CHAPTER - 5
Discussion
________________________________________________________________________
_____
The chapter deals with discussion of the results.
The thesis
ends with this chapter. The chapter interpretates and discusses the
results of the investigation on the physical properties of mineral and
biological waxes, and animal fat. The chapter starts with brief
introduction on waxes. The results of the investigations have been
discussed, correlating with biological and medical and technological
aspects. At the end of this chapter, the important conclusions drawn
from the investigations are presented.
__________________________________________________________________
_____
The different waxes such as Mineral wax (Microcrystalline wax
and Paraffin wax); biological wax (Beeswax); and animal Fat are taken
for study of physical properties using the techniques like FTIR, NMR,
GC-MS, XRD, microwave bench, network analyzer and impedance
analyzer. The results obtained are tabulated for comparison and
discussion.
The case wise critical review of results from various methods
revealed the behavior of different waxes and fat at wide range of
frequencies, which is useful in medical discipline and also in
technology electronics. From the present investigation, it is noticed
that the density of the samples is considerably less when compared
with that of water. Further, among the wax samples, the density of
paraffin wax is less, while the density of Beeswax and animal fat is
high. The heat treatment increases the density, irrespective of type of
wax (Table 5.1).
The visible region of electromagnetic radiation extends from 0.38
to 0.78 µm. The infrared region extends from the end of the visible
region at 0.78 µm to the microwave region with a wavelength of
~1mm. The infrared region is usually divided into three sections.

The section used mostly by material scientist is the mid infrared
region extending from 2.5 µm or 4000 cm-1 to
~ 50 µm
or
200 cm-1. The region between the visible and mid infrared is
called near infrared region. This region of the infrared has been
used for many applications, especially for quantitative analysis.
The region beyond ~50 µm (200 cm-1) is called the far infrared
region. This region is used for studying the low frequency
vibrations and some molecular rotations.
Common infrared spectrophotometers cover wave numbers raging
from 4000 to 200 cm-1. The spectrum is a plot of sample absorbance
or percent transmittance versus wave number. A grating or several
prisms
are
used
for
changing
wavelengths
and
for
monochromatisation. Sodium Chloride prisms are used for the 4000
to 600 cm-1 range, while cesium bromide prisms are used for 600 to
200 cm-1 range. However, the wavelength scale usually changes with
use, and therefore, frequent calibration is necessary.
Biological samples can be studied as mulls in Nujol. The mull is
prepared by grinding the sample to a fine powder and mixing with a
mulling agent to maker a paste. A thin layer of the paste is studied
between Nacl and other plates. Another method is the KBr disc
method in which the powdered sample is mixed with KBr and is
pressed into a clear disk which is mounted and examined directly.
Care must be taken while analyzing infrared spectra, because of
spurious bands present in IR spectra. A list of such bands have been
prepared, some of which are presented in Table 5.2. (Launer, 1962).
In the present investigation, the infrared spectra of powdered
samples of different waxes and animal fat were recorded at the room
temperature of 300C with a JASCO 3500 FTIR spectrophotometer.
In general, spectral analysis of tissues of biological systems
depends upon the material present and the analyze being sought. The
tissue itself is dominated by the spectrum of the macromolecular
components, which are present to the largest amount. If living tissue
is being examined, it is dominated by the water spectrum. For the
spectroscopic analysis, the biological material can be classified into
Four types:
1. Organic tissues such as muscle, brain, liver, kidney, heart, etc.
2. Mineralized tissues such as bone, calculi, gallstones, horn, hoof,
teeth and nail, etc.
3. Body fluids such as blood, urine, saliva, bile, spinal fluid, etc.
4. Waxes (mineral and biological) and fats
The main aim of the IR spectroscopic analysis of the tissues is the
determination of structure and combinations of organic and inorganic
constituents of the tissues. Further, it is an attempt to investigate
what variation has occurred in the spectrum of normal tissue. What is
the effect of some physiological change, and what change might occur
in the normal composition with ageing? It might be of interesting to
analyze for a particular component which changes in response to some
physiological stress. Finally, infrared spectra of tissues may be
utilized to determine whether there are changes in the structure of
various components as part of the study of the mechanism of the
reactions within the living cell.
The present investigation is aimed to analyze the principal
constituents of some waxes in normal and heat treated conditions,
and fat in normal condition.
For the convenience of analysis the IR spectrum can be divided
into three regions.

