Honey is a natural food primarily composed of monosaccharides such as glucose and fructose, along with vitamins, amino acids, polyphenols, and minerals. It may also contain disaccharides like sucrose and its isomer, turanose. In this application, glucose, fructose, sucrose, and turanose were analyzed using HILICpak VG-25 4D, a polymer-based amino column. In the acacia honey sample used for this analysis, a peak for turanose was observed, but sucrose peak was not detected. 
In general sugar analysis using HILIC mode, separating sucrose and turanose is challenging. However, by using VG-25 4D and adding methanol to the eluent, made it feasible to separate sucrose and turanose.

Sample Preparation

*In accordance with the "Operating Procedure for testing of honey s" by JAPAN Honey Fair Trade Council.
Standard Solution

1. 1.Dispense 25 mL of methanol into a 100 mL volumetric flask.
2. 2.Weigh 1.5 g of glucose, 2.0 g of fructose, 0.25 g of sucrose, and 0.25 g of turanose into a 100 mL beaker and dissolve them in approximately 40 mL of ultrapure water.
3. 3.Transfer the entire solution from step 2 into the volumetric flask from step 1 and make up to 100 mL with ultrapure water.
4. 4.Filter the solution through a 0.45-µm membrane filter to prepare the HPLC sample solution.

1. 1.Accurately weigh 5 g of honey into a beaker and dissolve it in 40 mL of ultrapure water.
2. 2.Dispense 25 mL of methanol into a 100 mL volumetric flask, transfer the entire honey solution from step 1, and make up to 100 mL with ultrapure water.
3. 3.Mix thoroughly and filter the solution through a 0.45-µm membrane filter and use it as a sample.

Sample: 10 µL each
Stadard solution

1. 1.Fructose 2 %
2. 2.Glucose 1.5 %
3. 3.Sucrose 0.25 %
4. 4.Turanose 0.25 %

Sample solution (Acacia honey)

1. 1.Fructose
2. 2.Glucose
3. 4.Turanose