Moch Fu uchul A i in e al. Fo mula ion o D ied C ude Papain in an Ex olia ing Sc ub C eam: Op imiza ion Using
Mini ab 20 Response Op imize and E alua ion o P o eoly ic Ac i i y ia Release Tes . In . J Med. Pha m. Res., 6 (6):
340‐351, 2025
340
In e na ional Jou nal o Medical
and Pha maceu ical Resea ch
Online ISSN-2958-3683 | P in ISSN-2958-3675
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A ailable online on: h ps://ijmp .in/
O iginal A icle
Fo mula ion o D ied C ude Papain in an Ex olia ing Sc ub C eam:
Op imiza ion Using Mini ab 20 Response Op imize and E alua ion o
P o eoly ic Ac i i y ia Release Tes
Moch Fu uchul A i in1, Kosasih Kosasih1, Jessica In an Fe lia1
1 Facul y o Pha macy, Uni e si as Pancasila, Jaka a 12640, Indonesia
A B S T R A C T
Co esponding Au ho :
Moch Fu uchul A i in
Facul y o Pha macy, Uni e si as
Pancasila, Jaka a 12640,
Indonesia.
Recei ed: 17-09-2025
Accep ed: 05-10-2025
A ailable online: 12-11-2025
Papain is a p o eoly ic enzyme known o i s ex olia ing p ope ies, capable o
lysing dead skin cells on he skin’s su ace. This s udy aimed o o mula e and
op imize a sc ub c eam con aining d ied c ude papain as an ac i e ex olian . A 2²
ac o ial design was employed, wi h s ea ic acid (10%)– ie hanolamine (3%–4%)
and ce yl alcohol (2%–2.5%) as o mula ion a iables. The sc ub c eam was
e alua ed o i s physical and chemical cha ac e is ics, including iscosi y,
lowabili y, sp eadabili y, c eam ype, pH, and p o eoly ic ac i i y. P o eoly ic
elease was assessed using a F anz di usion cell. The esul ing o mula ions
exhibi ed iscosi ies anging om 60,000 o 102,000 cP a 2 pm, sp eadabili y
be ween 76.74 and 116.12 g•cm/sec, and pH alues om 7.64 o 7.95. P o eoly ic
ac i i y anged om 1.02 o 4.24 TU/mg. S ea ic acid– ie hanolamine signi ican ly
in luenced sp eadabili y, p o eoly ic ac i i y, and pH, while ce yl alcohol a ec ed
all measu ed pa ame e s. The op imized o mula ion consis ed o s ea ic acid–
ie hanolamine (10%; 4%) and ce yl alcohol (2%). The elease mechanism o
papain om he sc ub c eam was iden i ied as a combina ion o Fickian di usion
and ma ix elaxa ion. These indings suppo he po en ial o d ied c ude papain as
a iable ac i e ing edien in ex olia ing sc ub c eam o mula ions.
Copy igh © In e na ional Jou nal o
Medical and Pha maceu ical Resea ch
Keywo ds: Papain; sc ub c eam o mula ion; ac o ial design; p o eoly ic ac i i y;
F anz di usion cell; ex olia ing cosmeceu ical.
INTRODUCTION
The accumula ion o dead skin cells on he skin’s su ace con ibu es o a dull complexion. To p e en his, ex olia o s
a e commonly used. Papain, a p o eoly ic enzyme de i ed om he papaya plan (Ca ica papaya L.), is a na u al
ex olian . I wo ks by lysing dead skin cells adhe ed o he skin, esul ing in a smoo he and b igh e appea ance [1,2].
Ka dono e al. epo ed ha a 1% papain lo ion can b igh en he skin by inhibi ing he y osinase enzyme [3], while o he
s udies ha e shown ha e en 0.2% papain can enhance skin b igh ness h ough ex olia ion [4].
Angg aini e al. o mula ed papaya gum in o a wa e -in-oil (W/O) emulsion c eam [5]. Howe e , W/O c eams a e
gene ally less com o able o opical use due o hei g easy ex u e and di icul y in emo al [6]. To imp o e use
com o —ensu ing ease o applica ion, non-s ickiness, a cooling sensa ion, non-comedogenici y, and ease o emo al—
papain coa se powde was o mula ed in o an oil-in-wa e (O/W) c eam base. This app oach also enhances he s abili y
and ex olia ing e icacy o papain [7].