The first region 4000 to 3000 cm-1 concerned with water and
hydroxyl group. This region is of considerable interest to
present study because it reveals the nature of hydrogen
bonding.

The second region is 3000 to 1500 cm-1, wherein bands for
functional groups are seen. In this region, major IR absorptions
of waxes and fat occur.

The third region (1500 – 200 cm-1) has significant importance in
the context of biological apatite and other salts. The spectra of
waxes and fat indicate the presence of bands characteristics of
some functional groups concerned with the waxes and fat.
These are the major bands which can be attributed to the
organic
component of the waxes and fat.
In the case of waxes and fat of the present investigation, FTIR
Spectroscopy confirms that some functional groups are absent after
heat treatment. Even after heat treatment, the main compounds are
present. The presence of peaks at 2920 cm-1, 1460+10 cm-1, 1375 +10
cm-1, 889+10 cm-1, 717+10
cm-1 is characteristic of wax. The
additional peaks such as 3605 +10 cm-1, 3390+10 cm-1, 2955+10 cm-1
and 1736+10cm-1 suggest the sample is a beeswax. Similarly, the
presence of 1163+5cm-1 peak which refers to triglycerides tells that
the sample is a fat (Table 5.3).
Beeswax, microcrystalline wax, paraffin wax and animal fat
contain large number of compound as evident from the data obtained
from gas chromatography and mass spectrum (GC-MS). The provision
is made for the temperature to go up to 300
0C.
The pressure is
maintained low such that the compounds having higher boiling point
can easily eluded. The data of GC-MS of beeswax signifies that it
contains alkenes, acid and alkane groups. Similarly the data of GCMS of microcrystalline wax and paraffin wax shows that it contains
only alkanes. The GC-MS report of animal fat tells that it contains
acids and alkanes (Table 5.4).
The spectra of 1H1 (proton) and
13C
NMR justify the same by
showing the peaks at aliphatic region for all waxes and fat. For
beeswax and fat, the peaks are seen at 4 ppm in1H1 and in between
70-80 ppm for
1H1
13C
NMR, indicating the presence of acid group. The
NMR of microcrystalline wax and paraffin wax indicates the
presence of triplet at the end, which specifies the end molecule
contains 3 hydrogen atoms.
X-ray diffraction of waxes signifies that they are crystalline in
nature (Figures. 4.5.1 to 4.5.7). The crystalline nature of wax is also
proved from the GC-MS data. The physical properties of eluded
compounds are gathered, which reveals the crystalline nature of
waxes under study (Table 5.4). Further, it is found that more number
of the compounds are crystalline in nature. The components of animal
fat, which are predicted by GC-MS, reveal that only few compounds
are crystalline in nature. From x-ray diffractograms (XRD) of waxes of
the
present
investigation,
it
is
evident
that
mineral
waxes
(microcrystalline wax and paraffin wax) exhibit sharp peaks, when
compared to beeswax and animal fat. The sharp peaks indicate that
more number of compounds are crystalline in nature. The peaks
present in beeswax are found to be little bit wider than the mineral
wax. It shows some compounds of the beeswax are crystalline in
nature. The above Table reveals the information on the percentage of
crystalline compound, out of eluded compound. In conclusion, one
can say that the wax, whether it can be mineral wax or biological wax,
contains a large number of amorphous and crystalline compounds.
Therefore it is not worth to draw conclusion that the wax or fat is only
crystalline or amorphous material.
The XRD of heat treated waxes and animal fat shows a decrease
in
the inter planer distance, which indicates the shifting of the
molecules. As the inter planer distance decreases the volume of the
molecule is also decreases, which reveals that the density decreases
after heat treatment.
The V-I characteristic curves are linear and pass through the
origin (Figures 4.6.1 to 4.6.7.). As the curves are linear, they are
ohmic in nature. It is noticed from the present study on electrical
properties that after heat treatment the resistance decreases. In other
words, one can say that the conductivity increases. It is in accordance
with the general theory of resistivity that the resistivity decreases as
density increases. The waxes follow the same rule. After heat
treatment, the increase of density shows the decrease in resistivity.
The resistance of microcrystalline wax is found to be more, which
indicates it is a good insulator compared to the other samples.
The dielectric data of different samples that obtained by using
the
microwave bench at 8.9 GHz shows a decrease in dielectric
constant of beeswax and paraffin wax after heat treatment, where as
the dielectric constant is increasing for microcrystalline wax after heat
treatment.
It is interesting to note that the values of dielectric constant at
the frequency of 8.9 GHz that are measured using microwave bench
and sophisticated network analyzer are in good agreement. This
reveals the fact that in the absences of sophisticated expensive
instrumentation (network analyzer), an inexpensive microwave bench
solves the purpose.
The study of dielectric constant of different samples at
microwave range frequencies in x-band and p-band regions reveals that
dielectric constant is decreasing with the increase of frequency in all
cases (Tables 5.9).
The
dielectric
data
of
different
samples,
studied
using
impedance analyzer, reveals a decrease in impedance and dielectric
constant with the increase of frequency (Table 5.10).
The dielectric study on waxes and fat in wide range of
frequencies staring from 10 kHz to 18 GHz presents some following
interesting features:

