D . Ek a Pa el e al. A Randomized Clinical T ial Compa ing he An ibac e ial P ope ies o 0.5% Gua a Lea Ex ac
Mou hwash Wi h 0.1% Tu me ic Ex ac Mou hwash in High-Ca ies-Risk Si ua ions. In . J Med. Pha m. Res., 6 (6):
214‐217, 2025
214
In e na ional Jou nal o Medical
and Pha maceu ical Resea ch
Online ISSN-2958-3683 | P in ISSN-2958-3675
F equency: Bi-Mon hly
A ailable online on: h ps://ijmp .in/
O iginal A icle
A Randomized Clinical T ial Compa ing he An ibac e ial P ope ies o
0.5% Gua a Lea Ex ac Mou hwash Wi h 0.1% Tu me ic Ex ac
Mou hwash in High-Ca ies-Risk Si ua ions
D . Ek a Pa el1, D . Munni Chaudha y2, D . Rahul Pa el3, D . Anagha Ag awal4, D . Pulki Kalyan5, D . Ri u Randad6
1, 6 Assis an P o esso , Depa men o Public Heal h Den is y, Na sinhbhai Pa el Den al College and Hospi al, Sankalchand Pa el
Uni e si y, Visnaga
2 MDS S uden , Depa men o Public Heal h Den is y, Na sinhbhai Pa el Den al College and Hospi al, Sankalchand Pa el
Uni e si y, Visnaga
3 P o esso and Head, Depa men o Public Heal h Den is y, Na sinhbhai Pa el Den al College and Hospi al, Sankalchand Pa el
Uni e si y, Visnaga
4 Associa e P o esso , Depa men o Public Heal h Den is y, Na sinhbhai Pa el Den al College and Hospi al, Sankalchand Pa el
Uni e si y, Visnaga
5 P o esso , Depa men o Public Heal h Den is y, Na sinhbhai Pa el Den al College and Hospi al, Sankalchand Pa el Uni e si y,
Visnaga
A B S T R A C T
Co esponding Au ho :
D . Ek a Pa el
Assis an P o esso , Depa men
o Public Heal h Den is y,
Na sinhbhai Pa el Den al College
and Hospi al, Sankalchand Pa el
Uni e si y, Visnaga .
Recei ed: 14-10-2025
Accep ed: 29-10-2025
A ailable online: 12-11-2025
Goal: To e alua e and con as he an ibac e ial ac i i y o mou hwashes con aining
0.5% gua a lea ex ac and 0.1% u me ic ex ac on S ep ococcus mu ans coun s
in pa ien s who a e a high isk o den al ca i ies.
Me hods: This andomized clinical ial was conduc ed among 60 high-ca ies- isk
pa ien s (30 pe g oup). G oup A ecei ed 0.5% gua a lea es ex ac mou hwash,
while G oup B ecei ed 0.1% u me ic ex ac mou hwash. Pa icipan s insed wi h
10 ml o mou hwash wice daily o 14 days. Sali a samples we e collec ed a
baseline and pos -in e en ion o quan i y S. mu ans colonies using Mi is Sali a ius
Aga . Sali a y pH was eco ded, and pa ien sa is ac ion was assessed using a
s uc u ed ques ionnai e.
Resul s: The gua a mou hwash g oup showed a signi ican ly g ea e educ ion in
S. mu ans coun s (mean educ ion: 2.1 × 10⁵ CFU/ml) compa ed o he u me ic
g oup (mean educ ion: 1.2 × 10⁵ CFU/ml) (p < 0.05). Sali a y pH imp o ed
owa ds alkalini y in he gua a g oup ( om 6.4 o 7.0) compa ed o he u me ic
g oup ( om 6.5 o 6.8). Pa ien sa is ac ion sco es we e highe o gua a (83.3%
a ed as e as pleasan ) han u me ic (63.3%).
