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Safety Assessment of Shellac as Food Additive through Long Term Toxicity

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Trends in Biosciences 10(2), Print : ISSN 0974-8431, 733-740, 2017
Safety Assessment of Shellac as Food Additive through Long Term Toxicity Study
SANJAY SRIVASTAVA & NANDKISHORE THOMBARE*
Processing and Product Development Division, ICAR-Indian Institute of Natural Resins and Gums,
Namkum, Ranchi
*email : nandkishore.icar@gmail.com
ABSTRACT
Shellac is the natural resin of insect origin cultivated on
specific host plants. In view of non-availability of sufficient
toxicological data on shellac, the chronic toxicity study
was conductedin which shellac was administered orally by
mixing in powdered feed at 5000, 10000 and 20000 ppm to
Wistar rats for a period of 180 days.The investigations on
clinical signs/ behavior, feed consumption, body weight
gained, organ body weight ratio of animals, necropsy and
histopathological tests were conducted and showed no
significant difference between control and treated animals
from all dose levels. But, significant impact of shellac was
observed at mid and high dose animal groups altering
various haematological parameters. As, no significant
variations in any of the parameters for male and female
group were observed at low dose, the administration of
shellac up to 5000 ppm in feed can be considered as safe
and without any toxic manifestation.
Key words Lac, resin, food additive, E904, Wistar rats
Shellac is derived from the hardened protective
secretion of the lac insect Kerria spp. feeding on trees and
bushes cultivated in India, China, Thailand and Indonesia
(Baboo and Goswami, 2010). In India, lac cultivation is done
mainly through culture of Indian lac insect Kerria lacca on
hosts such as kusum (Schleichera oleosa), palas (Butea
monosperma) and ber (Ziziphus mauritiana). Lac
encrustations on the twigs of host trees are removed by
scrapping, either manually or by machines. Lac resin, thus
obtained is known as sticklac. It contains number of non-
resinous impurities like wood particles, insect body,
wax,dye, etc., which are to be removed. The resin thus
obtained is called seedlac which is further refined and
converted into commercially acceptable form, known as
shellac.
Shellac consists of resin as the major component
along with wax and very small quantity of coloring
component (Bose et al., 1963; Wadia et al., 1969). The resin
is polyester (Figure 1) comprising a mixture of long chain
hydroxy and sesquiterpenic fatty acids. Resin can be
broadly resolved into two fractions on the basis of solubility
in ether: ether insoluble - hard resin (approx. 70%) and ether
soluble - soft resin (approx. 30%) (Sengupta, 1970).
Component acids present in the resin portion of shellac
are Aleuritic acid (30-35 %), Laccijalaric acid (22-26 %), epi-
Laksholic acid (12-15%), Jalaric acid (8-10 %), Butolic acid
(6-8 %), Shellolic acid (8-10 %), Myristic acid (traces) and
Palmitic acid (traces).Besides this, Wax (4-5%) and coloring
pigment, erythrolaccin (traces) are also present in shellac
as co-components (Rangaswami and Sen, 1952).
Shellac is the only natural resin of insect origin, mostly
produced in India, Thailand, China, Vietnam, etc. which is
exported to different parts of the world including European
countries and used in glazing and confectioneries with the
E number, E904. Being a natural product it finds widespread
applications in food(Phan et al., 2008; Chauhan et al., 2011;
Wan-Shin et al., 2014; Chitravathi et al., 2014; Tariq et al.,
2011; Valencia et al., 2009;USDA NOP, 2014) and
pharmaceuticals industries (Limmatvapirat et al., 2004;
Pearnchob et al., 2004; Stummer et al., 2010).
In view of the limited toxicological studies available
on the safety aspects of shellac, U.S. FDA and European
Union (EU) had permitted the use of shellac as a food
additive. The toxicological studies on shellac include acute
and short term oral toxicity tests, mutagenicity and
reproductive toxicity evaluation, etc. For acute oral toxicity
test (Levenstein, 1980)shellac was administered to the albino
rats with a single gavage dose of 5 g/kg body weight. No
deaths were recorded during the observation period of 14
days. In the short term toxicity studies of shellac, rats were
fed with diets containing 2% shellac for 90 days and
compared with a control (Buchloch, 1979). The investigator
reported that shellac had no effect on the appetite and
body weight gain of test animals and no residues of shellac
were detected in the test animal faeces. Rats from treated
and control group were studied for pathological and
histological examinations at the end of the study. Gross
examination revealed that no significant changes were
observed due to administration of test item (Buchloch,
1979). Reports are also available on mutagenicity and
reproductive toxicity of shellac (Okamoto, 1986;
Limmatvapirat et al., 2008) endorsing its non-toxic
manifestation, but no report is yet available on long term or
chronic toxicity of shellac. As per new safety requirement
of EU, European Food Safety Authority (EFSA) has pointed
out the need of detailed toxicological studies on shellac
and has called for long term toxicity studies data to keep it
in food additive list. Keeping in view the diversified
application of shellac but inadequate data on its toxicology,
the chronic toxicity study was undertaken to assess the
systematic toxicological effect of the shellac.
MATERIALS AND METHODS
Handmade shellac was obtained from M/s Gupta
Brothers, Ranchi, Jharkhand, India.
Analysis of the test sample
Major components,such as soft resin, hard resin and
wax content of the shellac sample to be used for the study
were analyzed by the standard methods (Rangaswami and
Sen, 1952). The test material was tested for physico-chemical
parameters through an ISO 9001:2008 certified quality
evaluation laboratory of the Indian Institute of Natural
Resins and Gums, Ranchi, as per IS 6921:1973 (Indian
standard for method of sampling and test for lac and lac
734 Trends in Biosciences 10 (2), 2017

