Page 11 - LifeScience Solutions for Stem Cell Analysis

Metabolomics by LC-MS/MS
LC-MS/MS
Data

Hydrophilic metabolites (96 components) including amino Table 1 shows MRM transitions of the typical of
acids, phosphorylated sugars (compounds in glycolysis), compounds. Using yeast extracts as a biological model
organic acids (compounds in the TCA cycle), coenzymes source, 75 components were detected successfully at
such as NADH, and nucleotides such as ATP were analyzed concentration range between 0.4 – 100,000 nM (Fig. 2). iPS Cell Differentiation Proteomic Analysis of Human
successfully within 15 minutes using the LCMS-8040 (Fig. 1).

(x100,000) Table 1 Typical compounds
6.0
5.0 # Compound MRM transition
4.0 1 Arginine 173.1>131.2
3.0 2 Histidine 154>93.15
2.0 3 4-Aminobutyrate 102>84 of Human ES Cells Comparative Metabolomics
1.0 4 Serine 104>74.15
0.0 5 Asparagine 131>113.15
0.0 2.5 5.0 7.5 10.0 min
6 Glutamine 145>127.05
Fig. 1 Typical chromatograms of 96 standard compounds
7 Hydroxyproline 130>84.15
(x1,000,000)
2.5 8 Homoserine 118>100
9 Threonine 118>74.05
2.0
10 Leucine 130.1>84
1.5 a Super-functional Liver Chip A Micro-fluidic Approach using
11 Ribitol 151>89.1
1.0 12 Trehalose 341>89
13 Proline 114>68.1
0.5
14 Cytidine 242>109.15
0.0
0.0 2.5 5.0 7.5 10.0 min 15 Methionine 148>47.05
Fig. 2 Typical chromatograms of yeast extract

Linearity of G6P (Glucose-6-phosphate, a glycolysis by LC-MS/MS Metabolomics
compound) was tested through serial dilution of a yeast
extract to confirm the linear range (Fig. 3). This result also
demonstrates that no ion suppression was present in the
analytical system.

Fig. 3 Linearity of a yeast extract dilution series by MALDI Tissue Imaging

UHPLC LC-MS
Instrument : Nexera series UHPLC Instrument : LCMS-8040 triple-quadrupole mass spectrometer
Mobile phase : A: 10 mM tri-butyl ammonium and 15 mM acetic acid in Ionization : ESI negative mode
water/methanol (97/3) DL temp. : 250°C HB temp: 400°C
: B: methanol Drying gas : 10 L/min Nebulizing gas: 2 L/min
Flow rate : 0.3 mL/min
Time program : 0%B (0-0.5 min) - 25%B (7.5 min) -90%B (11-11.5 min) -
0%B (11.6-15 min)
Analytical column : L-Column ODSII (2 mm I.D. × 150 mmL., 3 µm)
Oven temp. : 40°C Spectrometers Shimadzu’s Mass
Injection vol. : 3 µL

11