Ex olia ing sc ubs a e designed o ac h ough bo h enzyma ic and mechanical mechanisms, wi h he addi ion o
sc ubbing agen s o acili a e he emo al o dead skin cells. To maximize enzyma ic ex olia ion, 10% c ude papain
Moch Fu uchul A i in e al. Fo mula ion o D ied C ude Papain in an Ex olia ing Sc ub C eam: Op imiza ion Using
Mini ab 20 Response Op imize and E alua ion o P o eoly ic Ac i i y ia Release Tes . In . J Med. Pha m. Res., 6 (6):
340‐351, 2025
341
powde was inco po a ed in o he o mula ion [6,8-10]. T ie hanolamine-s ea a e was used as an in si u emulsi ie o
imp o e he s abili y o he O/W emulsion.
This s udy employed a 2² ac o ial design wi h wo o mula ion ac o s: s ea ic acid– ie hanolamine as he emulsi ie and
ce yl alcohol as he hickene .[11] The concen a ion anges used we e s ea ic acid– ie hanolamine (10%; 3–4%) and
ce yl alcohol (2–2.5%). The ex olia ing sc ub c eam was e alua ed o o ganolep ic p ope ies, homogenei y, c eam ype,
iscosi y and low beha io , sp eadabili y, pH, and p o eoly ic ac i i y. P o eoly ic elease was assessed using a F anz
di usion cell. To de e mine he e ec s o s ea ic acid– ie hanolamine and ce yl alcohol on he o mula ion
cha ac e is ics, ac o e ec and in e ac ion analyses we e conduc ed using Mini ab 20 so wa e.[12] The op imal
o mula ion was iden i ied using he Mini ab 20 Response Op imize [6,13]
MATERIALS AND METHODS
2.1. Ma e ials
C ude papain was om Nanning Pangbo Biological Enginee ing Co., L d., China. Ty osine and casein we e pu chased
om Sigma-Ald ich, Ge many, while cys eine HCl monohyd a e and S-pack di usion memb anes we e om Me ck,
Ge many. The ins umen s used included a B ook ield iscome e (RV ype), USA; a UV-Vis spec opho ome e
(Shimadzu UV-1800), Japan; and a F anz di usion cell (Logan Ins umen s), USA.
2.2. Me hods
2.2.1. P epa a ion o Casein Subs a e
One g am o casein was dispe sed in 50 mL o 0.05 M sodium phospha e bu e and hea ed in a wa e ba h a 40 °C o
30 min. A e cooling o oom empe a u e, he pH was adjus ed o 6.0 ± 0.1 using 0.05 M ci ic acid. The casein solu ion
was dilu ed wi h pu i ied wa e o make a inal olume o 100 mL (designa ed as Solu ion A). [14]
2.2.2. P epa a ion o Phospha e–Cys eine–Ede a e Bu e
The p epa a ion o phospha e–cys eine–ede a e bu e in ol ed dissol ing 3.55 g o anhyd ous sodium phospha e in 400
mL o pu i ied wa e , adding 7 g o sodium ede a e, and 3.05 g o cys eine HCl monohyd a e. The bu e was mixed and
he pH was adjus ed o 6.0 ± 0.1 using 1 N HCl o 1 N NaOH. The inal olume was 500 mL wi h pu i ied wa e
(designa ed as Solu ion B). [15]
2.2.3. P epa a ion o C ude Papain Solu ion
Exac ly 100.00 mg o c ude papain powde was weighed and dissol ed in Solu ion B o a inal olume o 100 mL. F om
his s ock solu ion, 2 mL was pipe ed in o a 50 mL olume ic lask and dilu ed wi h Solu ion B o ob ain 50 mL o
wo king solu ion (designa ed as Solu ion C). [16][17][18]
2.2.4. P epa a ion o Ty osine S anda d Cu e
Ty osine (10.00 mg) was dissol ed in pu i ied wa e and dilu ed o a inal olume o 100 mL, yielding Solu ion A. A
se ies o wo king solu ions, con aining 25, 35, 45, 55, 65, 75, and 85 ppm y osine, was p epa ed om he s ock solu ion.
