Honey Analysis

ANALYSIS

Fall Sample:

Your sample would be classified as a MIXED FLORAL HONEY because none of the pollen types reach the required minimum level of 45%. When you look at the pollen counts in Table 1, you can see that it appears that the bees visited a wide variety of flowers. However, it appears that a few of the major nectar sources used in your honey are from a plant in the lily family (it appears to be a species of Yucca), buttonbush, crepe myrtle, clover, and Chinese tallow. There are a number of other minor types of pollen represented and by inference those flowers may have contributed minor amounts of nectar to form your honey. There are also two windborne pollen types (ash and elm), which may have blown into the hive and become part of the honey. Overall, this appears to be a typical mixed floral honey from areas of Central and North Texas. The pollen concentration value of 82,591 pollen grains per 10 grams of honey is within the expected normal range.

EXTRACTION PROCEDURE:

To conduct a pollen study of raw honey we first must dilute it before the pollen can be removed for analysis. For our study, we use a 10g sample of raw honey for the analysis. The sample of raw honey is diluted with 10 ml of distilled water and 150 ml of ETOH, and then heated to 100o F to ensure a complete mixture. This is a technique that we developed and has now been adopted by most others (Jones and Bryant, 2004, The use of ETOH for the dilution of honey Grana 43: 174–182).

Next, we add one tablet containing a total of 18,583 Lycopodium spores to enable us to conduct a pollen concentration study for each sample. We use these lycopod spores because they are not utilized by bees for any purpose and thus we do not have to worry about these being found in natural honey sources. Once these initial stages are complete, the pollen sample is dehydrated with glacial acetic acid and then heated in a mixture of a sulfuric acid and acetic anhydride. This chemical treatment, called acetolysis, is designed to remove lipids, waxes, and cytoplasm thereby making the pollen easier to identify. 

Once the acetolysis process is complete, each sample is again dehydrated in glacial acetic acid and treated with a series of distilled water rinses. The resulting pollen residue is stained to create contrast for microscopic analysis and photography. Finally, we mix a few drops of glycerin into the sample and mount one drop of it on each microscope slide for analysis. To ensure an accurate representation of the overall sample we stir the sample for one minute on a Vortex stirrer before removing each drop for analysis. Our laboratory experiments and published results have demonstrated that this technique ensures that each drop is a true reflection of the original sample.

Analysis of a honey sample follows a two-step procedure. First, the sample is scanned at 400x under a microscope, initial identifications are made of each pollen type, and key photographic images are taken of each pollen type. During this procedure if a pollen grain is not one we are familiar with, we will compare it with our extensive modern pollen reference samples on file in our laboratory in hopes of finding a match. Second, a quantitative pollen count is conducted for each sample to determine the pollen types present and the frequency of each taxon. 

A statistically valid quantitative pollen count of 200+ pollen grains is conducted for each sample as originally recommended for honey specimens in 1978, by Louveaux, Maurizio, & Vorwohl (Bee World, Vol. 59:139-157). Quantitative counts are used because testing has shown that these offer an accuracy of greater than 95% as to the actual composition of pollen taxa within a given honey sample. 

We have followed the reporting system recommended by Louveaux et al. (op. cit.) and others who stress that pollen results should be listed according to percentage classes rather than actual percentages when counts of between 200-1200 grains per sample are conducted. We show the actual percentage counts for general reference but these are not deemed totally accurate for honey samples until a total count in excess of 1,200 pollen grains per sample is reached. We rarely count this many pollen grains for a honey sample because in most cases it is not needed and because larger counts add cost and time considerations.