ACE UHPLC and HPLC Columns
Pharmaceutically Relevant Mixture (II) – Different Selectivity Using pH
11
ppHH 1 100.7.7
1 2
3
4
5
6 7 8
9
10
Phenelzine in Human Plasma by LC-MS/MS
Spiked human plasma
Time - Minutes Time - Minutes
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Conditions
Column: ACE Ultracore 2.5 SuperPhenylHexyl
Dimensions: 50 x 2.1 mm
Part Number: CORE-25B-0502U
Mobile Phase: A1: 10 mM ammonium formate pH 3
in H2O
A2: 0.1% ammonia pH 10.7 in H2O
B1: 10 mM ammonium formate pH 3
in MeCN/H2O (90:10 v/v)
B2: 0.1% ammonia pH 10.7 in
MeCN/H2O (90:10 v/v)
Gradient: Time (mins) %B
0 3
5 100
6 100
Flow Rate: 0.6 mL/min
Temperature: 40 °C
Detection: UV, 254 nm
Application #AN1300
Analytes
1. Benzamide
2. Caffeine
3. Procainamide
4. N-Acetylprocainamide
5. Propiophenone
6. Toluene
7. Remacemide
8. Ethylbenzene
9. Carvedilol
10. Nortriptyline
11. Clomipramine
3
7
10
1 2
5
pH 3
3
1
4
2
7
5
9,6
10
11,8
pH 3
6 7
10
2
8
1
11
9
5
3
1 2 2 3 3 4 4 5 5 6
6 min
Time - Minutes
2
Column: ACE 5 C18
Dimensions: 100 x 4.6 mm
Part Number: ACE-121-1046
Mobile Phase: 10 mM ammonium acetate in
H2O, pH 4.0/MeOH (20:80 v/v)
Flow Rate: 1 mL/min with 70% split flow
into MS
Injection: 10 μL
Temperature: 45 °C
Detection: AB Sciex API-4000 MS
MRM using ESI in positive ion mode
TurboIonSpray Interface Temperature: 600 °C
IonSpray Voltage: 5500 V
Sample: Phenelzine derivatised with
pentafluorobenzaldehyde, followed by SPE
extraction of derivative and hydroxyzine
(I.S.) from 200 μL human plasma.
Application #AN4200
Conditions Analytes
1. Phenelzine derivative
(m/z 305.1 105.1)
20.2 ng/mL
2. Hydroxyzine
(m/z 375.3 201.1)
25 ng/mL
Phenelzine Phenelzine derivative Hydroxyzine
1
Intensity, cps
Reproduced with permission of School of Pharmacy, Texas Tech University Health Sciences Center, Amarillo, Texas, USA