ISSN (Print) : 0974-6846

Indian Journal of Science and Technology, Vol 9S(1), DOI: 10.17485/ijst/2016/v9iS1/109826, December 2016 ISSN (Online) : 0974-5645

A Study on the Whitening Effect of Fruit Extracts and

the Changes in the Components of the Teeth
Jung-Hyun Lee and Sung-Suk Bae*
Department Dental Hygiene/ Hanseo University, Korea;
[email protected], [email protected]

Abstract
Background/Objectives: In this regard, this study aimed to examine the teeth-whitening effect of undiluted and diluted
extracts of Korean natural citron, grapefruit, and plum and analyze the components of teeth. Methods/Statistical Analysis:
In this study, 55 premolars in the upper and lower jaw extracted in recent few months were used as test pieces. Extracts
of citron, grapefruit, and plum were used as solutions. Each of the test pieces was deposited into undiluted solution and
diluted solution (one to one ratio of undiluted solution in distilled water) for 3, 6, and 9 hours, respectively. The color
difference was examined using a Spectrophotometer (CM-3500d, Minolta, Japan). Changes in components of the teeth
after they were deposited into the extracts of citron, grapefruit, and plum were compared to their initial state. SPSS ver.
20.0 was used for statistical analysis. Mann-Whitney test was performed to compare the pre-post difference within and
between test pieces. Kruskal-Wallis test was implemented to compare the averaged values of E* (color difference) by time.
Tukey´s test was carried out with 95% confidence interval for the post test. Findings: The whitening effect of citron extract
was increased as time went by. The diluted solution with one to one ratio of the citron extract in distilled water showed
the highest whitening effect after depositing for 3 hours. The whitening effect of grapefruit extract was increased as time
went by. The diluted solution with one to one ratio of the grapefruit extract in distilled water showed the highest whitening
effect after depositing for 6 hours. Plume extract had the highest whitening effect after depositing for 6 hours. The diluted
solution with one to one ratio of the plum extract in distilled water showed the highest whitening effect after depositing
for 3 hours. Components of P and Ca were irregularly increased or decreased in test pieces both before and after the
depositions of teeth into fruit extracts. Improvements/Applications: In this study, the colors of teeth in extracts of citron,
grapefruit, and plum turned brighter than the initial color as time went by. Follow-up studies are needed.

Keywords: Citron, Grapefruit, Natural Liquid Fruit, Teeth-whitening, Teeth Whitening Effect

1.  Introduction matters and water (4%). Mineral crystals are opaque with
organic proteins present between them. Consequently,
Dental bleachers currently used in the dental clinics pigments may soak into this space.2 Tooth whitening has
contain carbamided peroxide, sodium perborate, and been consistently implemented since its introduction.
hydrogen peroxide that might cause side effects such as Various kinds of dental bleachers have been developed,
root resorption, dentin sensitivity, and stomatitis.1 Tooth produced, and used in many countries. In South Korea,
enamel is composed of minerals (96%) and organic various kinds of dental bleachers are being imported in

*Author for correspondence

 A Study on the Whitening Effect of Fruit Extracts and the Changes in the Components of the Teeth

