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• Dried blood spot controls included

• Multiple diagnoses in a single run

• High selectivity and sensitivity by MS/MS detection

• New! Succinylacetone Upgrade Set

The Chromsystems reagent kit for the determination of amino acids and acylcarnitines from dried blood spot samples using tandem mass spectrometry is intended for the early diagnosis of amino acid and fatty acid metabolic disorders during newborn screening.

In Germany, newborn screening is regulated by law. Twelve diseases are defined, eight of them to be determined by tandem mass spectrometry. As therapies for only these specific diseases are known, the diagnosis of other diseases is prohibited. Included in these diseases are defects of amino acid metabolism (phenylketonuria and maple syrup urine disease), defects of fatty acid metabolism (Medium and Very Long Chain Acyl-CoA Dehydrogenase Deficiencies) and organoacidurias (isovaleric acidemia). If not detected, these metabolic errors may impair the newborn severely and irreversibly within the first days of its life.

Specifications (analysis without SUAC):

Specifications (analysis with SUAC):

Cutoff thresholds

LC-MS/MS parameters

LC-MS/MS (tandem MS). No HPLC column is required.

Mass transitions of analytes and internal standards

Sample preparation (general)

• Punch a 3 mm dried blood spot disk out of the filter card into a well of a microtiter plate (or a filter plate).
• Add 200 µl of the reconstituted Internal Standard, seal with a protective sheet and agitate for 20 min at 600 rpm and room temperature.
• Transfer the supernatant into a new microtiter plate. When using a filter plate draw through the supernatant into the well plate by centrifugation (4000 rpm, 2 minutes). Discard the filter plate.
• Evaporate at 60 °C and 600 rpm to dryness.
• Add 60 µl Derivatisation Reagent, seal microtiter plate with a protective sheet and incubate 15 min at 60 °C and 600 rpm.
• Evaporate at 60 °C and 600 rpm to dryness.
• Add 100 µl Reconstitution Buffer and agitate 1 min at 600 rpm.
• Inject 10 µl into the LC-MS/MS system.

Sample preparation (succinylacetone)

• Punch a 3 mm dried blood spot disk out of the filter card into a 96 well plate (or a filter plate).
• Add 200 µl Internal Standard, seal with a protective sheet and agitate for 20 min at 600 rpm and room temperature.
• Transfer the supernatant into a new 96 well plate. When using a filter plate draw through the supernatant into the 96 well plate by centrifugation (4000 rpm, 2 minutes).
• Add 150 µl Internal Standard Succinylacetone to the remaining blood spot and agitate for 30 min at 60 °C.
• Pool the supernatants and evaporate at 60 °C and 600 rpm to dryness.
• Add 60 µl Derivatisation Reagent, seal microtiter plate with a protective sheet and incubate 15 min at 60 °C and 600 rpm.
• Evaporate at 60 °C and 600 rpm to dryness.
• Add 200 µl Reconstitution Buffer and agitate 1 min at 600 rpm.
• Inject 10 µl into the LC-MS/MS system.