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Preparation of Fructo Oligosaccharide Sample

Application Introduction

The sample is extracted from the material system containing fructo oligosaccharide. The oligosaccharides including kestose, nystose, 1F-fructofuranosylnystose, etc.

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Introduction

The sample is extracted from the material system containing fructo oligosaccharide. The oligosaccharides including kestose, nystose, 1F-fructofuranosylnystose, etc. shall be separated from the sample. This experiment establishes a preparative purification method to prepare multiple types of fructo oligosaccharide monomers.

Table 1 Fructo Oligosaccharide Sample Information

Sample   name

English   name

Structural   formula

Molecular   formula

Molecular   weight

CAS   No.

Fructo oligosaccharide

Fructo   oligosaccharide

Unkown

Unkown

Unkown

Unkown

Experimental

Instruments, reagents and materials

Cheetah HP100 Prep HPLC and evaporative light scattering detector

Chromatographic solvent: acetonitrile; water: deionized water

HPLC columns: Venusil® HILIC (10 µm, 100 Å, 300 g)

Sample pretreatment

The sample is white powder. Dissolve 3 g of sample ultrasonically with 5 mL of water. Filter the sample with a 0.22 µm membrane, and then perform the sample loading.

Preparation conditions

Chromatographic columns: Venusil® HILIC (10 µm, 100 Å, 300 g) packed in DAC (inner diameter: 50 mm);

Mobile phases: A: water, B: acetonitrile;

Detector: ELSD 70 °C

Flow rate: 80 mL/min

Sample size: 5 mL

Gradient:

Table 2 Gradient Elution Conditions of Liquid Chromatography

Time/min

Flow rate/(mL/min)

A/%

B/%

0.0

80.0

23.0

77.0

180.0

80.0

23.0

77.0

Detection conditions

Chromatographic columns: Venusil® HILIC (5 µm, 100 Å, 4.6 × 250 mm);

Mobile phases: A: water, B: acetonitrile;

Detector: ELSD 60 °C

Flow rate: 1.0 mL/min

Sample size: 5 µL

Column temperature: 30 °C

Gradient:

Table 3 Gradient Elution Conditions of Liquid Chromatography

Time/min

Flow rate/(mL/min)

A/%

B/%

0.0

1.0

10.0

90.0

20.0

1.0

50.0

50.0

30.0

1.0

50.0

50.0

Results and Discussion

In this experiment, fructo oligosaccharide sample is prepared with Venusil® HILIC (10 µm, 100 Å, 300 g) packing. As shown in Table 2 and Figure 1, the sample prepared shows a good peak shape after going through the gradient elution in the mobile phases including water and acetonitrile. Besides, the purity of the product prepared is conforming to the requirement of customer.

Figure 1 Preparation Chromatogram

Figure 2 Detection Chromatogram

Figure 3 Empty Chromatogram

Figure 4 Detection Chromatogram of Fraction at 15min

Figure 5 Detection Chromatogram of Fraction at 22min

Figure 6 Detection Chromatogram of Fraction at 34min

Figure 7 Detection Chromatogram of Fraction during 40~47min

Figure 8 Detection Chromatogram of Fraction during 53~64min

Figure 9 Detection Chromatogram of Fraction during 73~85min

Figure 10 Detection Chromatogram of Fraction during 105~117min

Figure 11 Detection Chromatogram of Fraction during 140~160min

Conclusion

In this experiment, fructo oligosaccharides are prepared with Venusil HILIC (10 µm, 100 Å, 300 g) chromatographic columns. Results show that Venusil HILIC packing has a good effect on this sample in separation, and the purity of product prepared is conforming to the requirement of customer.


  • Venusil HILIC

    Venusil HILIC stationary phase is made from ultra pure spherical silica particles bonded with a neutral hydrophilic amide group. It can be used as normal phase, reversed phase or hydrophilic interaction HPLC. The Venusil HILIC column is a suitable replacement for NH2 and silica columns that are currently marketed as HILIC.

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