various ways such as expert bleaching and home bleach- toothpaste. They found significant difference at,4 weeks
ing3. In addition, recently, Over-The Counter (OTC) after using the tooth-whitening toothpaste with significant
bleachers have been popularized. They can be used with- color change. Lee 2l has reported that carbamide peroxide
out prescriptions from experts. As representative OTC and hydrogen peroxide are not carcinogenic. However,
bleachers, whitening toothpaste and patches containing they might stimulate chemical carcinogenicity. Ham1 and
hydroxyapatite and peroxide compounds are commonly Gang10 have reported that the use of highly concentrated
used4. However, chemically synthesized dental bleach- carbamide peroxide, one of the main components in den-
ers might have side effects if are wrongfully used. Under tal bleachers, might cause dentin hypersensitivity and
these circumstances, more emphasis has been recently changes in enamel and gingiva as well as excessive saliva.
put on the assurance of their stability for the human body. In other words, the whitening effect of safe and natural
There has been mounting interest in alternative whiten- dental breachers (i.e., the extracts from citron, grapefruit,
ing using vegetables and fruits as sources of natural dental and plum) can significantly increase the values of E*.
bleachers. As a result, the assurance of their stability for There is a need to perform further experimental research
the human body has recently become important and to find natural materials for the development of natural
more interest arises in the alternative whitening using dental bleachers.
vegetables or fruits as natural dental bleachers. Studies As the result of analysis of the enamel surface of the
on whitening with natural materials are on the rise. The deposited test pieces before and after they were deposited
whitening effect of citron has been identified in previous in fruit extracts, it was found that the ratio of the values
studies. It is currently used for skin care.5 The extract of of P and Ca were decreased compared to those before
grapefruit has been found to have antimicrobial effect.6,7 the deposition, while the opposite was true for grapefruit
The extract from plum has been reported to have function extract. Kim25 has performed component analysis using a
of depressing the growth of melanin.8 In addition, its anti- scanning electron microscope and reported that the val-
microbial effect is increased as time goes by and when its ues of P and Ca of the teeth are decreased as the acidity
concentration is increased.9 Citron, grapefruit, and plum of the drink deposited for 15, 30, 60, and 120 minutes is
extracts are Koreans’ favorite fruit extracts. They are con- intensified with the passage of time. Lee26 has found that
sumed in various ways. In this regard, this study aimed to enamel erosion has occurred in the anterior teeth depos-
provide basic data for developing natural dental bleachers. ited in orange juice with pH level of 4 for 10 minutes. In
To meet this purpose, this study deposited extracted teeth addition, the micro-hardness value was decreased, even-
pieces into Korean natural citron, grapefruit, and plum tually causing an erosion on the enamel surface. Other
extracts to determine their whitening effects. In addition, research findings25 has implied that soft drinks if frequently
the components of the teeth before and after bleaching by consumed can also influence the calcium and phosphorus
the three fruit extracts were analyzed. contents of the enamel. Tooth deposition analysis of teeth
Recent research on whitening has been performed in deposited in natural fruit extracts also provided the same
various ways, including expert dental bleaching and com- implication. Meanwhile, Kim et al.27 have found that if the
ponents of bleachers.1–4,10–23 As the result of analysis of teeth are deposited in a drink with Ca added, the damage
color difference in this study, it was found that the values to the surface of enamel will be reduced. In this regard, a
of E* of tooth piece was increased after it was deposited careful examination of the decrease in P and Ca contents is
in the extracts of citron, grapefruit, and plum compared needed based on the results of previous studies. Additional
to the initial value of the test piece, demonstrating that studies are needed to verify the results of this study.
these fruit extracts have a statistically significant whit-
ening effect (P< 0.05). Jung et al.24 have also examined 2.  Proposed Work
the teeth-whitening effect of several species of medicinal
herbs. They deposited carious teeth in their extracts for Step 1: Producing test pieces
16 hours and verified their whitening effects based on the Teeth pieces for the experiment were premolars of
fact that test pieces turned brighter compared to their ini- the upper and lower jaws extracted due to orthodontic
tial color. Meanwhile, Kim et al.17 have used chemically procedures or periodontitis. They were healthy without
synthesized whitening tooth paste for 3 weeks and deter- history of enamel hypoplasia, dentin exposure, or cavity
mined its tooth-whitening effect compared to a general treatment. This study prepared a total of 55 devitalized

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 Jung-Hyun Lee and Sung-Suk Bae

