > 72
Valine-D8 182 > 80
Amino acids MRM
Arginine 231 > 70
Arginine-D7 238 > 77
Citrulline 232 > 113
Citrulline-D2 234 > 115
Glycine 132 > 76
Glycine-13C2-15N 135 > 79
Ornithine 189 > 70
Ornithine-D6 195 > 76
Proline 172 > 116
Proline-D7 179 > 123
Acylcarnitines and free carnitine Parent Ion Scan 85
Carnitine 218 > 85
Carnitine-D9 227 > 85
C2-Carnitine 260 > 85
C2-Carnitine-D3 263 > 85
C3-Carnitine 274 > 85
C3-Carnitine-D3 277 > 85
C4-Carnitine 288 > 85
C4-Carnitine-D3 291 > 85
C5-Carnitine 302 > 85
C5-Carnitine-D9 311 > 85
C5DC-Carnitine 388 > 85
C5DC-Carnitine-D6 394 > 85
C6-Carnitine 316 > 85
C6-Carnitine-D3 319 > 85
C8-Carnitine 344 > 85
C8-Carnitine-D3 347 > 85
C10-Carnitine 372 > 85
C10-Carnitine-D3 375 > 85
C12-Carnitine 400 > 85
C12-Carnitine-D3 403 > 85
C14-Carnitine 428 > 85
C14-Carnitine-D3 431 > 85
C16-Carnitine 456 > 85
C16-Carnitine-D3 459 > 85
C18-Carnitine 484 > 85
C18-Carnitine-D3 487 > 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 (55007)
  1. 55001 newborn screening mobile phase
    Mobile Phase
    Order no.: 55001

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

    Details
  2. 55002 newborn screening mobile phases
    Mobile Phase, 10 x 1000 ml
    Order no.: 55002

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

    Details
  3. 55004 newborn screening internal standard
    Internal Standard
    Order no.: 55004

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

    Details
  4. 55005 newborn screening derivatisation reagent
    Derivatisation Reagent
    Order no.: 55005

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

    Details
  5. 55006 newborn screening reconstitution buffer
    Reconstitution Buffer
    Order no.: 55006

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

    Details
  6. 55007 LCMS amino acid acylcarnitines rinsing solution
    Rinsing Solution
    Order no.: 55007

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

    Details
  7. 55008 newborn screening extraction buffer
    Extraction Buffer
    Order no.: 55008

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

    Details
  8. newborn screening
    96 Well Plates
    Order no.: 55010/F

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

    Details
  9. 55057 55010 newborn screening
    96 Well Filter Plates
    Order no.: 55057

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

    Details
  10. 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
  11. 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. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  2. 55099 LCMS tuning mix amino acids acylcarnitines
    Tuning Mix Amino Acids and Acylcarnitines
    Order no.: 55099

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

    Details
  3. 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
  4. Restrictor Capillary
    Restrictor Capillary
    Order no.: 55015

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

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

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

    Details
  6. PEEK Prefilter, 2 µm
    PEEK Prefilter, 2 µm
    Order no.: 55033
    Details
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