Conclusion: Wi hin he pa ame e s o his s udy, 0.5% gua a lea es ex ac
mou hwash was mo e e ec i e as an an ibac e ial agen agains S ep ococcus
mu ans han 0.1% u me ic ex ac mou hwash. I p esen s a p omising na u al
al e na i e o managing den al ca ies isk.
Copy igh © In e na ional Jou nal o
Medical and Pha maceu ical Resea ch
Keywo ds: Gua a lea ex ac , Tu me ic ex ac , Mou hwash, S ep ococcus
mu ans, Den al ca ies.
INTRODUCTION
Den al ca ies emains a p e alen ch onic disease wo ldwide, pa icula ly among high-ca ies- isk popula ions.1
S ep ococcus mu ans is a p ima y e iological agen in he de elopmen o den al ca ies, con ibu ing o enamel
demine aliza ion h ough acid p oduc ion.1 While con en ional an imic obial agen s like chlo hexidine ha e demons a ed
e icacy in educing S. mu ans coun s, hei p olonged use is o en limi ed due o side e ec s such as al e ed as e pe cep ion,
mucosal i i a ion, and oo h s aining. 2 This has p omp ed in e es in explo ing he bal al e na i es wi h an imic obial
p ope ies.3,4
D . Ek a Pa el e al. A Randomized Clinical T ial Compa ing he An ibac e ial P ope ies o 0.5% Gua a Lea Ex ac
Mou hwash Wi h 0.1% Tu me ic Ex ac Mou hwash in High-Ca ies-Risk Si ua ions. In . J Med. Pha m. Res., 6 (6):
214‐217, 2025
215
Gua a (Psidium guaja a) lea es ha e been ecognized o hei an imic obial p ope ies, a ibu ed o bioac i e compounds
such as la onoids, annins, and phenolic acids.3,5 In i o s udies ha e demons a ed ha gua a lea ex ac s exhibi
signi ican an ibac e ial ac i i y agains S. mu ans, sugges ing hei po en ial as a na u al adjunc in o al hygiene p ac ices.3,5
Fo ins ance, a s udy by Kassem e al. (2022)6 epo ed he po en an ibac e ial e ec o gua a lea es on S. mu ans,
highligh ing hei po en ial as an al e na i e o syn he ic an imic obial agen s.3
Tu me ic (Cu cuma longa), pa icula ly i s ac i e compound cu cumin, has also been s udied o i s an imic obial e ec s.5,7
Cu cumin has demons a ed inhibi o y ac i i y agains S. mu ans by in e e ing wi h bac e ial me abolism and bio ilm
o ma ion.5 Resea ch by Waghma e e al. (2011)8 indica ed ha u me ic mou hwash could e ec i ely educe plaque
o ma ion and gingi i is, compa able o chlo hexidine glucona e.
Few compa a i e clinical in es iga ions ha e e alua ed he e icacy o gua a and u me ic ex ac s when used as o al inses
among indi idual p one o den al ca ies, despi e he indi idual da a suppo ing each. 3, 5, 6 By e alua ing and con as ing
he an ibac e ial e ec s o mou hwashes con aining 0.5% gua a lea ex ac and 0.1% u me ic ex ac on S. mu ans coun s
and sali a y pH in high-ca ies h ea indi iduali ies, his s udy seeks o close his gap.
MATERIALS AND METHODOLOGY
STUDY DESIGN AND PARTICIPANT SELECTION:
➢ This s udy was a andomized con olled ial conduc ed in he Na sinhbhai Pa el Den al College and Hospi al's
Depa men o Public Heal h Den is y. E hical app o al was g an ed by he ins i u ional e hics commi ee, and w i en
in o med consen was secu ed om all subjec s.