teeth pieces, including 1 untreated tooth and 54 pieces measurement values were obtaining for the values of E*
assigned to citron, grapefruit, and plum extract (18 pieces after identifying the values of L, a*, and b, using Spectra
per extract). Tissues, residues, tartars, and exogenous magic program of the spectrophotometer connected to a
paint attached to the prepared teeth were removed with computer. The program could mathematically transform
an ultrasound scaler and a curette to scrap off periodontal the tristimulus color. Each of the test pieces was measured
tissues. These teeth were polished with pumice contain- from the center twice. The formula of the values of E* was
ing no glycerin, deposited in PerasafeⓇ solution for 30 expressed as follows.
minutes for sterilization, washed with an ultrasound
E* = {(ΔL*)2 +(Δa*)2 +(Δb*)2}1/2
washer, and immersed in sterile distilled water. A total of
55 moulds were produced to facilitate forming the molds where, E* was the value of color change of the test
of the test pieces by cutting a round plastic pipe (diam- pieces, L* was the value of the brightness of the test pieces
eter: 38 mm; height: 12 mm) with a high speed cutter. - the brightness of the object, 0 (black) to 100 (white), a*
Resin-clay which was easy to form molds without the was the chroma of the test piece - the value of the degree
need of high pressure was used to fill the moulds and fix from red to green color (the closer to 0, the closer to white
the extracted teeth. color ), b* was the chroma of the test piece - the values of
Step 2: Solutions the degree from yellow to blue color (the closer to 0, the
Extracts of citron, grapefruit, and plum grown in closer to white color ).
South Korea and their diluted extracts (with one to one Step 4: Observation through a scanning electron micro-
ratio of extracts vs. distilled water) were prepared as scope
shown in Table 1. Each extract was put into a 30 ml exper- Among test pieces deposited in the extracts of citron,
imental plastic beaker for 3, 6, and 9 hours. Meanwhile, grapefruit, and plum that were examined by a spectro-
diluted extracts were produced by mixing the extracts photometer, those with averaged values of L* and E* being
(15 ml) with distilled water (15 ml) at one to one ratio. the highest were chosen for scanning electron microscope
Afterwards, each dilution was put into an experimental (SEM) analysis. Using a scanning electron microscope
plastic beaker for 3, 6, and 9 hours. The extracted teeth (JEOL JSM-5600, Japan), the surface of the enamel of the
(n = 18) were put into the prepared beakers (3 in each test piece was examined by a factor of 16. Their compo-
beaker) and deposited in the extract or diluted extract. nents were analyzed to identify the difference from their
The teeth pieces were stored at temperature of 37℃ for 3, initial status.
6, and 9 hours and dried in a Drying Oven (C-DO, Korea)
to simulate oral condition after the deposition. Step 5: Statistical analysis
SPSS ver. 20.0 (SPSS Inc. Chicago) was used to analyze
whitening effect and changes in components. Samples
Table 1.  Bleaching solution used in this study
used in the experiment were utilized in a non-parametric
Product Brand Manufacturer test. Mann-Whitney test was performed to effectively ver-
name ify the pre-post difference within and between test pieces.
citron concentrate ES Food Young Woo Food, Korea Kruskal-Wallis test was performed to compare the aver-
aged values of E* by deposition period. Tukey´s test was
Grapefruit ES OEM Production,
ES Food implemented with a 95% confidence interval for the post
concentrate Korea
test to identify the averaged difference in the values of E*
Chinese plum
ES Food Samjin Food, Korea of each group. Statistical significance level was set at 0.05.
concentrate
Step 6: Ethical consideration
This study was an experimental study on extracted
Step 3: Whitening measurement teeth using human materials. It was performed after receiv-
To measure whitening, a spectrophotometer ing the approval from the IRB of the National Bioethics
(CM-3500d, Minolta, Japan) was used to measure the Committee regarding the research ethics. The study was
initial colors of the test pieces and the final colors after exempted from deliberation (Project Management No. :
being deposited in fruit extracts. The spectrophotometer P01-201511 -31-002).

Vol 9 S(1) | December 2016 | www.indjst.org Indian Journal of Science and Technology 3
 A Study on the Whitening Effect of Fruit Extracts and the Changes in the Components of the Teeth

3.  Results hours. In the diluted extract of plum extract (one to one
dilution of plum extract in distilled water), the highest
Step 1: Color difference of the test pieces deposited in whitening effect was found for test piece deposited in the
fruit extracts diluted extract for 3 hours followed by that after deposi-
Results of test pieces deposited in each fruit extract tion for 6 and 9 hours (P< 0.05).
and examined with the spectrophotometer are summa-
rized in Table 2. The values of E* of test pieces deposited Step 2: Changes in components before and after depo-
in citron and grapefruit extracts were found to have sig- sition
nificantly higher whitening effect as time went by. The A scanning electron microscope (JEOL JSM-5600)
highest whitening effect was found in plum extract after was used to examine the changes in components of teeth
depositing for 6 hours followed by deposition in plum before and after the test pieces were deposited in fruit
extract for 9 and 3 hours (P< 0.05). For the diluted extract extracts for 6 hours to determine the difference before
of citron (one to one ratio of distilled water and citron and after the deposition. Results of the measurement are
extract), the higher whitening effect was obtained for test shown in Table 3.
piece deposited in the diluted extract for 3 hours, followed The ratios of P and Ca of the test pieces before their
by that after deposition for 9 and 6 hours. In the diluted deposition were 10.85 and 20.43, respectively. After deposi-
extract of grapefruit extract (one to one ratio of distilled tion in the citron extract for 6 hours, the ratio of P (9.64) and
water and grapefruit extract), the highest whitening effect Ca (17.15) were both reduced. The same was true for test
was found for test piece deposited in the diluted extract pieces deposited in diluted citron extract, with the ratios of
for 6 hours, followed by that after deposition for 9 and 3 P and Ca of 10.42 and Ca 20.30, respectively. However, after

Table 2.  Color change of L* and E* values after settling in extract over time
Division Time N L* E* P
Before After
citron 3h 3 69.10 71.30 6.15a .000
an undiluted solution 6h 3 69.81 73.00 8.17 b

9h 3 68.53 71.68 10.03b

Distilled water diluent(1:1) 3h 3 67.38 70.65 7.35b .000
6h 3 68.68 71.60 6.30 a

9h 3 67.46 70.83 7.11b

Grapefruit 3h 3 65.74 66.93 1.86a .000
an undiluted solution 6h 3 66.69 68.10 2.02 a

