英文名称： 55000 MassChrom® Amino Acids and Acylcarnitines, Reagent kit for LC-MS/MS analysis
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):
| Linearity: | Amino acids: 2000-2400 µmol/l (Acyl-)carnitines: 25-200 µmol/l |
| Limit of quantification: | Amino acids: 2-16 µmol/l (Acyl-)carnitines: 0.1-1.6 µmol/l |
| Recovery: | Amino acids: 67-90 % (Acyl-)carnitines: 81-97 % |
| Intra-assay: | Amino acids: 3-9 % (Acyl-)carnitines: 4-16 % |
| Inter-assay: | Amino acids: 6-11 % (Acyl-)carnitines: 7-20 % |
| Analysis time: | < 2 min |
Specifications (analysis with SUAC):
| Linearity: | Amino acids with SUAC: 250-2500 µmol/l (Acyl-)carnitines: 11-240 µmol/l |
| Limit of quantification: | Amino acids with SUAC: 0.5-5 µmol/l (Acyl-)carnitines: 0.05-0.5 µmol/l |
| Recovery: | Amino acids with SUAC: 51-94 % (Acyl-)carnitines: 62-89 % |
| Intraassay: | Amino acids with SUAC: 4-10 % (Acyl-)carnitines: 4-20 % |
| Interassay: | Amino acids with SUAC: 5-9 % (Acyl-)carnitines: 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 |
| SUAC: | 1.16 µmol/l |
LC-MS/MS parameters
LC-MS/MS (tandem MS). No HPLC column is required.
| Injection volume: | 10 µl |
| Flow gradient: | 20-600 µl/min |
| Autosampler rinsing solution: | Rinsing Solution (55007) |
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 |
| 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 |
| SUAC | 211 > 109 |
| SUAC-13C5 | 216 > 114 |
| (Acyl-)carnitines | 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 |
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.
| Order no. | Product |
|---|---|
| 55000 | MassChrom® Reagent kit for the determination of Amino Acids and Acylcarnitines from dried blood spot samples (LC-MS/MS) with regular 96 well plates. For 960 analyses. |
| 55000/F | MassChrom® Reagent kit for the determination of Amino Acids and Acylcarnitines from dried blood spot samples (LC-MS/MS) with 96 well filter plates. For 960 analyses. |
| Separately available components | |
| 55001 | Mobile Phase, 1000 ml |
| 55002 | Mobile Phase, 10 x 1000 ml |
| 55004 | Internal Standard (lyoph.), 4 x 50 ml |
| 55044 | Internal Standard for Succinylacetone, 3 x 50 ml |
| 55004/T | Internal Standard (lyoph.), 50 ml |
| 55044/T | Internal Standard for Succinylacetone, 50 ml |
| 55005 | Derivatisation Reagent, 30 ml |
| 55006 | Reconstitution Buffer, 100 ml |
| 55007 | Rinsing Solution, 1000 ml |
| 55008 | Extraction Buffer, 200 ml |
| 55010 | 96-Well-Plates, 10 pcs. |
| 55011 | Protective sheets for 96 well plates, 10 pcs. |
| 55057 | 96-Well-Filter-Plates, 5 pcs. |
| Succinylacetone Upgrade Set | |
| 55111 | Succinylacetone Upgrade Set for 960 analyses, consisting of: |
| Internal Standard for Succinylacetone, 3 x 50 ml | |
| Reconstitution Buffer, 100 ml | |
| Startup accessories | |
| 55013 | Pierceable adhesive seal for 96 well plates, 10 pcs. |
| 55015 | Restrictor Capillary, 1 pc. |
| 55016 | Adapter collar for centrifugation, 2 pcs. |
| 55098 | Tuning Mix for Succinylacetone, 1 ml |
| 55099 | Tuning Mix incl. all analytes, 2 ml |
| 55033 | PEEK prefilter, 5 µm, 5 pcs. |
| 15010 | PEEK prefilter housing, 1 pc. |
| Chromsystems quality control material (filter spot): | |
| 0191 | MassCheck® Amino Acids, Acylcarnitines Dried Blood Spot Control, Bi-Level (I+II), 2 x 3 Spots |
| 0192 | MassCheck® Amino Acids, Acylcarnitines Dried Blood Spot Control, Level I, 1 x 3 spots |
| 0193 | MassCheck® Amino Acids, Acylcarnitines Dried Blood Spot Control, Level II, 1 x 3 spots |