There is no significant dielectric decrement in the
frequency range of 10 kHz to 1MHz. Further, it seems
that electrical polarization mechanisms are not so
pronounced.


Also same is the observation in x- band frequency region.
But a significant decrease in dielectric constant of wax
and fat samples is noted as frequency in p-band (12.4
GHz -18 GHz) region increases.

In general, it is obvious from the dielectric spectra of
waxes and animal fat under study that the value of
dielectric constant is low, varying from about 8 to 3 in the
wide frequency range of 10 kHz to 18 GHz.
Table 5.1 Density of different wax samples using single pan
balance
S.No
Sample name
Condition of
sample
Density, ρ
(gm/cm 2)
1
Beeswax
normal
0.8922
2
Beeswax
heat & cool
0.9334
3
Microcrystalline wax
normal
0.8706
4
Microcrystalline wax
heat & cool
0.8983
5
Paraffin wax
normal
0.7700
6
Paraffin wax
heat & cool
0.8083
7
Fat
normal
0.8974
Table 5.2 some common spurious absorption bands in infrared
spectra
S.NO Wave number Wave Length
Compound
Source
-1
(Cm )
(μm)
or group
1
2
3700
3650
2.70
2.74
H2 O
H2
Any Source
Any Source
Hydrogen bonding in
water, usually in KBr
discs.
3
3450
2.90
H2 O
4
2350
4.26
CO2
Atmospheric absorption
5
2000-1430
5–7
H2 O
Atmosphere
6
1640
6.10
H2
Water of crystallization
7
1430
7.00
CO3
Contaminant in halide
window.
8
1360
7.38
NO3
Contaminant in halide
window.
9
1270
7.90
SiCH3
Silicon oil or grease
10
1110-1000
9 – 10
SiO
Glass
11
667
14.98
CO2
Atmosphere
Table 5.3 Comparison of FT IR data of wax samples
S.NO
Sample name
Condition of
sample
1
Beeswax
normal
2
Beeswax
heat & cool
3
Microcrystalline
wax
normal
4
Microcrystalline
wax
5
6
7
Paraffin wax
Paraffin wax
Fat
heat & cool
normal
heat & cool
normal
Range of characteristic
wave numbers in cm-1
3605, 3449, 3390,
2966, 2646, 2334,
2150, 1896, 1732,
1454, 1377, 1180,
1122, 958, 920, 837,
729
3390,2956, 2851,
2636, 2357, 1738,
1464, 724
2920, 2733, 2334,
1633, 1462, 1377,
1180, 889, 717
2928, 2339, 1599,
1462, 1377, 1180,
889, 721
2918, 2336, 1898,
1462, 1379, 1128,
889, 727
2916, 2339, 1898,
1469, 1379, 889, 721
3466,2918, 2332,
1730, 1468, 1161,
968, 891, 721
Table 5.4 Comparison of different samples with reference to
compounds present through GC-MS
S.No.
1
2
3
4
Sample
name
Beeswax
Microcrystall
ine wax
Paraffin wax
Fat
Condition
of sample
normal
normal
normal
normal
Name of the compounds present within the
sample
Cyclohexadecane, palmitic acid, octadecane,
Heneicosane, octadecenoic acid, stearic acid,
Docosane,
Tricosane,
Tetracosane,
Pentacosane,
Hexacosane,
Heptacosane,
Octacosane, Nonacosne, Triacontane
Hexadecane,
Octadecane,
Nanodecane,
Eicosane, Heneicosane, Docosane, Tricosane,
Tetracosane,
Pentacosane,
Hexacosane,
Heptacosane,
Nonacosne,
Triacontane,
Hentriacontane, Dotriacontane, Tritriacontane.
Heneicosane,
Docosane,
Tetracosane,
Pentacosane,
Octacosane
Nonacosne,
Dotriacontane,
decyltetracosane.
Tricosane,
Hexacosane,
Triacontane,
Tritriacontane,
n-hexane, acetic acid, Tetradecanoic acid,
palmitoleic acid, palmitic acid, Margaric acid,
Oieic acid, Stearic acid, Tetratetracontane.
Table 5.5 Data on crystalline compound in wax and fat samples
% of crystalline
compounds
present within
the sample
Name of the crystalline compounds
present within the sample
S.NO
Sample name
1
Beeswax(nor)
2
Microcrystalline
wax (nor)
Cyclohexadecane, Heneicosane
Docosane, Tricosane, Tetracosane,
Hexacosane, Heptacosane,
Nanodecane, Eicosane, Heneicosane,
Docosane, Tricosane, Tetracosane,
Hexacosane, Heptacosane,
Hentriacontane
3
Paraffin wax
(nor)
Heneicosane, Docosane, Tricosane,
Tetracosane, Hexacosane,
