Page 11 - Application Notebook - Solution for Food Development

Application No.L474
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n Quantitative Analysis of Oligosaccharides Included in Sake
We used the ELSD to conduct quantitative analysis of Sake 1 mL Water 1 mL
glucose, isomaltose and panose that are present in ↓ Acetonitrile 3 mL
sake. Pretreatment of the sake sample was performed Mixing
according to the steps shown in Fig. 3. The calibration ↓
curves that were generated based on standard samples Filtration (0.2 µm membrane filter)
analyzed using the analytical conditions shown in Table ↓
2 are shown in Fig. 4. Because the ELSD is not a
spectroscopic detector, it does not obey Beer's law with HPLC 5 µL inj.
a linear correlation between absorbance and
concentration. Instead, a log-log plot of peak area and Fig. 3 Pretreatment of Sake
analyte quantity produces a linear response. Within the
concentration range including 100, 200, 400, 1000,
and 2000 mg/L (for glucose, 200, 400, 800, 2000,
2
4000 mg/L), excellent linearity was obtained with R
greater than 0.999. Fig. 5 shows the chromatogram of
sake obtained using the conditions shown in Table 2.
The quantitation results calculated using the calibration
curves generated using the standard samples are shown
in Table 3.

17 14 15
16 R² = 0.9991 13 R² = 0.9996 R² = 0.9995
14
15 12 13
Ln (Area) 14 Ln (Area) 11 Ln (Area) 12
10
13
12 9 11
Glucose Isomaltose Panose
11 8 10
10 7 9
5 6 7 8 9 4 5 6 7 8 4.5 5.5 6.5 7.5
Ln (mg/L) Ln (mg/L) Ln (mg/L)

Fig. 4 Calibration Curves (Each injection 5 µL)

Table 3 Quantitation Results and Repeatability (n = 3)
mV of Oligosaccharides in Japanese Sake
450 Area %RSD Conc.
1 n Peaks
400 1. Glucose Glucose 432×10 4 0.6 13110 mg/L
2. Isomaltose Isomaltose 228×10 3 2.1 5130 mg/L
350 3. Panose
Panose 909×10 2 5.9 1410 mg/L
300
250 Note: The listed concentrations are the values obtained
after converting the results obtained from the 5 µL
200
measurement samples to the undiluted source
150 solution.
100
3
50 2
0
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 min
Fig. 5 Chromatogram of Sake

First Edition: Nov. 2014

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