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22 Acylcarnitines and 13 Amino acids (Endogenous) in Dried Blood Spots with 16 Deuterated Internal Standards on Raptor HILIC-Si EXP Guard Cartridge Column by LC-MS/MS

PeakstR (min)Precursor IonProduct Ion
1.Phenylalanine0.20166.0120.1
2.Leucine0.21132.186.0
3.Leucine-d30.21135.289.1
4.Isoleucine0.21132.186.1
5.Tyrosine0.22182.191.0
6.Methionine0.24150.156.1
7.Methionine-d30.24153.2107.1
8.Valine0.27118.172.0
9.C20-Eicosanoyl-L-carnitine0.27456.485.1
10.C18-Stearoyl-L-carnitine0.28428.385.1
11.C18-Stearoyl-L-carnitine-d30.29431.485.1
12.C18:1 Oleoyl-L-carnitine0.29426.485.1
13.C18:2 Linoleoyl-L-carnitine0.30424.385.1
14.C16-Palmitoyl-L-carnitine0.30400.385.1
15.C16-Palmitoyl-L-carnitine-d30.31403.385.1
16.C16:1 Palmitolelyl-L-carnitine0.31398.385.1
17.C14-Myristoyl-L-carnitine0.32372.385.1
18.C14-Myristoyl-L-carnitine-d30.33375.385.1
19.C14:1 Tetradecenoyl-L-carnitine0.33370.385.1
20.C14:2-Tetradecadienoyl-L-carnitine0.33368.385.1
21.Proline0.33116.070.1
22.C12-Lauroyl-L-carnitine0.35344.385.1
23.C12-Lauroyl-L-carnitine-d30.35347.385.1
24.Alanine0.3690.144.1
25.Alanine-d40.3594.148.1
26.C10-Decanoyl-L-carnitine0.38316.385.1
27.C10-Decanoyl-L-carnitine-d30.39319.285.1
28.C8-Octanoyl-L-carnitine0.43288.385.1
29.C8-Octanoyl-L-carnitine-d30.43291.285.1
30.C7-Heptanoyl-L-carnitine0.45274.285.1
31.C6-Hexanoyl-L-carnitine-d30.46263.285.1
32.C6-Hexanoyl-L-carnitine0.47260.285.1
33.Glutamine0.45147.184.1
34.C5-Valeryl-L-carnitine0.48246.285.1
35.C5-Valeryl-L-carnitine-d30.49249.185.1
36.C5-Isovaleryl-L-carnitine0.49246.185.1
37.C5-Isovaleryl-L-carnitine-d30.49249.285.1
38.2-Methylbutyryl-L-carnitine0.49246.285.1
39.C5:1-Tiglyl-L-carnitine0.50244.285.1
40.C4-Butyryl-L-carnitine0.51232.285.1
41.C4-Butyryl-L-carnitine-d30.51235.285.1
42.C4-Isobutyryl-L-carnitine0.51232.185.1
43.C4-Isobutyryl-L-carnitine-d30.51235.185.1
44.Citrulline0.51176.1113.1
45.Glutamic acid0.55148.183.9
46.C3-Propionyl-L-carnitine0.54218.185.1
47.C3-Propionyl-L-carnitine-d30.54221.285.1
48.C2-Acetyl-L-carnitine0.56204.185.1
49.C2-Acetyl-L-carnitine-d30.56207.185.1
50.Arginine0.66175.270.1
51.Ornithine0.69133.170.1
All analytes were present endogenously at varying concentrations in whole blood, except the deuterated internal standards.
image
LC_CF0759
ColumnRaptor HILIC-Si EXP guard cartridge column (cat.# 9310A0252)
Dimensions:5 mm x 2.1 mm ID
Particle Size:2.7 µm
Temp.:45 °C
Sample
Diluent:85:15 Acetonitrile:water (v/v)
Conc.: Endogenous levels
Inj. Vol.:2.0 µL
Mobile Phase
A:30 mM Ammonium formate in water
B:Acetonitrile
Time (min)Flow (mL/min)%A%B
0.000.51585
0.40.57030
0.410.51585
1.20.51585
DetectorMS/MS
Ion Source:Electrospray
Ion Mode:ESI+
InstrumentUHPLC
Notes50 µL of whole blood was spotted on to Whatman 903 neonatal protein saver cards, which were then dried for 1 hour at room temperature. A 3.0 mm disk (∼3.0 µL whole blood) was punched out of the dried spot and into a 2.0 mL Eppendorf tube. 200 µL of 85:15 acetonitrile:water (v/v) that was fortified with known concentrations of stable isotope-labeled internal standards was added, and then the sample was vortexed and incubated for 20 minutes at room temperature on a microplate shaker at a speed of 400 rpm. The sample was then centrifuged for 10 minutes at 4000 rpm, and 150 µL of the supernatant was filtered using a Thomson SINGLE StEP Nano filter vial (cat.# 25882) prior to LC-MS/MS analysis.
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Acylcarnitines

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Aminoacids

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Guard

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Screening

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Dried blood spot

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