ACE UHPLC and HPLC Columns
Metabolomic Analysis of Extracted Jurkat T Cells by LC-HRMS
Column: ACE Excel 2 C18-PFP
Dimensions: 100 x 2.1 mm
Part Number: EXL -1010-1002U
Mobile Phase: A: 0.1% formic acid in H2O
B: 0.1% formic acid in MeCN
Gradient: Time (mins) %B
0 0
1 0
11 65
13 65
18 95
20 95
Flow Rate: 0.35 mL/min
Injection: 5 μL
Temperature: 35 °C
Detection: Thermo Scientific Q Exactive Orbitrap MS
Heated electrospray ionisation in positive mode
Spray Voltage: 3.3 kV
Capillary Temperature: 300 °C
Heater Temperature: 350 °C
Mass Scan Range: m/z 70-1000
Resolution: 70,000
1. Caffeine-d3 (IS)
2. Tryptophan-d3 (IS)
3. 1,3,7-Octanetriol
4. Pimelylcarnitine
40
35
30
25
20
15
10
5
Metabolomic Biomarkers in Ethylmalonic Encephalopathy
1,3,7-Octanetriol Pimelylcarnitine
18
16
14
12
10
8
6
Time - Minutes
4
5
6 7
2 4 6 8 10 12 14 16
Time - Minutes
3
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Column: ACE 3 C18-PFP
Dimensions: 150 x 2.1 mm
Part Number: ACE-1110-1502
Mobile Phase: A: 0.1% formic acid in H2O
B: 0.1% formic acid in MeCN
Detection: Sciex API 4000 triple quad MS
ESI in negative ion mode
Sample: Urine sample from patient with ethylmalonic
encephalopathy is filtered and extracted with
ice-cold methanol, evaporated to dryness and
reconstituted in 0.1% formic acid in water
Please contact info@ace-hplc.com for additional information on
the chromatographic conditions used for this analysis.
Application #AN3980
Application #AN4130
Conditions
Conditions
Analytes
Analytes
1. Ethylmalonic acid
(m/z 131 87)
2. Methylsuccinic acid
(m/z 131 87)
3. Adipic acid
(m/z 145 83)
4. Isobutyrylcarnitine
(m/z 232 85)
5. Butyrylcarnitine
(m/z 232 85)
6. 2-Methylbutyrylcarnitine
(m/z 246 85)
7. Isovalerylcarnitine
(m/z 246 85)
8. Valerylcarnitine
(m/z 246 85)
1 2
3 4
0 2 4 6 8 10 12 14 16
0
6 7 8 9 10 11 12
4
x108
2
3 4
1
Retention Time mins
Intensity
x108
TIC overlay for Jurkat T-lymphocyte cells rinsed with
either 0.3% ammonium formate (darker line) or 0.3%
ammonium acetate.
Ulmer CZ, Yost RA, Chen J, Mathews CE, Garrett TJ. Liquid-Chromatography-Mass Spectrometry Metabolic and Lipidomic Sample Preparation Workflow for
Suspension-Cultured Mammalian Cells using Jurkat T Lymphocyte Cells, J. Proteomics Bioinform, (2015), 8(6), 126-132. doi:10.4172/jpb.1000360
2
Time - Minutes
Reproduced with permission of UCSD Biochemical Genetics, Department of Pediatrics, University of California San Diego, California, USA
8
1
4 8 12 16 20 24 28 32 intensity
intensity
Negative ion mode Positive ion mode