Abso bance was measu ed a he maximum wa eleng h using a UV-Vis spec opho ome e . A calib a ion cu e was
cons uc ed by plo ing abso bance agains concen a ion, yielding he eg ession equa ion: [19][20][21]
Y = −0.0059 + 0.0092X, wi h a co ela ion coe icien R² = 0.9985.
2.2.5. Measu emen o P o eoly ic Ac i i y (AOAC Me hod)
P o eoly ic ac i i y was measu ed using he AOAC me hod. One g am o casein was dispe sed in 50 mL o 0.05 M
sodium phospha e bu e and hea ed in a wa e ba h a 40 °C o 30 min. A e cooling, he pH was adjus ed o 6.0 ± 0.1
using 0.05 M ci ic acid. The solu ion was hen dilu ed wi h pu i ied wa e o a inal olume o 100 mL, designa ed as
Solu ion A. Phospha e–cys eine–ede a e bu e (Solu ion B) was p epa ed by dissol ing 3.55 g o anhyd ous sodium
phospha e in 400 mL o pu i ied wa e , ollowed by he addi ion o 7 g o sodium ede a e and 3.05 g o cys eine HCl
monohyd a e. Solu ion B was mixed, and he pH was adjus ed o 6.0 ± 0.1 using 1 N HCl o 1 N NaOH. The inal olume
was 500 mL, adjus ed wi h pu i ied wa e . App oxima ely 100 mg o d ied c ude papain was dissol ed in 100 mL o
Solu ion B. F om his s ock, 2 mL was pipe ed in o a 50 mL olume ic lask and dilu ed wi h Solu ion B o ob ain 50
mL o wo king solu ion (designa ed as Solu ion C). Two es ubes we e p epa ed and designa ed as he sample (S) and
he blank (B). In he sample ube, 5 mL o Solu ion A was added and p eincuba ed a 40 °C o 10 min. Then, 1 mL o
Solu ion C and 1 mL o Solu ion B we e mixed and incuba ed a 40 °C o an hou . Th ee mL o TCA solu ion was added,
and he mix u e was o exed ho oughly. Bo h ubes we e hen incuba ed a 40 °C o an addi ional 30 min and il e ed
using Wha man No. 42 il e pape . The il a e was collec ed, and abso bance was measu ed a he maximum wa eleng h
using a UV-Vis spec opho ome e . [22]
Moch Fu uchul A i in e al. Fo mula ion o D ied C ude Papain in an Ex olia ing Sc ub C eam: Op imiza ion Using
Mini ab 20 Response Op imize and E alua ion o P o eoly ic Ac i i y ia Release Tes . In . J Med. Pha m. Res., 6 (6):
340‐351, 2025
342
Papain ac i i y was de e mined using he y osine s anda d cu e o i s linea eg ession equa ion:
Y = -0.0059 + 0.0092X, R2 = 0.9985
The p o eoly ic ac i i y (A, in TU/mg) was de e mined using he ollowing o mula:
Cx 100
Wx50
2x10
1xA
Whe e: (C): concen a ion o y osine (mg/mL), (W): weigh o d ied c ude papain (mg), and (A): p o eoly ic ac i i y
(TU/mg)
2.2.6. Fo mula ion o D y C ude Papain Sc ub C eam
The oil and aqueous phases o he c eam base we e hea ed sepa a ely in a wa e ba h a 70–75 °C. Me hyl pa aben and
p opyl pa aben we e added and mixed in p opylene glycol. The oil phase was g adually added o he aqueous phase
d opwise unde con inuous s i ing. Homogeniza ion was done a he op imal speed and du a ion un il a s able c eam
base o med. [23][24]
2.2.7. De e mina ion o C eam Type
A o al o 0.1 g o sc ub c eam was dispe sed and mixed in 10 mL o pu i ied wa e . A e ho ough mixing, he sample
was examined unde a mic oscope. One d op o he esul ing mix u e was placed on a mic oscope slide, ollowed by he
addi ion o me hylene blue solu ion. The p esence o a uni o m blue colo a ion in he ex e nal phase indica ed an oil-in-
wa e (O/W) emulsion ype. [24][25]
2.2.8. Viscosi y and Flow Beha io