Table 1. Physico-chemical parameters of shellac (test The animals were housed individually in standard
sample) polypropylene cages, with stainless steel top grill having
provision for pelleted food and drinking water in the bottle.
Sr.No. Parameters Tested Report Method of Paddy husk was used as bedding material and changed at
test least twice a week. Identification of animals was done by
1 Color Index 10 cage card and picric acid body marking. Certified rodent
feed, manufactured by Ashirwad India, Ltd., and reverse
2 Flow (mm) 80
osmosis water was provided ad libitum to all animals. The
3 Heat Polymerization 66 stability of test item in selected diet was determined and
IS
Time (min.) 6921:1973 found stable up to 8 days in diet when stored at room
4 Acid value 77 Indian temperature. Animals were maintained and housed at 22± 2
5 Moisture content (%) 1.88 Standard for, °C, with relative humidity 30-70% and day/night cycles of
Method of 12 hour light and 12 hour dark. Adequate fresh air supply
6 Impurity (%) 0.78 sampling
with regular air changes was maintained in the experimental
7 Ash (% by mass) 0.5 and test for
lac and lac room.
8 Matter insoluble in 1.25 products
hot alcohol (% by
Observations
mass) During the acclimatization period, all rats were
9 Lead (ppm) 1 observed at least once daily for any abnormalities. Only
rats that were suitable for use were assigned to this study.
10 Arsenic (ppm) 1
Females used in this study were nulliparous and non-
11 Softening point (oC) 65-70 pregnant. General examination of all the rats was carried
out prior to the test item administration on Day 1 and daily
products, Bureau of Indian Standard, New Delhi, India).
thereafter during the treatment period. Rats were observed
FT-IR spectrum wasobtained using Shimadzu Corpn., for changes in skin and fur, eyes, mucous membrane,
Japan, IR-Prestige 21, with optical system that gives data occurrence of secretions and excretions, changes in gait,
collection over a total range 7500–350 cm-1. posture and presence of any abnormal behavior. Similarly,
TOXICOLOGICAL STUDIES all rats were observed for morbidity and mortality once daily.
Individual body weights were recorded before the
Test system
administration of test item (day 1) and once weekly thereafter.
The chronic toxicity studies were conducted at CSIR- Fasting body weights were recorded prior to sacrifice.
Indian Institute of Toxicological Research (IITR), Lucknow, Fasted body weights were used for calculation of
to assess the systemic toxicological effects of the test item, organ: body weight ratios. The food consumption was
shellac, by administering orally in feed to Wistar rats as per measured at weekly intervals during in-life phase of the
OECD 452 (OECD guideline for the testing of chemicals no. experiment.
452, Chronic Toxicity Studies. Adopted on September 7,
Clinical pathology investigations
2009).Ethical Committee approval was obtained from
Institute Animal Ethics Committee, CSIR-IITR, which Blood collection
followed the guidelines of Committee for the Purpose of
Blood samples were collected at the end of the
Control and Supervision of Experiments on Animals
treatment period. All rats were fasted overnight (water
(CPCSEA), Ministry of Environment, Forest and Climate
allowed) before blood collection. The blood samples were
Change, Government of India. The doses of test item were
collected under anesthesia just before sacrifice for
decided on the basis of acute (Levenstein, 1980) and short-
necropsy. Blood was collected into K2 EDTA tubes for
term toxicological studies(Buchloch, 1979) carried out earlier.
haematology and tubes without anticoagulant for clinical
Accordingly the test item was administered for a period of
chemistry.
180 days, to three groups of Wistar rats at the levels of 5000
ppm, 10000 ppm and 20000 ppm by mixing in the powdered Hematology
feed. In addition, a concurrent control group that received Hematological parameters (mentioned in table 2 & 3)
only the powdered feed was also included. were determined using Sysmex XT 1800iv hematology
Test animal and husbandry analyzer.
Random bred, 6 week old male and female albino Clinical chemistry
Wistar rats (Rattus norvegicus) were obtained from Serum was separated in a refrigerated centrifuge at
Laboratory Animal Facility, CSIR-Indian Institute of approximately 5000 rpm for 10 minutes and analyzed using
Toxicology Research, Lucknow. The rats with body weight Rx Daytona (Randox) automatic analyzer.
ranging from 90-100 g were randomly selected for the
treatment. All rats, qualifying the veterinary health Pathology
examination, were acclimatized for 5 days before start of Necropsy
treatment. Total 160 Wistar rats were divided in to four
Rats from all groups in the study were subjected to
groups comprising 40 animals each (20 males and 20 females).
detailed necropsy and findings were recorded. The rats to
The four groups so formed consisted of one control and
be sacrificed at term were fasted overnight, weighed,
three treated groups.
Table 2. Haematological parameters observed in male rats after dietary exposure to the test item

