HPLC Assay for the Determination of Ascorbic Acid in Honey Samples

Abstract

Determination of ascorbic acid in honey samples was performed by high performance liquid chromatography (HPLC) with UV detection at 254 nm. The separation was performed using a C-18-ODS column with a mobile phase consisting of a mixture of 15% methanol and 85% water, adjusted to pH 2.5 with metaphosphoric acid. The mobile phase was pumped at a flow rate of 0.9 mL/min. The retention time for ascorbic acid was found to be 5.1 minutes. The method is simple, fast, sensitive, and reproducible. The recovery of ascorbic acid is over 90% by the standard addition method.

Vitamin C and Sugar Levels as Simple Markers for Discriminating Spanish Honey Sources

“…Figure 1A shows the chromatogram of a vitamin C standard injected into the chromatographic system with a 0.01% H 2 SO 4 mobile phase, pH = 2.5, and T = 25 °C. The retention time for vitamin C was 2.7 min, this time was lower than the one obtained in octadecyl silane column C18 (5 μm) with other mobile phases such as methanol: H 2 O (15:85% v/v) in metaphosphoric acid at pH 2.5 (5.1 min) (Castro and others 2001) and H 2 SO 4 at pH 2.2 (9 min) (Vázquez‐Oderiz and others 1994).…”

“…Recently, vitamin C has been determined by reverse phase high‐performance liquid chromatography (RP‐HPLC) in isocratic mode, with a mobile phase of methanol/water at pH 2.5. This method is fast, it does not required sample treatment and the retention time for vitamin C is about 5 min (Castro and others 2001).…”

Voltammetric determination of ascorbic acid with zinc oxide modified glassy carbon electrode

“…According to Tauc, the dependence of the absorption coefficient α on the photon energy hυ for near-edge optical absorption in semiconductors takes the form (2):(αhυ) 1/m = k (hυ-Eg)(2)Where Eg is the optical band gap, k is a constant and m can take the values of 1/2 in the case of an allowed direct energy gap transition and 3/2 for a forbidden direct energy gap transition[30,31]. In order to determine the optical band gap of ZnO nanorods taking m = 1/2, (αhυ)2…”

CHAPTER 13. Assay of B Vitamins and other Water-soluble Vitamins in Honey

“…Consequently, the concentration data reported in the literature are spread over four orders of magnitude, going from less than 0.1 mg/kg to values very close to 3 g/kg!!! A number of research groups have reported concentrations of ascorbic acid in honey between 5.4 and 54 mg/kg (Abu-Tarboush et al 1993;Bogdanov et al 2008;Castro et al 2001;Ciulu et al 2011;Haydak et al 1942;White 1975).…”

“…In contrast, the following three research groups did not find any ascorbic acid in any honey sample. First, Castro et al (2001) added metaphosphoric acid in the mobile phase (a MeOH/H 2 O solution, 15/85 v/v) while using a C18-ODS column. The revelation was accomplished at 254 nm.…”