ELIGIBILITY CRITERIA:
✓ Age 18–45 yea s
✓ High ca ies isk (DMFT ≥ 5, equen suga in ake, poo o al hygiene index > 2.5)
✓ No an ibio ic o an imic obial mou hwash use in he pas 4 weeks
NON- ELIGIBILITY CRITERIA:
• P esence o sys emic illness
• Use o o hodon ic appliances
• A his o y o in ole ance o he bal p oduc s
GETTING MOUTHWASH READY
EXTRACT FROM GUAVA LEAVES (0.5%):
1. F eshly picked gua a lea es we e ca e ully cleaned and allowed o ai -d y in he shade a ambien empe a u e o 7–10
days.
2. D ied lea es we e powde ed using a mechanical g inde .
3. The powde ed lea es we e ex ac ed using 95% e hanol in a Soxhle appa a us o 6–8 hou s.
4. The ex ac was condensed unde acuum condi ions wi h o a y e apo a o o yield a semi-solid mass.
5. The ob ained esidue was econs i u ed in dis illed wa e o o m a 0.5% weigh / olume solu ion.
6. The p epa ed solu ion was passed h ough a 0.45 μm memb ane il e o cla i ica ion and kep in ambe - in ed bo les
a 4°C un il equi ed.
TURMERIC EXTRACT (0.1%):
1. Tu me ic hizomes we e ho oughly cleaned, d ied in shade, and g ound in o a ine powde .
2. E hanolic ex ac ion o he powde ed hizomes was ca ied ou in Soxhle appa a us o six hou s.
3. The ex ac was condensed and adjus ed o achie e a cu cuminoids concen a ion o 0.1%.
4. The esul ing solu ion was il e ed using a 0.45 μm memb ane and p ese ed in ambe -colou ed con aine s a 4°C.
INTERVENTION
➢ To minimize bias and ensu e he eliabili y o he s udy ou comes, a double-blind andomized design was implemen ed.
The p ocedu e was as ollows:
1. Randomiza ion:
➢ A o al o six y eligible pa icipan s we e andomly di ided in o wo equal g oups (n = 30 each) h ough a compu e -
gene a ed andomiza ion sequence.
➢ Pa icipan s in G oup A p o ided wi h 0.5% gua a lea ex ac o al inse, whe eas G oup B was gi en 0.1% u me ic
ex ac o mula ion.
2. Alloca ion Concealmen :
D . Ek a Pa el e al. A Randomized Clinical T ial Compa ing he An ibac e ial P ope ies o 0.5% Gua a Lea Ex ac
Mou hwash Wi h 0.1% Tu me ic Ex ac Mou hwash in High-Ca ies-Risk Si ua ions. In . J Med. Pha m. Res., 6 (6):
214‐217, 2025
216
➢ To ensu e p ope alloca ion concealmen , g oup codes we e sealed in consecu i ely numbe s opaque en elopes,
p epa ed by an independen esea che unin ol ed in pa icipan selec ion o da a analysis. The en elopes we e opened
solely a he ime o dis ibu ing he o al inses.
3. Blinding o Pa icipan s:
➢ Pa icipan s we e blinded since he mou hwashes had iden ical colo s, consis ency, and packaging.
➢ To ensu e pa icipan blinding, pa icipan s we e no old which mou hwash hey we e ge ing.
➢ Two g oups we e andomly selec ed om among he pa icipan s. Mou hwash con aining 0.5% gua a lea ex ac was
gi en o G oup A, while mou hwash con aining 0.1% u me ic ex ac was gi en o G oup B. Fo 14 days, each
pa icipan was old o inse wice a day (mo ning and nigh ) o 60 seconds wi h 10 cc o he designa ed mou hwash.
DATA COLLECTION
MICROBIOLOGICAL ANALYSIS: Uns imula ed sali a y specimens we e ob ained om e e y pa icipan bo h be o e
and a e he in e en ion pe iod. The samples we e se ially dilu ed, inocula ed on o Mi is Sali a ius Aga supplemen ed
wi h Baci acin, ollowed incuba ion a 37°C o 48 hou s. The S. mu ans Colonies we e ecognized and quan i ied as
CFU/ml.
SALIVARY PH MEASUREMENT: The sali a y pH was de e mined u ilizing a pH s ips.
