LC profiles for 1–5% combined working standard of glucose, fructose and sucrose are represented in while sample profile is obvious in . Differential Refractometer (KNAUER) ICI – Comp (Range 16.3) was used in this analysis. The refractive index (RI) is bulk property and a virtually universal detector based on density. However, this detector does lack its sensitivity. In LC, a normal phase chromatography is a separation mode which runs on unbonded, anhydrous silica (polar) using a nonpolar mobile phase. Generally, in such a case of LC, properties of mobile phase such as pH and concentration have vital roles in controlling the retention time, therefore, a different mode of gradient was applied to allow the strongly retained analyte to be eluted quickly. In this sugar analysis, percentage ratio of acetonitrile and water played this role. Usually, decreasing the particle size of the column packing material and reducing the column length produces better peaks and reduces retention time.
LC profile for 1% combined working standard of glucose, fructose and sucrose.
LC profile for 2% combined working standard of glucose, fructose and sucrose.
LC profile for 3% combined working standard of glucose, fructose and sucrose.
LC profile for 5% combined working standard of glucose, fructose and sucrose.
LC profile for honey sample (duplicate experiment).
According to slope values obtained after linear regression analysis of the data (), it was found that fructose and glucose was found as 1.605 ± 0.021 and 1.08 ± 0.085 (SD)% in the sample profile of honey while after multiplication with the dilution factor it becomes 40% and 27% respectively. A setup of integration parameters such as threshold and peak width in this current study was important. This is critical because it allows the process to seek the required peaks within the chart size. Furthermore, it allows the ability to achieve effectively the study of comparable obvious heights. If this setup is inappropriate, it can lead to number of consequences. These are creation of unnecessary peaks or very short peaks or abnormal high height of the peaks, to be observed, that goes out of the range. Therefore, appropriate set up was necessary before the running of the sample or standard on LC.
Plot for area of working standard sugars versus concentration.
According to the literature, honey has always been regarded as a food which is advantageous for one’s health and as a product that has healing qualities. For this reason, it is necessary to protect consumers from the fraudulent mislabeling of inferior honeys, therefore, Feás et al. (2010)
characterized artisanal honey is produced on the Northwest of Portugal by melissopalynological and physico–chemical analysis. A method has been also developed and validated for the simultaneous analysis of different veterinary drug residues (macrolides, tetracyclines, quinolones, and sulfonamides) in honey by Vidal et al. (2009)
. Overall this study is composed of a simple protocol for the analysis of monosaccharide in the honey, which can be a guideline for students and junior researchers to demonstrate normal phase partition LC, operation and application of the differential refractometer detector.