Sample Pfund Scale Grade LH1 LH2 LH3 LH4: Figure. Pfund Scale For Honey (Source: WWW - Oxfordhoney.uk)

2.
1 Materials and Methods

Four honey samples are prepared. The samples are of multi-floral origin with tallow
and willow being among the major sources of pollen. The honey color is designated by
measuring the optical density and expressing it in terms of millimeters (Pfund scale). The
corresponding grade is then read from the manual provided with the instrument and the honey
samples are labeled based on the Pfund value of each. For example, honey sample with
Pfund scale 80, 65, 79 and 77 mm are labeled LH1, LH2, LH3 and LH4 respectively.
Figure. Pfund Scale for Honey

(source: www.oxfordhoney.uk)
Table. USDA color designation
Sample Pfund scale

LH1
LH2
LH3
LH4
Grade
80
65
79
77
Light amber
Light amber
Light amber
Light amber
2.2.2 Spray Drying of Liquid Honey Using Retrograded Starch as Drying

Agent
A solution comprising 20% honey, 30% retrograded corn starch and 50% water (by
weight) is prepared (at room temperature). The proportions are determined based on
preliminary studies. The solution is prepared by mixing continuously using a magnetic
1
stirrer until the honey dissolved completely in the solution (10 mins). The mixture is then
sonicated using a laboratory scale ultrasonic for 5 minutes at 80% amplitude (pulse on 2 and
pulse off 1). The solution is placed in an ice bath to prevent any temperature rise in the
solution. Sonication is done to ensure that the particles are smaller in size and well
dispersed in the solution. The honey solution is spray dried into dry powder using spray
dryer. The honey solution is pumped at a flow rate of 9 mL/min and spray dried at 200
C. The spray dried honey powder is stored at 4 C until analyzed.
2.2.2. Characterization of Spray-dried Honey Powder
The spray-dried honey powders are labeled based on the honey they are spray-dried from
Thus honey powder produced is labeled HP1, HP2, HP3 and HP4 to signify its production
from LH1, LH2, LH3 and LH4 honey samples respectively. Then, for the
characterization, we measure the color of honey powder, moisture content using moisture
analyzer, HPLC sugar analysis and SEM.
2.2.2.1 Color
The color of the honey powder is measured. The measurements are made in
triplicate and the means and standard deviations are reported.
2.2.2.2 Moisture Content
The moisture of the honey powders is measured using a moisture analyzer and reported
as the means of the triplicate measurements along with the standard deviations.
2.2.2.3 HPLC Analysis for Sugar Content
The sugar content of the honey powders is determined using an HPLC technique.
Here below the method :
Half percent solution of the honey powder samples is prepared by dissolving

0.5 g of honey powder in 100g of distilled water.
Of this solution, 15 L is injected into HPLC column (Waters). For

analysis of the glucose and fructose, the CarboPac PA10 column (4 x 250
mm) is used with 112 mM NaOH as the mobile phase at ambient room
temperature. They are detected using an EC detector system.
The contents of glucose, fructose, sucrose and maltose in liquid honey are calculated
based on the ratio of integrated peak areas using commercial glucose, fructose, sucrose and
maltose as the standard compounds. Triplicate analysis is conducted for each sample and
2
results reported as the mean along with the standard deviation.
Figure. HPLC instrument

(source: oxford-instruments.com)
2.2.2.4 SEM Analysis for Particle Morphology

The Scanning Electron Microscope (SEM) uses a focused beam of high-energy
electrons to generate a variety of signals at the surface of solid specimens. The signals that
derive from electron-sample interactions reveal information about the sample including
external morphology (texture), chemical composition, and crystalline structure and orientation
of materials making up the sample. The morphology of honey powders is determined using
Scanning Electron Microscopy (SEM) using an acceleration voltage of 5 kV.
Figure. SEM Instrument

(source: serc.carleton.edu)

Sample Pfund Scale Grade LH1 LH2 LH3 LH4: Figure. Pfund Scale For Honey (Source: WWW - Oxfordhoney.uk)

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2.

1 Materials and Methods

Figure. Pfund Scale for Honey

Sample Pfund scale

2.2.2 Spray Drying of Liquid Honey Using Retrograded Starch as Drying

Half percent solution of the honey powder samples is prepared by dissolving

Of this solution, 15 L is injected into HPLC column (Waters). For

results reported as the mean along with the standard deviation.

Figure. HPLC instrument

2.2.2.4 SEM Analysis for Particle Morphology

Figure. SEM Instrument

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