Mean
Mean Mean
White blood Red blood Haemo- Haema- Corpuscular Neutro-
Corpus- Corpuscular Platelet Lymphocyte Monocyte Eosinophil Basophil
Group Corpuscles Corpuscles globin tocrit Haemoglobin phil
cular Value Haemoglobin (103/µL) (%) (%) (%) (%)
(103/µL) (106/µL) (g/dL) (%) Concentration (%)
(fL) (pg)
(g/dL)
13.91 9.72 15.20 45.22 46.58 15.67 33.63 966.17 25.43 66.69 4.88 2.79 0.18
Control ± ± ± ± ± ± ± ± ± ± ± ± ±
4.14 0.70 1.01 3.29 2.61 0.71 0.78 119.36 13.43 13.80 1.63 1.40 0.11
12.95 9.20 14.40 45.81 46.34 15.66 33.88 974.00 20.73 70.38 4.89 4.01 0.07
5000 ppm ± ± ± ± ± ± ± ± ± ± ± ± ±
3.09 0.81 1.10 12.07 2.62 0.58 1.03 263.16 6.77 7.95 1.29 2.06 0.05
14.69 9.15 14.47 43.24 47.31 15.82 33.47 952.33 16.27 75.32 4.09 4.20 0.11
10000 ppm ± ± ± ± ± ± ± ± ± ± ± ± ±
6.75 0.46 0.63 1.92 1.83 0.51 0.67 155.53 3.76 3.47 1.56 2.07 0.04
15.17 9.19 14.69 43.44 44.44 16.04 33.87 724.28 16.08 74.68 4.67 2.86 0.13
20000 ppm ± ± ± ± ± ± ± ± ± ± ± ± ±
5.25 0.75 1.11 2.71 10.84 0.41 0.78 271.14 5.83 9.84 1.31 1.04 0.09

