Comparative Evaluation of Salivary pH with Honey and Vinegar Mouth Rinse in Diabetic and Healthy Adults

Abstract

Background: Salivary fluid creates a particular environment of oral cavity that helps in mastication, lubrication of food and mucosa and in speech. Intake of food and different liquids (drinks, juices, milk) causes modulation in pH of saliva that lead to change in the environment of oral cavity. The pH of saliva decreases to acidic side when bacteria breakdown the carbohydrates and start releasing acids, these acids damage the structure of tooth and leads to cavity formation i.e. dental caries. Objective: The current study is aimed to evaluate the salivary pH of diabetic and healthy individual before and after using honey and vinegar mouth rinses. Methods: It was a pre-clinical experimental study conducted in dental OPD of Baqai medical college Karachi from 1st January to 15th February. The calculated sample size N=80 was divided in 4 groups, Group A, n=20 healthy participants who rinsed with honey mouth rinse, Group B n=20 diabetics patients who rinsed with honey mouth rinse. Similarly, Group C, n=20 healthy participants who rinsed with vinegar mouth rinse and Group D, n=20 diabetic patients who rinsed with vinegar mouth rinse. 2 ml of saliva was collected by asking the participants to collect it in the floor of the mouth and swallowing for one minute was prohibited and after that they were asked to expectorate it into the sterile container. Then they were given the mouth rinse according to group distribution and after rinsing they were asked to wait for half an hour after that saliva was again collected from same participant to identify the rinse induced change in pH of oral cavity. Results: There was no any significant change in healthy participants of either group however in diabetic individuals significant change was observed by honey mouth rinse (p-value = 0.033) followed by vinegar mouth rinse (p-value = 0.043). Conclusion: Honey and vinegar mouth rinses are effective in maintaining the salivary pH in diabetic individuals.

Full text

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*Corresponding author: E-mail: akhtar.ali@zu.edu.pk;
Journal of Pharmaceutical Research International
33(26B): 54-59, 2021; Article no.JPRI.67542
ISSN: 2456-9119
(Past name: British Journal of Pharmaceutical Research, Past ISSN: 2231-2919,
NLM ID: 101631759)
Comparative Evaluation of Salivary pH with Honey
and Vinegar Mouth Rinse in Diabetic and Healthy
Adults
Akhtar Ali
1*
, Lubna Farooq
2
, Anum Mahmood
3
, Shaikh Nadeem Ahmed
2
,
Asif Ahmed
2
and Sumreen Mujahid
2
1
Department of Pharmacology, Ziauddin University,
Pakistan.
2
Department of Pharmacology, Baqai Medical University, Karachi, Pakistan.
3
Department of Dental Materials, Dow University of Health Sciences Karachi, Pakistan.
Authors’ contributions
This work was carried out in collaboration among all authors. The concept of study, drafting and
finalizing of the results were done by author LF. Data analysis was performed by author AA. The
article was critically reviewed and finally drafted by authors AA and LF. Finally reviewed and approved
by author SNA. Laboratory/instrument based work was performed under the supervision of author AA
and assisted by authors SM and AM. All authors read and approved the final manuscript.
Article Information
DOI: 10.9734/JPRI/2021/v33i26B31482
Editor(s):
(1) Dr. Juan Carlos Troiano, University of Buenos Aires, Argentina.
Reviewers:
(1) Bashir Matata, Liverpool Heart & Chest Hospital and University of Liverpool, UK.
(2)
Marlon Bruno Nunes Ribeiro, Universidade Federal de Minas Gerais, Brasil.
Complete Peer review History:
http://www.sdiarticle4.com/review-history/67542
Received 23 March 2021
Accepted 23 April 2021
Published 28 April 2021
ABSTRACT
Background:
Salivary fluid creates a particular environment of oral cavity that helps in mastication,
lubrication of food and mucosa and in speech. Intake of food and different liquids (drinks, juices,
milk) causes modulation in pH of saliva that lead to change in the environment of oral cavity. The
pH of saliva decreases to acidic side when bacteria breakdown the carbohydrates and start
releasing acids, these acids damage the structure of tooth and leads to cavity formation i.e. dental
caries.
Objective: The current study is aimed to evaluate the salivary pH of diabetic and healthy individual
before and after using honey and vinegar mouth rinses.
Methods: It was a pre-clinical experimental study conducted in dental OPD of Baqai medical
college Karachi from 1
st
January to 15
th
February. The calculated sample size N=80 was divided in
4 groups, Group A, n=20 healthy participants who rinsed with honey mouth rinse, Group B n=20
Original Research Article