Viscosi y was measu ed using a B ook ield iscome e (RVL ype) equipped wi h an app op ia e spindle. Measu emen s
we e conduc ed ac oss a ange o o a ional speeds ( pm), p og essing om he lowes o he highes se ing. Each
eading was eco ded a 10-minu e in e als, ensu ing scale alues exceeded 10 uni s o accu acy. Viscosi y, exp essed
in cen ipoise (cP), was calcula ed using he ollowing equa ion:
Viscosi y = scale x mul iplica ion ac o (cP) (2)
Shea o ce (F) = scale x K (dyne/cm2) (3)
K s = 7187.00 dyne/cm²
The e alua ion o low beha io was conduc ed by cons uc ing a heog am, plo ing shea o ce (F) on he x-axis agains
shea a e ( pm) on he y-axis using g aph pape . [26][27][28]
2.2.9. Sp eadabili y Tes
Sp eadabili y was e alua ed by placing 0.5 g o sc ub c eam on o a wa ch glass o mic oscope slide posi ioned o e
g aph pape . A pe i dish was placed a op he sample, and weigh s o 50, 100, and 200 g we e applied sequen ially o 1
minu e each. The diame e s o he sp ead a ea we e measu ed in mul iple di ec ions, and he mean alue was eco ded.
[29]
2.2.10. pH Measu emen
Be o e measu emen , he pH elec ode was calib a ed using s anda d bu e solu ions. A 10 g o sc ub c eam was
dissol ed in 100 mL o pu i ied wa e , and he pH was de e mined using a calib a ed pH me e . The accep able pH ange
o he o mula ion was 3.5–8.0. [30]
2.2.11. Papain Release Tes Using F anz Di usion Cell
One g am o sc ub c eam was applied o an S-Pak memb ane, which was hen moun ed in a F anz di usion cell. The
memb ane was secu ed using a clamp ing o p e en ai ing ess. Phospha e bu e (5 mL, pH 7.4) was added o bo h he
dono and ecep o compa men s. A magne ic s i ba was placed in he ecep o chambe , and he sys em was sealed and
main ained a 37 °C. A e 30 minu es, a 5 mL aliquo was collec ed om he ecep o compa men o papain elease.
[31][32]
2.2.12. Fac o ial Design Analysis
Expe imen al da a, including iscosi y, sp eadabili y, pH, and p o eoly ic ac i i y, we e analyzed using Mini ab 20
so wa e o e alua e he e ec s o o mula ion ac o s and hei in e ac ions. (Table 1) The op imum o mula ion was
de e mined using a esponse op imize . The op imiza ion c i e ia included minimizing iscosi y, maximizing
sp eadabili y and papain ac i i y, and main aining he pH wi hin he ange o 3.5–8.0. [33][34]
Table 1. Fo mula o d ied c ude papain sc ub c eam wi h ac o ial design 22
Moch Fu uchul A i in e al. Fo mula ion o D ied C ude Papain in an Ex olia ing Sc ub C eam: Op imiza ion Using
Mini ab 20 Response Op imize and E alua ion o P o eoly ic Ac i i y ia Release Tes . In . J Med. Pha m. Res., 6 (6):
340‐351, 2025
343
Ing edien
Concen a ion (%)
F1
F2
F3
F4
D ied c ude papain
10
10
10
10
S ea ic acid
10
10
10
10
T ie hanolamine
3
4
3
4
Se yl alcohol
2
2
2.5
2.5
P opylene glycol
10
10
10
10
P opyl pa aben
0.05
0.05
0.05
0.05
Me hyl pa aben
0.15
0.15
0.15
0.15
Polye hylene
5
5
5
5
Aquades ad
100
100
100
100
2.2.13. Op imum Sc ub C eam Fo mula ion
Based on he op imiza ion esul s ob ained using Mini ab 20, he ideal o mula ion—comp ising 10% s ea ic acid and 4%
ie hanolamine—co esponded o Fo mula 2. The op imized o mula ion was subsequen ly employed o e alua e he
elease p o ile o papain. [35]
2.2.14. Papain Release Analysis om Sc ub C eam