(Mean ± SD)(Significant at the level of P < 0.05)

Table 3. Haematological parameters observed in female rats after dietary exposure to the test item
Mean
Mean Mean
White blood Red blood Haemo- Haema- Corpuscular Neutro-
Corpus- Corpuscular Platelet Lymphocyte Monocyte Eosinophil Basophil
Group Corpuscles Corpuscles globin tocrit Haemoglobin phil
cular Value Haemoglobin (103/µL) (%) (%) (%) (%)
(103/µL) (106/µL) (g/dL) (%) Concentration (%)
(fL) (pg)
(g/dL)
10.06 8.04 13.41 40.58 50.52 16.68 33.02 872.69 18.88 72.82 4.58 3.68 0.28
Control ± ± ± ± ± ± ± ± ± ± ± ± ±
3.41 0.75 1.18 3.36 1.98 0.57 0.64 122.85 4.01 4.01 1.90 1.08 0.33
9.65 8.25 14.03 42.44 51.50 17.02 33.06 849.06 18.59 70.33 5.42 5.58 0.12
5000 ppm ± ± ± ± ± ± ± ± ± ± ± ± ±
2.59 0.38 0.58 1.63 1.72 0.47 0.76 109.92 5.97 7.62 1.51 2.65 0.08
10.28 8.31 13.86 42.01 50.61 16.69 32.99 869.60 18.46 71.67 4.39 5.32 0.15
10000 ppm ± ± ± ± ± ± ± ± ± ± ± ± ±
3.93 0.48 0.75 2.31 1.75 0.33 0.65 151.97 4.66 5.30 1.38 2.60 0.13
10.32 8.40 14.05 42.59 50.68 16.73 32.98 924.46 16.78 74.65 4.79 3.62 0.12
20000 ppm ± ± ± ± ± ± ± ± ± ± ± ± ±
SRIVASTAVA AND THOMBARE, Safety Assessment of Shellac as Food Additive through Long Term Toxicity Study

2.96 0.39 0.61 1.67 1.40 0.43 0.44 168.10 4.39 5.23 1.01 1.10 0.06
(Mean ± SD)(Significant at the level of P < 0.05)
735
Table 4. Biochemical parameters observed in male rats after dietary exposure to the test item 736

Aspartate Alanine
Total Trigly- Alkaline Total
Glucose Creatinine amino Urea Albumin Uric Acid Cholesterol Globulin amino
Group Protein cerides phosphatase Bilirubin
(mg/dL) (mg/dL) Transferase (mg/dL) (g/dL) (mg/dL) (mg/dL) (mg/dL) transferase
(g/dL) (mg/dL) (U/L) ((mg/dL)
(U/L) (U/L)
50.75 0.50 120.82 29.33 5.23 59.57 2.34 0.79 45.18 2.84 28.18 176.45 0.07
Control ± ± ± ± ± ± ± ± ± ± ± ± ±
12.12 0.09 33.79 5.63 0.75 22.28 0.41 0.17 11.94 0.47 6.43 83.93 0.05
73.87 0.41 88.24 24.30 4.20 72.15 2.17 1.04 38.36 2.07 31.94 303.41 0.07
5000 ppm ± ± ± ± ± ± ± ± ± ± ± ± ±
25.81 0.17 62.13 7.76 1.43 33.40 0.71 0.41 13.00 0.78 14.04 175.19 0.05
101.63 0.57 154.95 27.51 7.62 115.04 2.84 2.17 68.13 4.71 46.95 186.42 0.04
10000 ppm ± ± ± ± ± ± ± ± ± ± ± ± ±
30.29 0.12 39.37 4.13 0.67 49.83 0.42 0.83 10.64 0.52 13.57 65.00 0.05
103.34 0.59 173.07 31.90 6.05 81.15 2.94 1.53 53.16 3.08 42.64 204.07 0.11
20000 ppm ± ± ± ± ± ± ± ± ± ± ± ± ±
29.16 0.16 50.24 7.90 1.11 31.33 0.53 0.40 10.17 0.90 16.68 73.41 0.07