Ali et al.; JPRI, 33(26B): 54-59, 2021; Article no.JPRI.67542
55
diabetics patients who rinsed with honey mouth rinse. Similarly, Group C, n=20 healthy participants
who rinsed with vinegar mouth rinse and Group D, n=20 diabetic patients who rinsed with vinegar
mouth rinse. 2 ml of saliva was collected by asking the participants to collect it in the floor of the
mouth and swallowing for one minute was prohibited and after that they were asked to expectorate
it into the sterile container. Then they were given the mouth rinse according to group distribution
and after rinsing they were asked to wait for half an hour after that saliva was again collected from
same participant to identify the rinse induced change in pH of oral cavity.
Results: There was no any significant change in healthy participants of either group however in
diabetic individuals significant change was observed by honey mouth rinse (p-value = 0.033)
followed by vinegar mouth rinse (p-value = 0.043).
Conclusion: Honey and vinegar mouth rinses are effective in maintaining the salivary pH in
diabetic individuals.
Keywords: Honey; vinegar; rinses; diabetic individuals; salivary pH.
1. INTRODUCTION
Salivary fluid creates a particular environment of
oral cavity that helps in mastication, lubrication of
food and mucosa and in speech [1,2]. Major and
minor glands generally secret 0.5 to 1.5 litre
saliva in a day, and 6.2 – 7.2 pH is considered as
normal pH of saliva [3]. Intake of food and
different liquids (drinks, juices, milk) causes
modulation in pH of saliva that lead to change in
the environment of oral cavity. The pH of saliva
decreases to acidic side when bacteria
breakdown the carbohydrates and start releasing
acids, these acids damage the structure of tooth
and leads to cavity formation i.e. dental caries
[4]. Alkaline environment of saliva appears when
pH of saliva increases which causes
development of plaque and calculus. The health
of oral cavity and periodontal condition in neutral
pH has less chances of decay and periodontal
problems [5].
Diabetes mellitus is a health challenge faced by
the medical professionals worldwide, according
to a survey, 19.2% population of Pakistan has
Type II diabetes mellitus [6]. Being a part of
metabolic syndrome diabetes mellitus affects all
body systems including oral cavity. It contributes
to various oral ailments such as halitosis, caries,
gingivitis, periodontitis and ultimately loosening
of teeth by altering the salivary pH. This
alteration in salivary pH is also contributed by
decreased salivary rate as it is observed that in
diabetes mellitus due to systemic dehydration
and increase in salivary glucose patient may
complain of dry mouth which is also considered
as a risk factor for above mentioned problems
related to oral cavity [7]. Patients with
uncontrolled diabetic profile have low salivary
efficacy to neutralize the acidic environment of
oral cavity hence are more susceptible to
develop dental problems particularly dental
carries. Systemic hyperglycemia and
increase in amount of glucose in saliva facilitates
the growth of acidogenic bacteria i.e.
Streptococcus mutans, which ultimately leads to
tooth decay [8].
Many natural products such as medicinal herbs,
extract of seeds, flower, leaf and stem of different
plants as well as honey is also considered as a
good choice to prevent the dental problems [9].
Honey is a natural sweet product processed by
the honey bees, it is reported that honey is less
likely to participate in carries progression when
compared with glucose or fructose along with
that it is also highlighted that it has anti-bacterial,
anti-inflammatory, anti-ulcer and antioxidant
properties [10]. Due to its wide range properties it
has been found to use it as treatment of
gingivitis, bleeding and receding gums. Its
antimicrobial properties are attributed to enzyme
glucose oxidase which produces hydrogen per
oxide on mixing the honey with water [11].
Vinegar is made from fermentable sugar source
like dates, apple, grapes etc [12]. Studies have
reported its bactericidal effects and biofilm
removal properties that are beneficial for dental
health. Various studies has shown that after
washing toothbrushes with vinegar decreases
the number of streptococcus pyogens [13,14].
The current study is aimed to evaluate the
salivary pH of diabetic and healthy individual
before and after using honey and vinegar mouth
rinses.
2. MATERIALS AND METHODS
It was a pre-clinical experimental study
conducted in dental OPD of Baqai medical
college Karachi from 1
st
January to 15
th
February
2021. The sample size was calculated by sealed