Dotriacontane, decyltetracosane
4
Fat (nor)
Margaric acid
46
56
58
11
Table 5.6 A Comparison on resistivity of different samples.
S.NO
Sample name
Condition of
sample
Resistivity
ρ(x107Ω.m)
1
Beeswax
normal
906 - 956
2
Beeswax
heat & cool
315-358
3
Microcrystalline wax
normal
1216 - 1296
4
Microcrystalline wax
heat & cool
219 - 235
5
Paraffin wax
normal
192 - 231
6
Paraffin wax
heat & cool
39 - 45
7
Fat
normal
452-510
Table 5.7 Data on Dielectric constant of different wax samples
obtained from Microwave bench at 8.9 GHz.
S.NO
Sample name
Condition of
sample
Dielectric
constant
(ε')
1
Beeswax
normal
6.19
2
Beeswax
heat & cool
6.09
3
Microcrystalline wax
normal
5.84
4
Microcrystalline wax
heat & cool
6.01
5
Paraffin wax
normal
5.30
6
Paraffin wax
heat & cool
4.43
7
Fat
normal
6.55
Table5.8 Comparison of dielectric constant of different samples
using microwave bench and network analyzer at 8.9 GHz.
Dielectric
constant from
microwave bench
(ε')
Dielectric constant
from Network
analyzer
(ε')
S.NO
Sample name
Condition of
sample
1
Beeswax
normal
6.19
6.18
2
heat & cool
6.09
6.11
normal
5.84
5.88
heat & cool
6.01
6.05
5
6
Beeswax
Microcrystalline
wax
Microcrystalline
wax
Paraffin wax
Paraffin wax
normal
heat & cool
5.30
4.43
5.27
4.45
7
Fat
normal
6.55
6.57
3
4
Table 5.9 Percentage decrement of dielectric constant
different samples at frequency range 8.2 GHz – 18 GHz.
Dielectric constant
S.No
Sample
At
At
8.2GHz
18.0GHz
of
Percentage
Difference
decrease
1.
Beeswax (nor)
6.6
2.65
3.95
59.8
2.
Beeswax (h&c)
6.26
2.63
3.63
57.9
3.
Microcrystalline wax (nor)
6.35
2.57
3.78
59.5
4.
Microcrystalline wax (h&c)
6.8
2.55
4.25
62.5
5.
Paraffin wax (nor)
5.59
2.02
3.57
63.8
6.
Paraffin wax (h&c)
4.89
2.01
2.88
58.9
7.
Fat (nor)
6.77
2.83
3.94
58.2
Table 5.10 Variation of dielectric parameters with increase of frequency
range(103-105) Hz computed through Impedance Analyzer.
S.NO
Sample name
Condition
Variation of
Variation of
Impedance, Z
Capacitance, Cp
(x10 6Ω)
(x10-12F)
Variation of
Dielectric
Constant
ε'
1
Beeswax
normal
9.73-0.06
3.56-3.17
7.64-6.81
2
Beeswax
heat & cool
10.7-0.06
3.32-3.05
7.14-6.55
3
Microcrystalline wax
normal
17.16-0.08
2.65-2.45
7.49-6.91
4
Microcrystalline wax
heat & cool
13.14-0.07
3.75-3.45
7.20-6.63
5
Paraffin wax
normal
19.03-0.1
1.82-1.55
6.97-5.96
6
Paraffin wax
heat & cool
16.62-0.08
1.48-1.32
5.67-5.06
7
Fat
normal
4.47-0.1
2.45-2.07
9.69-7.72
Table 5.11 Percentage decrement of dielectric constant of
different samples at frequency range 10kHz – 1MHz.
S.No
Sample
Dielectric constant
At 10kHz
At 1MHz
Difference
7.38
6.81
0.57
Percentage
decrease
1.
Beeswax (nor)
2.
Beeswax (h&c)
6.91
6.55
0.36
5.21
3.
Microcrystalline wax (nor)
7.19
6.91
0.28
3.89
4.
Microcrystalline wax (h&c)
7.02
6.63
0.39
5.56
5.
Paraffin wax (nor)
6.42
5.96
0.46
7.17
6.
Paraffin wax (h&c)
5.48
5.06
0.42
7.66
7.
Fat (nor)
8.18
7.72
0.46
5.62
7.72
Figure 5.1 density of different types of waxes and fat
8.2 GHz
Figure 5.2 variation of dielectric constant of different types of waxes
and
fat at 8.2 GHz using network analyzer.
Figure 5.3 variation of dielectric constant of different types of waxes
and
fat at 18 GHz using network analyzer.
Figure 5.4 comparision of dielectric constant of different types of
waxes
and fat at 8.9 GHz using microwave bench and network
analyzer.
Figure 5.5 variation of dielectric constant of different types of waxes
and
fat at 10 kHz using impedance analyzer.
Figure 5.5 variation of dielectric constant of different types of waxes
and
fat at 10 kHz using impedance analyzer.
Conclusion
From the present study the following conclusions are drawn.

