Detection of Foreign Enzyme Addition into the Adulterated Honey

Abstract

Natural honey contains several enzymes, which are produced by bees (salivary secretion) and some are found in the nectar or pollen. The most important enzymes are amylases, invertases, glusidases, catalases, fosfatases and other. The activity of diastase (α-, β-, γ-amylase) is the important quality parameter of honey, according to the Directive 2001/110/CE the diastase activity (diastase number) must not be less than or equal to 8, for some kinds of honey also higher or equal to 3 (in these cases the HMF must not be higher than 15 mg/kg). Diastase is used as a marker to evaluate the freshness or the heat damage of honey. When honey is adulterated by addition of inverted sucrose or hydrolysed starch namely high fructose corn syrup (HFCS), then such dilution of honey leads to the reduction of diastase number. Such adulteration can be masked by addition of foreign amylases, e.g. bakery mould amylases. Recently several suspect samples of honey with inconsistent diastase number were found in the market. The possibilities of detection of foreign amylase addition based on the comparison of diastase determination using the Schade and Phadebas procedures are evaluated. The both tests are based on the determination of hydrolytic activity (the Schade number is expressed as g of starch hydrolysed 1 h at 40°C per 100 g honey), but the results depend on the substrate used for the trial (according to the standard procedure an insoluble blue dyed cross-linked type of starch should be used). The results of Schade test are therefore often affected by the choice of substrate. The model samples of honeys with addition of foreign amylase (Aspergillus oryzae) were analysed, the methods of adulteration detection based on the substrate specificity of enzymes is proposed.

The Unique Protein Composition of Honey Revealed by Comprehensive Proteomic Analysis: Allergens, Venom-like Proteins, Antibacterial Properties, Royal Jelly Proteins, Serine Proteases, and Their Inhibitors

“…Amylase is similar to HBG3 produced from the hypopharyngeal glands . Despite uncertainty, amylase activity in honey has become a parameter used to evaluate the freshness of honey, and importantly, amylase can potentially be added by producers to mask syrup adulteration of honey . In this study, we analyzed only authentic honeys and identified honeybee α-amylase as two isoforms with 99% identity; one less abundant isoform was not detected in three honeys.…”

“…19 Despite uncertainty, amylase activity in honey has become a parameter used to evaluate the freshness of honey, 75 and importantly, amylase can potentially be added by producers to mask syrup adulteration of honey. 76 In this study, we analyzed only authentic honeys and identified honeybee α-amylase as two isoforms with 99% identity; one less abundant isoform was not detected in three honeys. In addition, the protein hit of amylase that was detected in all the samples analyzed showed very low proteomic variation; again, however, the varying total protein contents (Table 2) in honeys should produce different amylase abundance levels or enzyme activities.…”

The Comparison of Physicochemical Parameters, Antioxidant Activity and Proteins for the Raw Local Polish Honeys and Imported Honey Blends

“…The amylase in honey is considered to be mainly of bee origin and is secreted by the salivary and hypopharyngeal glands [ 31 ]. However, the presence of proteins with amylolytic activity derived from plants or microorganisms cannot be ruled out [ 32 ]. Gels stained for acid phosphatase activity are shown in Figure 1 c. In all honey samples, the band corresponding to acid phosphatase was detected in the same place at the bottom of the gel.…”

The Effect of Honey Harvesting Age in the Area of Calliandra Plant to Diastase Enzyme Activity, Hidroximetilfurfural (HMF) and Acidity

“…Faktor lain yang menyebabkan nilai enzim diastase adalah perbedaan pH alami pada madu dan pola pencarian nektar (Voldřich et al, 2009).…”