(Mean ± SD)(Significant at the level of P < 0.05)

Table 5. Biochemicalparameters observed in female rats after dietary exposure to the test item
Aspartate Total Trigly- Alanine Total
Glucose Creatinine Urea Albumin Uric Acid Globulin Alkaline
amino Protein cerides Cholesterol amino Bilirubin
Group phosphatase
(mg/dL) (mg/dL) Transferase (mg/dL) (g/dL) (mg/dL) (mg/dL) (mg/dL) transferase
(g/dL) (mg/dL) (U/L) ((mg/dL)
Trends in Biosciences 10 (2), 2017

(U/L) (U/L)
89.06 0.55 123.67 28.95 6.28 206.01 3.45 1.13 61.97 2.82 32.00 154.82 0.15
Control ± ± ± ± ± ± ± ± ± ± ± ± ±
20.67 0.16 53.35 5.90 1.46 74.60 0.88 0.53 14.20 0.74 16.68 55.64 0.08
125.03 0.62 181.00 29.62 9.58 183.57 3.36 2.53 71.86 6.05 51.29 136.15 0.02
5000 ppm ± ± ± ± ± ± ± ± ± ± ± ± ±
36.91 0.07 75.99 4.33 1.97 65.28 0.85 1.10 19.76 1.87 12.63 45.28 0.04
87.26 0.53 128.20 33.48 7.50 100.90 2.95 1.77 58.24 4.55 39.80 137.89 0.06
10000 ppm ± ± ± ± ± ± ± ± ± ± ± ± ±
23.36 0.11 41.07 8.37 1.01 26.93 0.32 0.64 9.05 0.77 12.61 57.39 0.07
124.84 0.78 125.23 32.82 7.69 152.57 1.42 1.89 60.32 6.27 12.23 151.85 0.20
20000 ppm ± ± ± ± ± ± ± ± ± ± ± ± ±
33.66 0.07 21.37 5.58 0.46 58.57 0.74 0.54 10.33 0.71 4.32 67.49 0.00

(Mean ± SD)(Significant at the level of P < 0.05)