The density or specific gravity of waxes and of animal fat is found
to be less than that of water. After heat treatment, the density
increases in all types of waxes of the investigation.

From FTIR study, it is concluded that all waxes and animal fat are
heterogeneous in nature. Beeswax and animal fat contains
alkanes, alkenes and acid group, while in mineral wax (paraffin
and microcrystalline wax) alkanes and alkenes groups are present.

The above statement is also confirmed by 1H1 and
13C
NMR study.

Also GC-MS technique strongly confirms the result obtained from
spectra of FTIR and NMR studies.

XRD study suggests that beeswax of the present investigation are
crystalline nature. However, the typical crystal structure combines
crystalline and amorphous characteristics.

The wax, whether it is mineral wax or biological wax, contains a
large number of amorphous and crystalline compounds.

It is noticed from the present study on electrical properties that
after heat treatment the resistance decreases.

The resistance of microcrystalline wax is found to be more, which
indicates it is a good insulator compared to the other wax samples.

There is no significant dielectric decrement in the frequency range
of 10 kHz to 1MHz. Further, it seems that electrical polarization
mechanisms are not so pronounced. Also same is the observation
in x- band frequency region.

But a significant decrease in dielectric constant of wax and fat
samples is noted as frequency in p-band (12.4 GHz -18 GHz)
region increases.

In general, it is obvious from the dielectric spectra of waxes and
animal fat under study that the value of dielectric constant is low,
varying from about 8 to 3 in the wide frequency range of 10 kHz to
18 GHz.