Ali et al.; JPRI, 33(26B): 54-59, 2021; Article no.JPRI.67542
56
envelope software i.e. N=80. The participants
were divided into 4 groups by simple random
sampling technique. Patients with type II
diabetes (without other comorbidities) and
healthy individuals who visited the dental OPD
for regular visit were included as study
participants. Individuals with other oral and
systemic comorbidities were excluded. Grouping
was performed as; Group A, n=20 healthy
participants who rinsed with honey mouth rinse,
Group B n=20 patients with diabetes who rinsed
with honey mouth rinse. Similarly, Group C, n=20
healthy participants who rinsed with vinegar
mouth rinse and Group D, n=20 patients with
diabetes who rinsed with vinegar mouth rinse.
We enrolled participants who agreed and were
according to inclusion criteria and asked
educated them to maintain the oral hygiene for
15 days. They were instructed to refrain from
eating for 1 h before collection of saliva. For the
patients with diabetes Random Blood Sugar
levels were taken before the mouth rinse. 2 ml of
saliva was collected by asking the participants to
collect it in the floor of the mouth and swallowing
for one minute was prohibited and after that they
were asked to expectorate it into the sterile
container. Then they were given the
mouth rinse according to group distribution and
after rinsing they were asked to wait for half an
hour after that saliva was again collected from
same participant to identify the rinse induced
change in pH of oral cavity. pH strips were
immersed in the sterile container and reading
were noted. It was reassured by the
examiner that the pH strips were uncontaminated
and were kept in place for 30 seconds and used
as per manufacturer’s guidelines. The colour
change was noted and readings were compared
with the indicator chart given in the pH strip
packing.
2.1 Preparation of Mouth Rinse
Ten ml of commercial honey was added to 100
ml of distilled water, and mixed for 10 minutes
until it was homogenized. Same protocol was
followed for the vinegar mouth rinse and was
kept at room temperature in sterile bottle until
use. Both the solutions were reduced to 10 ml to
obtain 100% (W/V) concentration [15]. Prior to
experiment A pilot study was conducted on 6
participants to determine the acceptability and
safety of the mouthwashes and no adverse
events were reported.
The data collected was analyzed using SPSS
version 21 for descriptive and analytical
statistics. The interpretations of the pH readings
are based on the colour coding chart. pH
between the range of 4.0–5.5 shows it to be in
the acidic range which can be improved by the
change in the dietary pattern to benefit the pH of
one’s body. The pH reading in between 6.5 and
7.5 is the optimal range to maintain the alkalizing
lifestyle. Above 8 is uncommon showing
salivary pH to be too alkaline which is not
normal. Paired T test was applied to record the
pre and post rinsed oral pH followed by ANOVA
for inter group comparison of post rinsed pH
values with p-value less than 0.05 considered as
significant.
3. RESULTS
The mean age of patients with diabetes (group B
and D) was 53.4 ± 8.2 years and mean age of
healthy individuals (Group A and C) was 45.9 ±
9.6 years. There were 56 female and 24 male
participants in our study. The random blood
sugar levels in patients with diabetes were 203.5
± 23.4 mg/dl
3
. In group A, there was no
significant change in pH of saliva before and
after rinsing with honey mouth rinse, however,
when Group B i.e., diabetic individuals were
checked significant increase in saliva was
observed after 30 of rinsing as shown in Table 1.
Table 1. The observed pH of group A and B
before and after honey mouth rinse
pH before
honey rinse
pH after
honey rinse
p-
value
Group A
5.95 ± 1.8
6.1 ± 1.02
0.133
Group B 5.01 ± 1.49 6.75 ±0.58 0.033
Table 2. The observed pH of group C and D
before and after vinegar mouth rinse
pH before
vinegar
rinse
pH after
vinegar
rinse
p-value
Group C
5.56 ± 1.02
5.90±1.49
0.142
Group D
5.42 ± 1.83 6.1 ± 1.0 0.043
Table 3. ANOVA analysis after rinsing
Groups pH after rinsing p-value
Group A 6.1 ± 1.02
0.024 Group B 6.75 ± 0.58
Group C 5.90 ± 1.49
Group D 6.1 ± 1.0