PATIENT ACCEPTENCE: Pa icipan app o al ega ding as e, eshness, and willingness o con inue use was e alua ed
h ough a s uc u ed 5-poin Like scale ques ionnai e.
ANALYTICAL STATISTICS
➢ S a is ical e alua ion was conduc ed using SPSS e sion 20. Pai ed - es s we e applied o compa e p e- and pos -
in e en ion da a wi hin each g oup, whe eas independen - es s examined in e g oup a ia ions. A p- alue o less
han 0.05 was conside ed s a is ically signi ican .
RESULTS
➢ The s udy success ully e alua ed he ou comes o all en olled pa icipan s. The key indings a e summa ized below:
Table 1. Compa ison o S ep ococcus mu ans coun s (CFU/ml) be ween g oup.
PARAMETER
GROUP A
GROUP B
P VALUE
S. mu ans
Baseline
3.8 × 10⁵ ± 0.6 × 10⁵
3.9 × 10⁵ ± 0.5 × 10⁵
0.72
Pos - in e en ion
1.7 × 10⁵ ± 0.4 × 10⁵
2.7 × 10⁵ ± 0.5 × 10⁵
0.001*
Reduc ion
2.1 × 10⁵
1.2 × 10⁵
0.001*
Table 2. Compa ison o Sali a y pH be ween g oups.
PARAMETER
GROUP A
GROUP B
P VALUE
Sali a y pH
Baseline
6.4 ± 0.2
6.5 ± 0.3
0.18
Pos - in e en ion
7.0 ± 0.3
6.8 ± 0.3
0.02*
Table 3. Pa ien sa is ac ion ega ding mou hwash as e and accep abili y
PARAMETER
GROUP A
GROUP B
P VALUE
Pleasan Tas e
83.3%
63.3%
0.03*
Willingness o Con inue
use
80%
60%
0.04*
ANTIBACTERIAL EFFECT:
➢ The g oup using he 0.5% gua a lea es ex ac mou hwash showed a s a is ically signi ican g ea e educ ion in S.
mu ans om mean 3.8 × 10⁵ o 1.7 × 10⁵ CFU/ml compa ed o he g oup using he 0.1% u me ic ex ac mou hwash
educed om mean 3.9 × 10⁵ o 2.7 × 10⁵ CFU/ml. The educ ion was signi ican ly highe in he gua a g oup (p =
0.001).
SALIVARY PH: A mo e signi ican inc ease owa ds alkaline pH was obse ed in he gua a ex ac g oup.
PATIENT SATISFACTION: The ques ionnai e esul s indica ed Gua a g oup: 83.3% a ed as e pleasan , 80% willing
o con inue use. While Tu me ic g oup: 63.3% a ed as e pleasan , 60% willing o con inue use.
D . Ek a Pa el e al. A Randomized Clinical T ial Compa ing he An ibac e ial P ope ies o 0.5% Gua a Lea Ex ac
Mou hwash Wi h 0.1% Tu me ic Ex ac Mou hwash in High-Ca ies-Risk Si ua ions. In . J Med. Pha m. Res., 6 (6):
214‐217, 2025
217
DISCUSSION
The indings o his s udy align wi h exis ing li e a u e suppo ing he an imic obial p ope ies o Psidium guaja a. The
enhanced e ec i eness o gua a lea ex ac may esul om i s abundan la onoid and annin con en , compounds
ecognized o supp essing bac e ial p oli e a ion and a achmen . Al hough u me ic exhibi s an imic obial po en ial, he
0.1% o mula ion u ilized in his in es iga ion p o ed o be less e icacious han he 0.5% gua a ex ac .
Findings indica e ha 0.5% gua a lea es ex ac is a iable and po en na u al al e na i e o educing he bac e ial load
associa ed wi h den al ca ies. I s e icacy, coupled wi h good pa ien accep abili y, makes i a s ong candida e o u he
de elopmen as a comme cial mou hwash.
