ACE UHPLC and HPLC Columns
Vitamin D2/D3
Conditions Analytes
Column: ACE Excel 2 C18-Amide
Dimensions: 50 x 3.0 mm
Part Number: EXL -1012-0503U
Mobile Phase: 100% MeCN
Flow Rate: 0.43 mL/min
Injection: 2 μL
Temperature: 20 °C
Detection: UV, 265 nm
Ergocalciferol (D2)
Cholecalciferol (D3)
5 6 2
1
Vi25-Hydroxy Vitamin D in Serum by LC-MS/MS
178
Application #AN1840
1. Ergocalciferol (D2)
2. Cholecalciferol (D3)
1
2
1 2 3 4 5 6 7 8
min
100
%
0
1a
2.00 2.10 2.20 2.30 2.40 2.50 2.60 2.70 2.80 2.90 3.00
100
%
0
1b
2.00 2.10 2.20 2.30 2.40 2.50 2.60 2.70 2.80 2.90 3.00
100
%
0
2
2.00 2.10 2.20 2.30 2.40 2.50 2.60 2.70 2.80 2.90 3.00
100
%
0
3a
2.00 2.10 2.20 2.30 2.40 2.50 2.60 2.70 2.80 2.90 3.00
100
%
0
1a. 25-OH Vitamin D2
(m/z 395.5 269.5)
1b. 25-OH Vitamin D2
(m/z 395.5 119.2)
2. d6-25-OH Vitamin D3 (IS)
(m/z 389.6 263.5)
3a. 25-OH Vitamin D3
(m/z 383.5 257.5)
3b. 25-OH Vitamin D3
(m/z 383.5 107.2)
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Application #AN2390
Conditions Analytes
Column: ACE Excel 2 C18-PFP
Dimensions: 100 x 2.1 mm
Part Number: EXL -1010-1002U
Mobile Phase: A: 2 mM ammonium acetate,
0.1% formic acid in H2O
B: 0.1% formic acid in MeOH
Gradient: Time (mins) %B
0.0 75
3.0 100
4.0 100
Flow Rate: 0.4 mL/min
Injection: 15 μL
Temperature: 40 °C
Detection: Quattro Premier XE triple quad MS
MRM positive ESI mode
Desolvation temperature: 450 °C
Ion source temperature: 150 °C
3b
25-OH Vitamin D2 25-OH Vitamin D3 2.00 2.10 2.20 2.30 2.40 2.50 2.60 2.70 2.80 2.90 3.00
Reproduced with permission of Biotage GB Ltd
Time - Minutes
Time - Minutes
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