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The analysis of Group C showed no significant
difference before and after rinsing with vinegar
mouth rinse while analysis of Group D showed
significant increase in pH of saliva of
diabetic individuals results are highlighted in in
Table 2.
The intra group comparison of mouth rinses
showed that honey mouth rinse was significantly
more effective than vinegar mouth rinse in
increasing the salivary pH of oral cavity in
diabetic individuals.
4. DISCUSSION
Diabetes mellitus is a chronic iceberg disease
and public health problem which effects the
individuals of any age. Patients suffering from
diabetes compromise the salivary gland function
and results in decrease alteration of salivary flow
[16,17]. Saliva contains a variety of host defence
factors. Decrease in salivary flow may manifest
to oral complications such as xerostomia which
manifest as dental caries, gingivitis, periodontitis,
and certain fungal infections [18]. According to
WHO statistics about 80% of the population is
using natural products due to their potent
activates and less disadvantages. Natural
products now becoming popular approach,
especially in developing countries The
antibacterial activity of honey was known
because it contain methylglyoxal and presence
saturated groups in honey [19]. Without dilution
original form of honey was used in various in
vitro and in vivo studies against dental plaque
and caries [20], but the current study was
Planned to explore the effectiveness of diluted
form of honey as mouth rinse in diabetic and
non-diabetic patients. Honey contains many
enzymes such as hydrogen peroxidase when
diluted it become potent antibacterial rinse. In our
study when healthy individuals were assessed
for honey and vinegar mouth rinse there was no
any significant change observed before and after
the rinse, it is observed in multiple studies that
diabetes mellitus is one of the reasons that
causes decrease in salivary pH which results in
dental problems and increase the susceptibility of
affected individuals towards oral infections [21,
22]. While when it comes to healthy individuals
there is no significant change in pH of saliva
which is showing the parallel findings with above
studies as well as highlighting the protective
effects of saliva in healthy individual. It was
observed in diabetic individuals that their salivary
pH before the rinse was slightly acidic when
compared to healthy individuals which provides a
good media for growth of acidogenic bacteria
such as Strep. mutans that leads to more
pronounced dental and periodontal changes in
diabetic individuals [22]. After 30 minutes of
honey mouth rinse there was significant increase
in salivary pH of diabetic participants the same
results were observed atwa et., al. who proved
that honey was effective in preventing plaque
and gingivitis [23]. When the findings were
observed for vinegar mouth rinse the results
identified were parallel to multiple studies in
which its antibacterial, antifungal activities are
compared and are attributed to its ability of
change in salivary pH, the role of vinegar is also
highlighted as a potent irrigant for canal while
Root canal treatment [24,25]. When honey mouth
rinse was compared with vinegar rinse we found
better results with honey which may be due to its
neutralizing properties, however, as vinegar itself
lies in category of week acids it did not show
equality in maintaining the salivary pH of diabetic
individuals.
5. CONCLUSION
Honey and vinegar mouth rinses proved to be
effective natural products in maintaining the pH
of saliva of diabetic individuals.
6. LIMITATIONS
After rinse pH was evaluated after 30 minutes
which is minimum time to strengthen the study.
We recommend recording the pH maximum after
two hours of rinsing.
CONSENT
Each participant was briefed regarding the study
protocol and after that written informed consent
was taken.
ETHICAL APPROVAL
Study was ethically approved by ERC of Baqai
Medical University Karachi.
COMPETING INTERESTS
Authors have declared that no competing
interests exist.
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