ACE UHPLC and HPLC Columns
Prednisolone, Prednisone, Cortisol and Cortisone in Serum by LC-MS/MS
Column: ACE Excel 2 C18
Dimensions: 100 x 2.1 mm
Part Number: EXL -101-1002U
Mobile Phase: A: 4 mM ammonium acetate in H2O
B: 0.2% (v/v) formic acid in MeOH
Gradient: Time (mins) %B
0.00 30
0.25 30
3.90 70
6.00 70
6.01 95
7.00 95
7.01 100
Flow Rate: 0.2 mL/min
Injection: 50 μL
Temperature: 50 °C
Detection: Applied Biosystems 5000 MS/MS
APCI in positive ion mode
Preservatives (I)
140
120
100
80
60
40
20
144
1. Prednisolone
(m/z 361.5 147.1)
2. Cortisol
(m/z 363.5 121.3)
3. Prednisone
(m/z 359.4 147.1)
4. Cortisone
(m/z 361.5 163.3)
5. Prednisolone-d6 (IS)
(m/z 367.4 150.3)
6. Cortisol-d4 (IS)
(m/z 367.3 331.3)
2.0 4.0 6.0 8.0 10.0 12.0 2.0 4.0 6.0 8.0 10.0 12.0
2.0 2.0
4.0 4.0
6.0 6.0
2
8.0 8.0
10.0 10.0
12.0 12.0
5
2.0 2.0
4.0 4.0
Phthalic acid 4-Hydroxybenzoic acid Sorbic acid
2
Preservatives (II)
Methyl paraben Ethyl paraben
Propyl paraben
Response - MilliVolts
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Application #AN2690
Conditions Analytes
1
3
6.0 6.0
8.0 8.0
10.0 10.0
12.0 12.0
Reproduced with permission of Department of Pathology and Laboratory Medicine, Heartlands Hospital, Birmingham, UK
4
6
Prednisolone Cortisol
Cortisone
Prednisone
Prednisolone-d6 (IS) Cortisol-d4 (IS)
Time - Minutes Time - Minutes
Column: ACE Excel 1.7 C18
Dimensions: 50 x 3.0 mm
Part Number: EXL -171-0503U
Mobile Phase: 20 mM potassium phosphate
pH 2.5 in MeCN/H2O (30:70 v/v)
Flow Rate: 0.43 mL/min
Injection: 0.7 μL
Temperature: 20 °C
Detection: UV, 230 nm
Application #AN2230
Conditions Analytes
1. Phthalic acid
2. 4-Hydroxybenzoic acid
3. Sorbic acid
4. Methyl paraben
5. Ethyl paraben
6. Propyl paraben
1 2 3 4 5 6 7 8 9 min
mAU
0
1
3
4
5
6
Column: ACE 5 C18
Dimensions: 250 x 4.6 mm
Part Number: ACE-121-2546
Mobile Phase: MeCN/50 mM KH2PO4
pH 4.4 in H2O (40:60 v/v)
Flow Rate: 1 mL/min
Temperature: Ambient
Detection: UV, 230 nm
Application #AN3040
Conditions Analytes
1. Phthalic acid
2. p-Hydroxybenzoic acid
3. Benzoic acid
4. Sorbic acid
5. Methyl paraben
6. Ethyl paraben
7. Propyl paraben
1
2
3
4
5 6 7
Time - Minutes Time - Minutes