AI-64310-CRFT

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2
Table 2. LC gradient
Mass Spectrometry
The LC system was connected to
Acquisition
time ranges were used for each analyte and the following
MS conditions were used:
Group Time(min)
Flow Rate
(mL/min)
A
(%)
B
(%)
1
0.0 0.8 100 0
0.1 0.8 100 0
0.5 0.8 60 40
2.5 0.8 60 40
3.0 1.0 0 100
4.0 1.0 0 100
4.1 0.8 100 0
2
0.0 1.0 100 0
0.1 1.0 100 0
1.0 1.0 0 100
3.0 1.0 0 100
3.1 1.0 100 0
3
0.0 1.0 100 0
0.1 1.0 100 0
1.0 1.0 0 100
3.0 1.0 0 100
3.1 1.0 100 0
Liquid Chromatography
Liquid chromatography analysis was performed using a
The LC
conditions were as follows:
LC column Provided with the kit
Mobile phase A Provided with the kit
Mobile phase B Provided with the kit
Injection volume Group 1 – 20 µL
Group 2 – 100 µL
Group 3 – 30 µL
LC gradient See Table 2
Source type Heated electrospray ionization (HESI)
Vaporizer temp 350 °C
Capillary temp 350 °C
Spray voltage 3500 V
Sheath gas 70 AU
Sweep gas 0 AU
Auxiliary gas 40 AU
Data acquisition mode Selected-reaction monitoring (SRM)
Chrom filter peak width 5.0 s
Collision gas pressure 1.5 mTorr
Cycle time 0.300 s
Q1 mass resolution (FWMH) 0.7
Q3 mass resolution (FWMH) 0.7
SRM settings See Tables 3, 4 and 5
Group Analyte CAL 1 CAL 2 CAL 3 CTRL1 CTRL2
1
Carbamazepine 1.62 8.77 15.1 3.25 10.6
Oxcarbazepine 0.14 1.89 3.64 0.46 2.75
Carbamazepine diol 0.16 5.77 11.5 1.11 8.21
Carbamazepine-10,11-epoxide 0.16 5.61 11.0 1.07 8.1
10-hydroxycarbamazepine 3.83 26.6 48.3 8.48 36.1
Felbamate 14.3 70.8 130 27.2 92.9
Lacosamide 0.78 6.13 11.4 1.91 8.5
Levetiracetam 4.90 28.5 84.0 16.0 62.8
Rufinamide 3.98 21.0 35.3 7.54 28.0
Theophylline 6.11 15.9 24.6 9.78 18.9
2
Phenytoin 3.49 13.3 23.3 5.86 16.9
Primidone 3.07 10.7 18.3 5.11 13.6
Phenylethylmalonamide (PEMA) 1.07 6.68 12.7 2.21 9.03
Stiripentol 2.62 14.3 27.3 5.01 20.3
3
Phenobarbital 7.38 34.2 60.1 13.8 44.7
Valproic Acid 31.6 79.1 125 47.2 97.7
Zonisamide 3.97 27.4 50.4 9.04 36.8
Table 1. Concentrations (ng/mL) of calibrators and controls