ACE UHPLC and HPLC Columns
rT3 (Reversed tri-iodothyronine)
mAU
mAU
8
6
4
2
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Thyroid Hormones by LC-MS/MS (I)
Application #AN2170
Conditions Analytes
1. T3 (Tri-iodothyronine)
2. rT3 (Reversed tri-iodothyronine)
3. T4 (Thyroxine)
Column: ACE Excel 2 C18-AR
Dimensions: 100 x 2.1 mm
Part Number: EXL -109-1002U
Mobile Phase: A: 2 mM ammonium acetate, 0.1%
formic acid in H2O
B: 2 mM ammonium acetate, 0.1%
formic acid in MeOH
Gradient: Time (mins) %B
0.0 60
3.0 77
3.1 60
Flow Rate: 0.4 mL/min
Injection: 10 μL
Temperature: 40 °C
Detection: XEVO TQS triple quad MS
Desolvation temperature: 500 °C
Ion source temperature: 150 °C
Positive mode ESI, MRM
Sample: Serum samples extracted using Biotage
EVOLUTE EXPRESS AX methodology
1
3
2
TIC
1
3
2
TIC
Analyte Q1 (Da) Q3 (Da)
T3 651.8 605.8
(651.8) (507.8)
(651.8) (478.9)
rT3 651.8 605.8
(651.8) (507.8)
(651.8) (478.9)
T3/rT3-d6 I.S. 657.8 611.8
T4 777.7 731.7
(777.7) (351)
(777.7) (633.8)
Reproduced with permission of Biotage GB Ltd, UK
T3 (Tri-iodothyronine)
T4 (Thyroxine)
Time - Minutes
Tocopherols Mixture Separation
Application #AN3390
Conditions
Column: ACE Excel 3 C18-PFP
Dimensions: 150 x 4.6 mm
Part Number: EXL -1110-1546U
Mobile Phase: A: 0.1% H3PO4/MeCN (1:3 v/v)
B: MeCN
Gradient: Time (mins) %B
0.00 0
0.10 0
0.11 80
8.00 80
8.01 100
12.00 100
12.01 0
14.00 0
Flow Rate: 1.2 mL/min
Injection: 10 μL
Temperature: 40 °C
Detection: UV, 285 nm
1. γ-Tocopherol
2. β-Tocopherol
3. δ-Tocopherol
4. α-Tocopherol
5. α-Tocopherol acetate
8
6
4
2
0
9
mins
1 2
3 4
5
5 6 7 8 9
0
2
3 4
5
1
Analytes
γ-Tocopherol β-Tocopherol
δ-Tocopherol α-Tocopherol
α-Tocopherol acetate
Time - Minutes
Reproduced with permission of WIREC-WESSLING International Research and Educational Centre, Budapest, Hungary