LIMITATIONS
➢ Small sample size, sho s udy du a ion (14 days), and absence o a posi i e con ol (e.g., chlo hexidine) limi
gene alizabili y.
RECOMMENDATIONS
➢ Longe ollow-up, la ge coho s, and inclusion o a s anda d compa a o will s eng hen he e idence o gua a as a
na u al mou hwash.
CONCLUSION
➢ The 0.5% gua a lea es ex ac mou hwash p o ed mo e e ec i e han 0.1% u me ic ex ac mou hwash in educing
S. mu ans coun s, imp o ing sali a y pH, and achie ing highe pa ien accep abili y in high-ca ies- isk pa ien s. Gua a
mou hwash p esen s a p omising, a o dable, and sa e he bal al e na i e o ca ies inhibi ion.
ACKNOWLEDGEMENTS:
The au ho s would like o hank he S uden S a -up & Inno a ion Policy (SSIP 2.0) o i s inancial assis ance. Educa ion
Depa men , Go e nmen o Guja a , h ough he SSIP Cell a Sankalchand Pa el Uni e si y (P ojec ID:
SPU_SSIP2.0_47). We also ex end ou g a i ude o he pa icipan s o his s udy.
REFERENCES
1. Fea he s one, J. D. (2008). Den al ca ies: A dynamic disease p ocess. Aus alian Den al Jou nal, 53(3), 286–291.
h ps://doi.o g/10.1111/j.1834-7819.2008.00064.x
2. Löe, H., & Schiø , C. R. (1970). The e ec o mou h inses and opical applica ion o chlo hexidine on he de elopmen
o den al plaque and gingi i is in man. Jou nal o Pe iodon al Resea ch, 5(2), 79–83. h ps://doi.o g/10.1111/j.1600-
0765.1970. b00696.x
3. P abu, G. R., Gnanamani, A., & Sadulla, S. (2006). Guaija e in—a plan la onoid as po en ial an iplaque agen
agains S ep ococcus mu ans. Jou nal o Applied Mic obiology, 101(2), 487–495. h ps://doi.o g/10.1111/j.1365-
2672.2006.02922.x
4. Dineshkuma , B., Rajakuma , R., & Kuma a el, S. (2010). An imic obial ac i i y o Psidium guaja a agains den al
ca ies causing pa hogens. In e na ional Jou nal o Pha mTech Resea ch, 2(1), 111–114.
5. Ju enka, J. S. (2009). An i-in lamma o y p ope ies o cu cumin, a majo cons i uen o Cu cuma longa: A e iew o
p eclinical and clinical esea ch. Al e na i e Medicine Re iew, 14(2), 141–153.
6. Kassem, E. M. (2022). An ibac e ial e ec o gua a lea es ex ac mou hwash e sus 0.1% u me ic ex ac
mou hwash in high-ca ies- isk pa ien s: A andomized clinical ial. Jou nal o Indian Socie y o Pe iodon ology, 26(4),
373–379. h ps://doi.o g/10.4103/jisp.jisp_362_21
7. Mali, G. V., & Bansode, S. H. (2019). Compa a i e e alua ion o e icacy o he bal mou hwashes con aining Cu cuma
longa and Psidium guaja a on S ep ococcus mu ans le els: A clinical s udy. Jou nal o Indian Socie y o Pedodon ics
and P e en i e Den is y, 37(4), 393–399. h ps://doi.o g/10.4103/JISPPD.JISPPD_242_19
8. Waghma e, P. F., Chaudha i, A. U., Ka hadka , V. M., & Jamkhande, A. S. (2011). Compa a i e e alua ion o u me ic
and chlo hexidine glucona e mou hwash in p e en ion o plaque o ma ion and gingi i is: A clinical and
mic obiological s udy. Jou nal o Con empo a y Den al P ac ice, 12(4), 221–224. h ps://doi.o g/10.5005/jp-jou nals-
10024-1039