Papain elease om he op imized sc ub c eam (Fo mula 2) was e alua ed using a F anz di usion cell, a alida ed
me hod o assessing enzyme di usion om semi-solid ma ices. [23][31] Samples we e wi hd awn om he ecep o
compa men a p ede e mined in e als (15–180 min), and papain ac i i y was quan i ied ia UV-Vis spec opho ome y,
exp essed in TU/mg. The cumula i e elease p o ile is p esen ed in Table 2. [36]
RESULTS
Table 2. Cumula i e papain elease ac i i y agains a ia ions in sampling ime, using a F anz di usion appa a us
Time (minu es)
15
30
45
60
90
120
150
180
Cumula i e papain
ac i i y (M )
3.58
5.76
7.15
8.38
9.60
10.65
11.22
12.41
Table 3. Comp ehensi e measu emen esul s o d ied c ude papain sc ub c eam
Fo mula ion
C eam
ype
Flow
p ope ies
Viscosi y
(a pm 2, cP )
Sp eadabili y
(g.cm/sec)
Ph
P o eoly ic ac i i y
(TU/mg)
F1
O/W
Plas ic
86,000±800
115.37±0.63
7.64±0.06
3.41±0.06
F2
O/W
Plas ic
60,000±769
116.12±0.53
7.85±0.04
4.24±0.01
F3
O/W
Plas ic
102,000±900
76.74±2.59
7.91±0.04
1.02±0.08
F4
O/W
Plas ic
92,000±840
114.97±2.36
7.95±0.03
1.42±0.06
Table 4. Analysis esul s o ac o e ec s and hei in e ac ions on iscosi y esponse
Fac o
E ec
Signi icance (p- alue)
TEA-s ea ic
-20.667
0.266
Ce yl alcohol
45.333
0.031
TEA-s ea ic*Ce yl alcohol
-6.667
0.710
Moch Fu uchul A i in e al. Fo mula ion o D ied C ude Papain in an Ex olia ing Sc ub C eam: Op imiza ion Using
Mini ab 20 Response Op imize and E alua ion o P o eoly ic Ac i i y ia Release Tes . In . J Med. Pha m. Res., 6 (6):
340‐351, 2025
344
Figu e 1. Plo showing main e ec ac o s (A) and hei in e ac ion (B) on iscosi y ( iskosi as) esponse
Table 5. Analysis esul s o ac o e ec s and hei in e ac ions on he sp eadabili y esponse.
Fac o
E ec
Signi icance (p- alue)
TEA-s ea ic
18.735
0,000
Ce yl alcohol
-19.492
0,000
TEA-s ea ic*Ce yl alcohol
19.888
0,000
Figu e 2. Plo showing main e ec (A) ac o s and hei in e ac ion (B) on he sp eadabili y (daya seba ) esponse.
Table 6. Analysis esul s o ac o e ec s and hei in e ac ions on pH alues
Fac o
E ec
Signi icance (p- alue)
TEA-s ea ic
0.1250
0.001
Ce yl alcohol
0.1850
0.000
TEA-s ea ic*Ce yl alcohol
-0.0917
0.007
Moch Fu uchul A i in e al. Fo mula ion o D ied C ude Papain in an Ex olia ing Sc ub C eam: Op imiza ion Using
Mini ab 20 Response Op imize and E alua ion o P o eoly ic Ac i i y ia Release Tes . In . J Med. Pha m. Res., 6 (6):
340‐351, 2025
345
A B
Figu e 3. Plo showing main e ec ac o s (A, le )) and hei in e ac ion (B, igh ) on pH esponse.
Table 7. Analysis esul s o ac o e ec s and hei in e ac ions on Papain ac i i y
A B
Figu e 4. Plo showing main e ec ac o s (A) and hei in e ac ion (B) on p o eoly ic ac i i y (ak i i as
p o eoli ik).
Fac o
E ec
Signi icance (p- alue)
TEA-s ea ic
0.6140
0.000
Ce yl alcohol
-2.6037
0.000
TEA-s ea ic* Ce yl alcohol
-0.2114
0.000
Moch Fu uchul A i in e al. Fo mula ion o D ied C ude Papain in an Ex olia ing Sc ub C eam: Op imiza ion Using
Mini ab 20 Response Op imize and E alua ion o P o eoly ic Ac i i y ia Release Tes . In . J Med. Pha m. Res., 6 (6):
340‐351, 2025
346
*) ie ano ( ie anolamine); se il al (ce yl alcohol); Ta g ( a ge ); ak i i a (ac i i y); iskosi ( iscosi y); daya seba
(sp aedabili y)
Figu e 5. Analysis o de e mining he op imal o mula using Mini ab 20's esponse op imize .