 SRIVASTAVA AND THOMBARE, Safety Assessment of Shellac as Food Additive through Long Term Toxicity Study
anaesthetized with thiopentone injection (intra-peritoneal)
and exsanguinated.
Tissue collection
On completion of the gross pathology examination
the tissues and organs were collected from all animals,
preserved in 10% formal saline and weighed.
Histopathology
Histopathological examination was carried out on the
preserved organs of control and high dose group rats. The
tissues were processed for routine paraffin embedding and
sections were stained with Haematoxylin and Eosin stain.
Unused tissues were archived.There were no gross lesions
in any of the rats that were subjected to a gross pathology
examination. Since no organ/tissue showed test item related
histopathological changes in the high dose groups as
compared to the control animals, tissues from the lower
dose groups were not examined.
Statistical analyses
The statistical analysis of the experimental data was
carried out andall quantitative variables like laboratory
investigations (haematology and clinical chemistry) were
subjected to one-way ANOVA test.All analysis and
comparisons were evaluated at the 5 % (P <0.05) level.
RESULTS AND DISCUSSION
Analysis of the test sample
As quality of shellac may differ from one batch to
another, due to effect of climate, host plant, crop, etc., sample
from single batch was used for carrying out the toxicological
study. Shellac used for the study was tested for its
composition and determined to contain 95.2% total resin,
4.8% wax and tracesof coloring components. Wax free
portion when further fractioned, gave approximately 74%
hard (ether insoluble) resin and 26% soft (ether soluble)
resin. Before undertaking toxicological studies, the sample
was tested for following physico-chemical parameters (IS
6921:1973), as shown in table 1.
Figure2 shows FTIR spectrum of handmade shellac.
The summary of bands position and their assignments for
each peak are set out (Limmatvapirat et al., 2008), which
can be summarized as follows:
A broad shoulder peak at 3100-3500 cm-1 belongs to
stretching of O-H group. Sharp peak at 2854 and 2920 cm-1
correspond to asymmetric stretching vibration of C-H
groups. A sharp peak at 1712 cm-1 represent stretching
vibration of -C=O due toester group. The prominent peak
at 1250 cm-1is attributed to C-O asymmetric stretching
vibration of fatty acid components from shellac.
Toxicity study of test sample
General observation and mortality
In case of clinical signs/behavior of animalsof the
control or test item treated group,no abnormality was
recorded during the course of the study. Since the mortalities
in the control or treated groups of animals were at par, hence
this was not attributed due to the test item administered.
737
There was no significant difference in body weight gain
between groups nor was there any significant loss in body
weight during the course of the study as indicated in figure
3. Body weight gain during the course of the study was
comparable in all animals of both control and treated groups.
Rats from all the groups exhibited similar feed consumption
trends throughout the course of study. The increase in
feed consumption with time was comparable in all the groups
as indicated in figure 4.
Clinical pathology investigations (hematology, clinical
chemistry)
Clinical pathology investigation, including
haematological and biochemical parameters were examined
separately for control and all the treated groups. Changes
brought about by the test item administration in the
haematological parameters of the male and female animals
are depicted in table 2&3 respectively. Slight reduction
ofplatelets in mid dose, male group by 1.5% was observed
which further enhanced in high dose group by 25%. No
such trend was observed in case of female groups. Overall
reduction in the neutrophils of animals in male group at all
three doses was observed. However the reduction was
substantially higher in mid and high dose group which was
36.0 and 36.7% respectively. Female rat group also showed
reduction in neutrophils at high dose by 11.1% as compared
to the control one.
Biochemical parameters were determined separately
for males and females, as discussed in table 4 and 5
respectively. Tremendous increase in glucose in mid dose,
male group was observed which was exactly doubled as
compared to the control one. This glucose level further
enhanced foranimals in high dose group. Remarkable
increase in the level of uric acid was recorded, especially in
the animals of mid dose group of male and female, which
was 174% and 56.6% respectively. Increase in glucose and
uric acid among females, only in low dose group was neither
biologically significant nor was dose dependent. Acute
increase in triglycerides of animals by 93% as compared to
control was observed in mid dose, male group. Whereas, in
case of mid dose female group level of triglycerides reduced
to 51%. Noteworthy increase in the level of cholesterol by
50% and total protein by 45.7% of the animals in mid dose
male group was detected, however no significant changes
were observed in case of low dose male and female groups.
Anomalous significant changes were observed in albumin
content in mid as well as high dose group of male and
female animals. In case of males, the value recorded was
higher by 21.3%, whereas for females it was lower by 14.5%
as compared to control one.
On account of the results of the haematological and
biochemical investigation, it was observed that most of the
parameters became significant from mid and high dose of
the test item. Variations in the values of various parameters
for male and female group were neither biologically
significant nor was dose dependent at low dose of shellac,
i.e. 5000 ppm in powdered feed. On close proximity of the
data analyzed, it was concluded that the shellac can be
administered to the test animals up to the dose of 5000 ppm
safely.
738 Trends in Biosciences 10 (2), 2017

Fig. 1. Structure of shellac resin Fig. 2. FT-IR spectrum of shellac

Pathology presented in the report. Organ / body weight ratios of vital

Necropsy
At necropsy, the animals were examined visually for
external abnormalities including palpable masses. The
abdominal, thoracic and cranial cavities and their contents
were examined for abnormalities. On completion of the gross
pathology examination, the tissues and organs like adrenal
glands, brain (including medulla/pons, cerebellum and
cerebrum), epididymides, heart, kidneys, liver, ovaries,
spleen, testes, uterus (with cervix), etc. were collected from
all animals weighed and preserved in 10% formal saline.
Paired organs were weighed together and combined weight
organs of treated rats did not differ significantly in
comparison with controls (Figure 5); except slight increase
in liver at 10000 ppm diet dose in female rats and minor
variation in few other organs which were found significant
at the level of P < 0.05. Gross pathology observations
revealed congested lungs in three rats (two females from
control and one male from low dose group) and a pale liver
in one male ratlow dose group.
Histopathological observations
Histopathological examinations carried out, on the
preserved organs of male and female rats and the

