ACE UHPLC and HPLC Columns
Conditions
1. Ethyl glucuronide
Ethyl glucuronide (EtG)
LOQ in H2O 300 fg on-column EtG 100 pg/mL
2.25
2.00
1.75
1.50
1.25
1.00
0.75
0.50
0.25
m/z 221.2 75.0
m/z 221.2 84.9
0.0 0.5 1.0 1.5 2.0 2.5 3.0
(x100,000)
1:EtG 221.20>84.90(-) CE: 15.0
1:EtG 221.20>75.00(-) CE: 15.0
1:EtG 221.20>57.00(-) CE: 22.0
1:EtG 221.20>113.00(-) CE: 14.0
Ethyl glucuronide-d5 (EtG-d5) EtG-d5 100 pg/mL
2.50
2.25
2.00
1.75
1.50
1.25
1.00
0.75
0.50
0.25
0.00
2.50
2.25
2.00
1.75
1.50
1.25
1.00
0.75
0.50
0.25
2:EtG-d5 226.30>85.00(-) CE: 16.0
2:EtG-d5 226.30>74.90(-) CE: 16.0
2:EtG-d5 225.80>57.10(-) CE: 24.0
Reproduced with permission of Shimadzu Europa GmbH. Analysed on Shimadzu Nexera X2
74
9
pH 3
2.50
2.25
2.00
1.75
1.50
1.25
1.00
0.75
0.50
0.25
2 3 4 5 6 min
1
3
2
4
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7 8
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pH 10.7
Ethyl Glucuronide in Water by LC-MS/MS
Exploiting Selectivity by Adjusting pH
Conditions
Application #AN1100
Application #AN2440
Analyte
Analytes
1. Nizatidine
2. Salbutamol
3. Amiloride
4. N-Acetylprocainamide
5. Quinoxaline
6. Methyl paraben
7. p-Cresol
8. Reserpine
9. Piperine
10. Toluene
11. Felodipine
Column: ACE Excel 2 C18-PFP
Dimensions: 100 x 2.1 mm
Part Number: EXL -1010-1002U
Mobile Phase: A: 0.05% formic acid in H2O
B: MeOH
Gradient: Time (mins) %B
0.00 5
4.00 70
6.00 95
7.00 95
7.01 5
Flow Rate: 0.4 mL/min
Injection: 3 μL
Temperature: 40 °C
Detection: Shimadzu LCMS-8050
ESI voltage: -3 kV
Desolvation line: 250 °C
Interface heater: 380 °C
Nebulizing gas: 3 L/min
Heat block: 400 °C
Column: ACE Excel 2 SuperC18
Dimensions: 50 x 2.1 mm
Part Number: EXL -1011-0502U
Mobile Phase: A1: 10 mM ammonium
formate pH 3.0 in H2O
A2: 0.1% ammonia pH 10.7 in H2O
B1: 10 mM ammonium formate pH
3.0 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
0.0 3
7.0 100
8.0 100
8.5 3
12.5 3
Flow Rate: 0.42 mL/min
Injection: 2 μL
Temperature: 40 °C
Detection: UV, 254 nm
Time - Minutes
pH 3
pH 10.7
0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 min
0.00
(x100,000)
2:EtG-d5 226.10>112.90(-) CE: 14.0
0.5 1.0 1.5 2.0 2.5 3.0
0.00
(x100)
2:226.30>74.90(-)
2:226.30>85.00(-)
0.00
(x100)
1:221.20>75.00(-)
1:221.20>84.90(-)
0.00 0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 min
m/z 226.3 85.0
m/z 226.3 74.9
m/z 225.8 57.1
m/z 226.1 112.9
m/z 226.3 74.9
m/z 226.3 85.0
m/z 221.2 84.9
m/z 221.2 75.0
m/z 221.2 57.0
m/z 221.2 113.0
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