Tabel 8. Model, equa ion and cons an alues o papain elease mechanism model
Model
Equa ion
R2 & n
Ze o-o de
M = 4.4623+0.0429
0.9118
Fi s -o de
lnM = ln1.5528 + 0.0032
0.7080
Higuchi
M = 0.9519 + 0.8745 √
0.9754
Ko smeye -Peppas
LnM / M∾ = Ln 0.0159+0.4768 Ln
0.9745 & 0.4768
Figu e 6. Mechanism o papain elease om sc ap c eam: A, ze o-o de ; B, i s -o de ; C, Higuchi, and D,
Ko smeye -Peppas me hod.
DISCUSSION
P o eoly ic ac i i y was e alua ed using F anz di usion cells by measu ing he enzyma ic hyd olysis o casein subs a es
by papain, which eleases y osine as a quan i iable p oduc . The p o eoly ic ac i i y alues o o mula ions F1–F4
anged om 1.02 o 4.24 TU/mg, indica ing e ec i e enzyma ic unc ion wi hin he c eam ma ices (Table 2). [31]
Table 3 p esen s he comp ehensi e physicochemical cha ac e is ics o he d ied c ude papain sc ub c eam o mula ions.
All o mula ions (F1–F4) exhibi ed a mul iple emulsion ype (M/A) and demons a ed plas ic low beha io . This
heological p ope y is ypical o suspension and emulsion sys ems and is a ibu ed o loccula ion. [22] Plas ic low is
cha ac e ized by a yield alue, below which he o mula ion beha es elas ically. This allows he c eam o emain adhe ed
o he skin su ace un il i is physically emo ed, enhancing i s opical e en ion. Viscosi y measu emen s a 2 pm
e ealed alues o 86,000, 60,000, 102,000, and 92,000 cP o F1 h ough F4, espec i ely, con i ming he semisolid
consis ency sui able o de mal applica ion. [38] Sp eadabili y, which e lec s he ease o applica ion [37] and uni o m
dis ibu ion on he skin, showed mean alues o 115.37 ± 0.63, 116.12 ± 0.53, 76.74 ± 2.59, and 114.97 ± 2.36 mm o
F1–F4, espec i ely. The pH alues o he o mula ions anged om 7.64 o 7.95, aligning wi h he op imal pH ange o
Moch Fu uchul A i in e al. Fo mula ion o D ied C ude Papain in an Ex olia ing Sc ub C eam: Op imiza ion Using
Mini ab 20 Response Op imize and E alua ion o P o eoly ic Ac i i y ia Release Tes . In . J Med. Pha m. Res., 6 (6):
340‐351, 2025
347
papain ac i i y ( ypically pH 6.0–8.0). [34] The sligh ly alkaline pH may also be a ibu ed o he p esence o
ie hanolamine, a commonly used emulsi ie and pH adjus e wi h basic p ope ies.
