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HONEY BIOACTIVITY

Bioactivity Certification

We received our Honey Bioactivity certificate from the University of Texas at San Antonio's HONEY Pathway project. This certificate verifies the Antibacterial, Antioxidant and Biological Activity Levels found in our honey.

THE RESULTS - Sample ID: 23H-116

Biological Activity Level (BAL) Numbers of our Honey

19.5 mm - Antibacterial Activity (Zone of Inhibition (ZOI)
77% Antioxidant Activity (DPPH Radical Scavenging)
4.91% pH Level
16.5% Moisture Content
Amber - Honey Color (86-114 Pfund)
77% Biological Activity Level (BAL)

What do these numbers mean?

Honey has been used as a medicine by every civilization on earth due to its medicinal properties and biological activities originating from its nectar source. Honeybees forage on flower blossoms, tree secretions, and insect excretions to collect the nectar and convert it to honey using the enzymes within their abdomens. Since these nectars provide various chemical constituents such as phenolic acids, flavonoids, tannins, pigments, minerals, and vitamins, every honey’s biological activity differs from one another based on the abundance and ratios of these compounds. These components vary in accordance with the terroir, climate, foraging window, and honeybee strain. Therefore, we can conclude that every honey is created differently.

Dr. Ozturk’s research explores the medicinal potential of Texan honey varieties, introducing a comprehensive Biological Activity Level (BAL) formula that amalgamates these properties. The study presents a novel approach that integrates both physical and chemical properties, along with antibacterial and antioxidant potential, to gauge honey’s biological activity.

An array of honey samples from diverse regions across Texas underwent various methodologies to compute the BAL for each sample. Based on their BAL values, the honey samples were classified as having notably high or low bioactivity levels. Notably, honey from the San Antonio-Hill Country region exhibited particularly high BAL values. These findings underscore the remarkable medical-grade potential inherent in locally sourced Texas honey.

The honey analysis certificates contain the following measurements:

1.0 Physicochemical Properties

1.1. Color Detection
The color of honey is strongly related to the botanical origin of the honey sample and can be used to assess the quality of the honey. According to the United States Department of Agriculture, there are 7 colors of honey and honey color standard designations are expressed using the Pfund scale.

COLOR DESIGNATIONS OF HONEY
USDA COLORSTANDARD DESIGNATION COLOR RANGE PFUND SCALE (MM) SAMPLE RESULT RANGE
Water White <8 0 - 0.094
Extra White > 8 and < 17 0.094 - 0.189
White > 17 and < 34 0.189 - 0.378
Extra Light Amber > 34 and < 50 0.378 - 0.595
Light Amber > 50 and < 85 0.595 - 1.389
Amber > 85 and < 114 1.389 - 3.008
Dark Amber > 114 > 3.008

1.2 pH
The pH range of honey to be classified as acidic is between 3.2 and 4.5. The acidic nature of honey contributes to antimicrobial activity because it is low enough to inhibit bacterial pathogens. While most honey creates an environment to inhibit bacterial growth, not all the Table 1 - Color Designations of Honey USDA Color Standard Designation Color Range Pfund Scale (mm) Sample Result Range Water White ≤8 0-0.094 Extra White >8 and ≤17 0.094- 0.189 White >17 and ≤34 0.189-0.378 Extra Light Amber >34 and ≤50 0.378-0.595 Light Amber >50 and ≤85 0.595-1.389 Amber >85 and ≤114 1.389-3.008 Dark Amber >114 >3.008

Texas honey samples analyzed were acidic. The acidity value of the samples ranged from 3.73 - 6.76, with an average of 4.94. The low pH can inhibit the growth of bacteria and is an indicator of the overall quality of honey. The pH of honey sample was evaluated using the Accumet pH meter (Fisherbrand). 40% of honey solutions were prepared using 2g of honey sample diluted in 5mL DI water in borosilicate glass tubes.

1.3 Moisture Content
Honey is characterized by low moisture content and as a result, few bacteria can survive in this environment, therefore low water activity the greater antibacterial environment. The moisture content of the honey samples ranged between 14.40 and 22.00, with an average of 17.53% the honey samples are in range for what an acceptable moisture content should be which is typically around 17% - 18%. The amount of moisture affects various characteristics including the color, viscosity, flavor, density and more importantly, fermentation which affects the color of honey, changes its flavor, and alters its shelf life.

2.0 Antimicrobial Properties

2.1 Zone of Inhibition
The antibacterial activity of the honey samples was determined using the agar well-diffusion method on Staphylococcus aureus bacteria to measure the zone of inhibition after 18-h incubation at 37oC with the presence of honey samples. The zones higher than 16mm were considered as high antimicrobial activity according to the Clinical and Laboratory Standard Institute (CLSI) guidelines.

3.0 Antioxidant Properties

3.1 DPPH Radical Scavenging Assay (RSA)
The DPPH radical-scavenging assay (RSA) is based on the ability of antioxidants to block the free radical of DPPH (2,2-difenil-1-picril-hidrazil) and is used to measure the antioxidant activity of the honey samples. Due to its simplicity, DPPH assay has been the most used method to measure honey’s antioxidant activity. The RSA of honey sample was determined as a percentage based on the discoloration of DPPH after 30-minute incubation. The higher percentage RSA, the higher antioxidant activity of the honey sample.

4.0 Biological Activity Level (BAL)

PERCENTAGE BAL VALUE
> 81% Very High
61% TO 80% High
41% TO 60% Moderate
21% TO 40% Low
< 20% Very Low

BAL values of the TX honeys were calculated using a special formula including the three major properties of each honey sample and converted to percentages according to the highest BAL value among 193 samples. Honeys were classified using the percentage table.

HONEYBEES ARE DILIGENT CHEMISTS

Honey is produced through enzymatic processing of the nectar or honeydew honeybees collect from various plants. The invertase enzyme within the bees’ abdomens catalyzes the conversion of sucrose – table sugar within the nectar – into glucose and fructose. Honey is supersaturated in sugars, packed with beneficial chemicals, and, thus, possesses high nutritive value. It has been used both as a nutritious food and a remedy for various ailments throughout history. Wound healing, and treatment of gastrointestinal diseases, conjunctivitis, acute fever, and pain, are just some of the disorders honey can treat.

Honey is not only a supersaturated sugar solution, but also equipped with more than 200 various chemical and biological components.

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