9h 3 65.42 66.60 2.91b

Distilled water diluent(1:1) 3h 3 68.18 70.29 2.49a .000
6h 3 69.02 70.14 4.12 b

9h 3 71.02 72.58 3.62a

Chinese plum 3h 3 63.92 65.15 1.56a .000
an undiluted solution 6h 3 66.55 67.17 2.52b
9h 3 69.03 70.48 2.17b
Distilled water diluent(1:1) 3h 3 68.36 71.13 5.52a .000
6h 3 67.35 70.69 7.45 b

9h 3 64.45 66.67 5.32a

Values are reported as the mean
P-values were determined by Kruskal-Wallis test
a.b. means followed by different letters are significantly different

4 Vol 9 S(1) | December 2016 | www.indjst.org Indian Journal of Science and Technology
 Jung-Hyun Lee and Sung-Suk Bae

deposition in grapefruit extract, the ratios of P and Ca were 2. Based on the values of E*, grapefruit extract had sig-
increased to 13.27 and 24.90, respectively. After deposition nificant whitening effect as time went by. In diluted
in the diluted grapefruit extract, the ratio of P was increased solution of grapefruit extract, the whitening effect was
to 12.10 but the ratio of Ca was decreased to 18.99. After the highest (P< 0.05) after 6 hours of deposition.
deposition in the plum extract, the ratios of P and Ca were 3. The whitening effect of the plum extract was the high-
decreased to 10.31 and 18.31, respectively. However, after est after deposition of test piece for 6 hours, followed
deposition in the diluted plum extract, the ratios of P and Ca by that for 9 and 3 hours of deposition. In diluted
were increased to 11.88 and 21.56, respectively. plum extract, the whitening effect was the greatest (P<
0.05) after 3 hours of deposition.
4.  Conclusion 4. Component analysis of the three fruit extracts before
and after deposition showed that the ratios of P (9.64)
This study identified a meaningful difference in the whit- and Ca (17.15) of test pieces deposited in citron
ening effect of fruit extracts and component changes of the extract were decreased compared to their initial val-
teeth after deposition in fruit extracts of citron, plum, and ues. The same was found for the ratios of P (10.31) and
grapefruit. A total of 55 extracted teeth were used to produce Ca (18.31) of test pieces deposited in plum extract.
the test pieces to examine their color changes depending on However, for test pieces deposited in grapefruit
the duration of their deposition in natural fruit extracts pro- extract, the ratios of P (13.27) and Ca (24.90) were
duced in South Korea. The results of this study are as follows: increased compared to their initial values.
1. Based on the values of E* of test pieces, citron extract This study found that natural fruit extracts of citron,
was found to have significant whitening effect as time grapefruit, and plum had teeth-whitening effect. This may
went by. In the diluted solution at one to one ratio of have important implication for the development of nat-
the extract with distilled water, the whitening effect of ural dental bleachers. However, changes in components
the dilution extract was found to be the highest after 3 of teeth such as P and Ca need to be closely examined.
hours of deposition. The limitations of this study are as follows. First, this

Table 3.  Results of elements analyses (Normalized)

Division C O Na Al Si P Cl Ca Sn Sb I W %

Incipient 33.72 28.24 0.17 0.11 0.34 10.85 0.34 20.43 0.77 3.11 1.09 0.84 100
(Untreated)
citron 39.51 26.20 0.18 0.11 1.30 9.64 0.32 17.15 0.74 3.10 0.97 0.75 100
an undiluted
solution
Distilled 34.47 27.05 0.17 0.12 1.05 10.42 0.38 20.30 0.78 3.33 1.12 0.79 100
water
diluent(1:1)
Grapefruit 19.43 35.01 0.26 . 0.45 13.27 0.53 24.90 0.86 3.22 1.15 0.94 100
an undiluted
solution
Distilled 18.53 44.04 0.35 . 0.40 12.10 0.38 18.99 0.63 2.80 0.93 0.86 100
water
diluent(1:1)
Chinese plum 34.31 28.10 0.21 0.11 0.92 10.31 0.30 18.31 1.01 4.42 1.36 0.64 100
an undiluted
solution
Distilled 25.73 31.99 0.24 0.10 0.57 11.88 0.39 21.56 1.04 4.48 1.31 0.73 100
water
diluent(1:1)
All results in weight %

Vol 9 S(1) | December 2016 | www.indjst.org Indian Journal of Science and Technology 5
 A Study on the Whitening Effect of Fruit Extracts and the Changes in the Components of the Teeth

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