Page 43 - Application Notebook - Solution for Food Development

Application No.M271
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2. Analysis of Metabolites Present in Foods n Results
n Sample Taking the results from analysis, peak identification was
Next, metabolites present in foods were extracted from performed for compounds registered in the Smart
each sample, derivatized, and analyzed by GC-MS. Metabolites Database based on their quantitative ions,
We took 20 µL of each sample, added 60 µL of an reference ions, and retention indices. The numbers of
aqueous solution of ribitol (0.2 mg/mL) as an internal compounds identified are shown in Table 5. The 149
standard solution, and dried this mixture thoroughly in compounds detected by this analysis are also listed in
a centrifugal concentration device. To the dried residue Table 6.
was added 100 µL of a methoxyamine hydrochloride/ Principal Component Analysis (PCA) was performed for
pyridine solution (20 mg/mL), and this mixture was the 138 compounds detected in all samples. A score
shaken at 30 °C for 90 minutes. Subsequently, 50 µL of plot of this analysis is shown in Fig. 3. The three
N-Methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA) different sake types are clearly separated on the score
was added, and the mixture was shaken at 37 °C for plot. A loading plot of this analysis is shown in Fig. 4.
30 minutes. This sample was then added to a GC-MS Compounds characteristic to each sample were
vial and used for analysis. identified from these results.
The results suggest that performing a complete analysis
of metabolites and subsequent multivariate analysis of
identified compounds may be useful for food quality
n Analytical Conditions evaluation.
Table 4 Analytical Conditions for Analysis of Metabolites
Present in Foods
Triple quadrupole gas chromatograph mass spectrometer
: GCMS-TQ8040
Optional software : Smart Metabolites Database 6
Daiginjo-shu
4 Futsu-shu
GC Futsu-shu 3 Daiginjo-shu1
Column : BPX5 2 Futsu-shu 1 Daiginjo-shu 3
(30 m × 0.25 mm I.D., 0.25 µm) Futsu-shu 2 Daiginjo-shu 2
Carrier Gas : He t [2] 0
Control Mode : Linear velocity (39.0 cm/sec) -2 Junmai-shu 1
Injection Method : Split -4 Junmai-shu 3 Junmai-shu 2
Split Ratio : 30 Junmai-shu
Oven Temperature : From 60 °C (2 min) by (15 °C/min) to -6
330 °C (3 min)
-8
-15 -10 -5 0 5 10
MS (EI method)
Ion Source Temperature : 200 °C
Interface Temperature : 280 °C Fig. 3 Score Plot of the Analysis of Metabolites Present in Foods
Tuning Mode : Standard
Measurement Mode : MRM
Loop Time : 0.25 seconds

0.5 Galactose-5TMS Tehalose-8TMS
Table 5 Numbers of Compounds Detected by Mannose-meto-5TMS
Analysis of Metabolites Present in Foods 0.4
Futsu-shu Junmai-shu Daiginjo-shu 0.3 Maltose-meto-8TMS
Detected 147 140 149 0.2 Psicose-meto-5TMS
compounds 0.1
p [2] 0 Phosphoric acid-3TMS
-0.1
-0.2
Ornithine-4TMS
-0.3
-0.5 -0.4 -0.3 -0.2 -0.1 0 0.1 0.2 0.3 0.4
p[1]
Glucose-meto-5TMS

Fig. 4 Loading Plot of the Analysis of Metabolites Present in Foods