The Impac o Di e en Fac o s and Thei In e ac ions on Viscosi y
Table 4 and Figu e 1 summa ize he e ec s o indi idual o mula ion componen s and hei in e ac ions on he iscosi y
esponse o sc ub c eam o mula ions. S a is ical analysis e ealed ha ce yl alcohol had a signi ican posi i e e ec on
iscosi y (e ec = 45.333; p = 0.031), while TEA-s ea ic acid showed a non-signi ican nega i e e ec . The in e ac ion
be ween TEA-s ea ic acid and ce yl alcohol was also non-signi ican . Figu e 1 illus a es he main e ec s (A) and
in e ac ion e ec s (B) o hese ac o s on iscosi y. The plo clea ly shows ha inc easing he concen a ion o ce yl
alcohol leads o a ma ked ise in iscosi y. This inc ease is a ibu ed o ce yl alcohol’s ole as a consis ency enhance . As
a a y alcohol, ce yl alcohol in e ac s wi h bo h he aqueous and oil phases, o ming s uc u ed ne wo ks ha ap wa e
and inc ease he in e nal esis ance o he emulsion. This beha io is pa icula ly p onounced in oil-in-wa e (O/W)
sys ems, whe e ce yl alcohol con ibu es o emulsion s abili y and hickening by educing in e acial ension and
p omo ing gel-like consis ency. S udies ha e shown ha ce yl alcohol enhances he iscosi y and ex u e o cosme ic
emulsions by o ming lamella s uc u es and inc easing wa e e en ion wi hin he ma ix. [27][39][40]
The Impac o Di e en Fac o s and Thei In e ac ions on Sp eadabili y
The sp eadabili y o he sc ub c eam o mula ions was signi ican ly in luenced by bo h indi idual ac o s and hei
in e ac ion, as shown in he ac o ial design analysis. The in e ac ion plo be ween TEA-s ea a e and ce yl alcohol
e ealed ha inc easing he concen a ion o TEA-s ea a e emulsi ie a 2.5% ce yl alcohol led o a ma ked imp o emen
in sp eadabili y (Table 5 and Figu e 2). This enhancemen is a ibu ed o a co esponding dec ease in iscosi y, which
acili a es easie applica ion and dis ibu ion o he c eam on he skin su ace. In con as , when ce yl alcohol
concen a ion was educed o 2.0%, he sp eadabili y dec eased. This beha io is consis en wi h he ole o ce yl alcohol
as a consis ency enhance ; lowe le els esul in less s uc u al in eg i y and educed wa e -binding capaci y, he eby
inc easing iscosi y and limi ing he c eam’s abili y o sp ead uni o mly. These indings align wi h p e ious s udies
demons a ing ha emulsi ie concen a ion and a y alcohol con en di ec ly a ec he heological and senso y
p ope ies o oil-in-wa e (O/W) emulsions. TEA-s ea a e con ibu es o emulsi ica ion and s abili y, while ce yl alcohol
enhances ex u e and modula es iscosi y, bo h o which a e c i ical o achie ing op imal sp eadabili y in opical
o mula ions [27][34][41].
The Impac o Di e en Fac o s and Thei In e ac ions on pH alues.
Figu e 3 illus a es he main e ec s (A) and in e ac ion e ec s (B) o o mula ion a iables on he pH esponse o he
sc ub c eam. As shown in Table 6, bo h he indi idual ac o s—TEA-s ea ic and ce yl alcohol—and hei in e ac ion
signi ican ly in luenced he pH o he o mula ions.
An inc ease in TEA-s ea ic concen a ion led o a co esponding ise in pH. This e ec is a ibu ed o he p esence o
ie hanolamine (TEA), which unc ions as a neu alizing agen . TEA eac s wi h s ea ic acid o o m TEA-s ea a e, a
soap-based emulsi ie ha con ibu es o he alkalini y o he sys em. Consequen ly, highe concen a ions o TEA esul
in ele a ed pH alues in he inal p epa a ion. [27]
The in e ac ion plo in Figu e 3B u he demons a es ha he pH inc ease was mo e p onounced a a ce yl alcohol
concen a ion o 2.5% compa ed o 2.0%. This sugges s ha ce yl alcohol may modula e he bu e ing capaci y o
emulsi ica ion e iciency o he TEA-s ea a e sys em [39][41], he eby in luencing he inal pH. These indings a e
consis en wi h p e ious epo s highligh ing he ole o TEA in pH modula ion and he s abilizing e ec o a y alcohols
in emulsion sys em.
The Impac o Di e en Fac o s and Thei In e ac ions on C ude Papain Ac i i y.
