Department of Pharmaceutical Analysis, Karnataka College of Pharmacy, Thirumenahalli, Bangalore
In this research work, Extraction of HMF (5-hydroxymethylfurfural) from honey was done and the amount was estimated by using the reference standard of HMF on HPLC. Honey samples were received for testing from two companies and the reference standard was obtained from Sigma-Aldrich. Honey is a natural product which is produced by honey bees (Apis mellifera). The composition of honey changes depending on its floral, geographical and entomological sources. HMF concentration in fresh honey is mostly absent or is present in very low amounts, but its concentration increases during processing, upon storage for longer periods of time and upon heating/storing at higher temperatures. HMF contributes to sometimes fatal effects (mutagenic, chromosomal aberrations, genotoxic, organotoxic, carcinogenic, cytotoxicity towards mucous membranes, the skin, the eyes and the upper respiratory tract). Extraction of HMF from honey was carried out by making Carrez I and Carrez II solutions. An accurate and specific Reversed-Phase HPLC method was developed due to the urgent requirement of an analytical method to estimate the quantity of HMF in honey. A mobile phase with a combination of Acetonitrile and Water with 0.2% Formic Acid was used with a flow rate of 0.6 ml/minute and the separation was done on Phenomenex Non-Polar C18 Column with dimensions of 150 mm X 4.6 mm X 3 ?m at a wavelength of 282 nm. Analysis was carried out with a run time of 15 minutes. The Calibration curve method was adopted for the estimation of HMF in honey with R2 value of 0.999. These results were repeated two times in the laboratory; hence Repeatability Precision was performed to validate the results. The retention time of the HMF reference standard was 6.53 minutes and retention time of the samples (Sample 1 and Sample 2) was 6.63 minutes and the retention time variation of ± 0.1 minutes was observed which is acceptable and was within limits. The amount of HMF in honey in the samples were found to be within the limit of 40 mg/kg as per FSSAI. The amount of HMF present in Sample 1 and Sample 2 was found to be 2.20 mg/kg and 5.28 mg/kg respectively. HMF was extracted from honey and a new analytical HPLC method was developed. This new HPLC method developed would be economical, reproducible, and repeatable. The amount of HMF present in Honey samples was found to be 3.80 mg/kg and 6.50 mg/kg. The results were found to be within the limits (40 mg/kg) as specified by FSSAI regulatory guidelines and Sample 2 was found to be 2.20 mg/kg and 5.28 mg/kg respectively. HMF was extracted from honey and a new analytical HPLC method was developed. This new HPLC method developed would be economical, reproducible, and repeatable. The amount of HMF present in Honey samples was found to be 3.80 mg/kg and 6.50 mg/kg. The results were found to be within the limits (40 mg/kg) as specified by FSSAI regulatory guidelines.
: P. Singh, C. Sreedhar, Quantification of a Carcinogenic and Cytotoxic compound: HMF(5-hydroxymethyl-2-furfural) using HPLC(High-Performance Liquid Chromatography), Int. J. of Pharm. Sci., 2024, Vol 2, Issue 1, 624-632. https://doi.org/10.5281/zenodo.10568997
10.5281/zenodo.10568997