Fig. 3. Body weight gained by the male and female Wistar rats in course of study (180 days)

Fig. 4. Feed consumption by the male and female Wistar rats in course of study (180 days)
 SRIVASTAVA AND THOMBARE, Safety Assessment of Shellac as Food Additive through Long Term Toxicity Study
Fig. 5. Effect of long term (26 weeks) dietary exposure of lac dye on mean organ body weight ratio (%) of male and
femalerats at necropsy
[ Organ body weight ratio= (Organ weight x 100)/ Body weight ]
observations taken for animals from control and high dose
groups are explained as follows.
Control Group: Lungs from one male animal showed
peribronchiolar lymphoid hyperplasia. Adrenal from two
male animals showed congestion. Liver from two male
animals revealed congestion and foci of mononuclear cells
(MNCs) accumulation respectively. Spleen from one male
showed lymphoid hyperplasia. Kidney from one male
depicted foci of MNCs and one female showed congestion.
Uterus from one animal revealed mild hyperplasia of
endometrial glands. Ovary from one animal showed mild
congestion.
High dose group: Lungs from high dose treated group
animals showed lesions like Peribronchiolar lymphoid
hyperplasia (one male), congestion (one male) and alveolar
consolidation (one female). Liver showed lesions like foci
of MNCs (one male) and congested central veins (one
female). Spleen showed lesions like congestion (one female)
and lymphoid hyperplasia (one female). Congestion was
observed in the adrenal (one female) and kidney (one male).
Ovary from one animal revealed congestion, whereas testis
from one animal depicted oligopsermia.
Lesions observed in high dose group appear to be
spontaneous and incidental in nature also they are
comparable with control hence cannot be related with the
treatment. There were no gross lesions in any of the rats
that were subjected to a gross pathology examination. Since
no organ/tissue showed test item related histopathological
changes in the high dose groups as compared to the control
animals, tissues from the lower dose groups were not
examined.
Long term toxicity studies to assess the systematic
toxicological effect of the shellac revealed that, food intake
was comparable in control as well as test item administered
rats at all dose levels. None of the animals in low, mid and
high treatment groups exhibited any statistically significant
abnormal clinical sign or behavioral change during the
course of study. There were no treatment-related gross and
microscopic changes in both male and female rats of the
high dose group when compared to the rats from the control
group. Significant impact of shellac was observed on
various haematological and biochemical parameters like,
739
platelets, neutrophils, glucose, uric acid, triglycerides,
cholesterol, total protein and albumin at mid and high dose
animal groups.But, no significant variations in the values
of haematological and biochemical parameters for male and
female group were observed at low dose of shellac, i.e. 5000
ppm in powdered feed. It is therefore, concluded that
repeated 180 days dietary exposure of shellac up to the
dose of 5000 ppm through feed did not produce any
significant toxic effects in male and female rats.
ACKNOWLEDGEMENT
The authors express their sincere gratitude to Indian
Council of Agricultural Research, for providing financial
support in carrying out this study. The authors thankfully
acknowledge Dr. Akshay Dwarkanath, and Dr. B D
Bhattacharjee, CSIR-Indian Institute of Toxicological
Research, Lucknow,for facilitating this study.
LITERATURE CITED
Baboo, B. and Goswami, D. N. 2010. Lac: an introduction. In:
Processing, Chemistry and Applications of Lac, Directorate of
Information and Publications of Agriculture, Indian Council of
Agricultural Research, New Delhi, pp. 1-6.
Bose, P. K., Shankarnarayanan, Y. and Sengupta, S. C. 1963. Physical
properties and chemical constants. In: Chemistry of Lac, Indian
Lac Research Institute, Ranchi, India, p. 19.
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Received on 18-01-2017 Accepted on 23-01-2017