Page 46 - Application Notebook - Solution for Food Development

Application No.C132
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n LC/MS/MS Analysis
Due to the difficulty of analyzing the chemical automatically changing between the three analytical
properties of primary and secondary metabolites under methods (Fig. 3). Although we used four mobile phases
the same chromatographic conditions, three and three columns for this experiment, the system itself
chromatographic conditions were chosen that differed can accommodate up to eight mobile phases and six
in mobile phase and column type. We used the Nexera columns. The analytical conditions used are shown in
method scouting system (comprehensive method Table 1.
search system) to perform measurements while

Pump A
A
B
C

1 2
1: water Auto Injector
2: 0.1 % formic acid-water LCMS-8050

Column Oven
Pump B
Column
A: Discovery HS F5-3 (2.1 × 150 mm, 3 µm)
B: Inertsil ODS-4 (2.1 × 50 mm, 2 µm)
1 2 C: Asahipak NH2P-50 2D (2.0 × 150 mm, 5 µm)

1: Acetonitrile
2: Acetonitrile / 2-propanol = 2/1

Fig. 3 System Configuration

Table 1 Analytical Conditions
Conditions Condition 1 Condition 2 Condition 3
Instrument UHPLC Nexera system (Shimadzu)
Organic acids Carotenoids Sugars
Target Compounds Amino acids
Flavonoids

A: Discovery HS F5-3 B: Inertsil ODS-4 C: Asahipak NH2P-50 2D
Column (150 mm L. × 2.1 mm I.D., 3 µm) (50 mm L. × 2.1 mm I.D., 2 µm) (150 mm L. × 2.0 mm I.D., 5 µm)
Sigma-aldrich GL-science Shodex
Column Oven Temp. 40 °C
HPLC
Mobile Phase A 2: 0.1 % Formic acid-Water 1: Water 1: Water
Mobile Phase B 1: Acetonitrile 2: Acetonitrile / 2-Propanol = 2/1 1: Acetonitrile
Flowrate 0.25 mL/min 0.4 mL/min 0.4 mL/min
0 %B (0-2 min.) ˠ 95 % (10-13 min.) 60 %B (0 min.) ˠ 100 % (5-8 min.) 65 %B (0-8 min.) ˠ 30 % (8-11 min.)
Time Program ˠ 0 % (13.01-16 min.) ˠ 60 % (8.01-10 min.) ˠ 65 % (11.01-15 min.)
Measurement Time 16 min 10 min 15 min
Total Run Time 41 min
Injection Volume 2 µL
Instrument LCMS-8050
MS Ionization ESI (+ / −)
Mode MRM