Table 7 p esen s he analysis o a iance esul s, indica ing ha all h ee o mula ion ac o s signi ican ly in luenced he
p o eoly ic ac i i y o he c ude papain sc ub c eam. An inc ease in ce yl alcohol concen a ion led o a educ ion in
p o eoly ic ac i i y. This e ec is a ibu ed o he co esponding inc ease in iscosi y, which impedes he di usion o
papain om he c eam ma ix. This ela ionship aligns wi h he S okes–Eins ein di usion heo y, which s a es ha he
di usion coe icien (D) o a solu e is in e sely p opo ional o he iscosi y (η) o he medium. As iscosi y inc eases,
molecula mobili y dec eases, esul ing in a lowe di usion coe icien and, consequen ly, a slowe elease a e o he
ac i e compound. As illus a ed in Figu e 4A, inc easing he concen a ion o he TEA-s ea ic emulsi ie signi ican ly
enhanced he s abili y o he papain enzyme, leading o a no able inc ease in p o eoly ic ac i i y. This may be due o
imp o ed emulsion s abili y and mic oen i onmen al p o ec ion p o ided by he emulsi ie , which helps p ese e
enzyma ic unc ionali y du ing o mula ion and applica ion. [41][43][44]
De e mining he Op imal Fo mula o C ude Papain Sc ub C eam.
Moch Fu uchul A i in e al. Fo mula ion o D ied C ude Papain in an Ex olia ing Sc ub C eam: Op imiza ion Using
Mini ab 20 Response Op imize and E alua ion o P o eoly ic Ac i i y ia Release Tes . In . J Med. Pha m. Res., 6 (6):
340‐351, 2025
348
Op imiza ion o he c ude papain sc ub c eam o mula ion was pe o med using Mini ab 20 esponse su ace
me hodology. Fou key pa ame e s— iscosi y, sp eadabili y, pH, and p o eoly ic ac i i y—we e selec ed as c i ical
quali y a ibu es. Each esponse was assigned a a ge ange o guide he op imiza ion p ocess: iscosi y was minimized
o enhance ease o applica ion, sp eadabili y was maximized o ensu e uni o m dis ibu ion, pH was cons ained be ween
3.5 and 8.0 o main ain enzyme s abili y and skin compa ibili y, and p o eoly ic ac i i y was maximized o p ese e
enzyma ic e icacy. Following he es ablishmen o hese a ge s, he esponse op imize ool in Mini ab 20 was execu ed.
The esul ing op imal o mula ion consis ed o 10% s ea ic acid, 4% ie hanolamine (TEA), and 2% ce yl alcohol,
yielding he ollowing p edic ed esponses: Viscosi y o 82,666 cP; Sp eadabili y o 114.97 g·cm/s; P o eoly ic ac i i y
o 4.2403 TU/mg; pH o 7.8533, and Desi abili y index o 0.8518. [45]
These esul s indica e a well-balanced o mula ion wi h high desi abili y, e lec ing he simul aneous achie emen o
mul iple pe o mance c i e ia. The selec ed concen a ions o s ea ic acid and TEA con ibu e o emulsion s abili y and
app op ia e alkalini y, while ce yl alcohol enhances consis ency wi hou excessi ely inc easing iscosi y. The pH alue
alls wi hin he op imal ange o papain ac i i y, which ypically lies be ween pH 6.0 and 8.0, ensu ing enzyma ic
unc ion is e ained. [46]
The desi abili y unc ion app oach used he e is widely ecognized o mul i- esponse op imiza ion in cosme ic and
pha maceu ical o mula ions, allowing o he in eg a ion o di e se pe o mance me ics in o a single p edic i e model.
This me hod suppo s a ional o mula ion design by balancing ade-o s be ween compe ing a ibu es such as ex u e,
bioac i i y, and skin eel. [47]
Papain Release Kine ic Analysis
The elease kine ics o papain om he sc ub c eam o mula ion we e e alua ed by analyzing he cumula i e pe cen age
o papain eleased (M ) o e ime. Fou kine ic models we e applied o de e mine he elease mechanism: ze o-o de ,
i s -o de , Higuchi, and Ko smeye –Peppas. The co esponding equa ions, eg ession coe icien s (R2), and elease
exponen (n) alues a e p esen ed in Table 8.
Based on he eg ession analysis, he Higuchi and Ko smeye –Peppas models p o ided he bes i o he expe imen al
da a, wi h R2 alues o 0.9754 and 0.9745, espec i ely. The elease exponen (n = 0.4768) om he Ko smeye –Peppas
model indica es a Fickian di usion mechanism, sugges ing ha papain elease is p ima ily go e ned by di usion h ough
he c eam ma ix [48].
The Higuchi model u he suppo s his conclusion, as i desc ibes d ug elease om a homogenous ma ix sys em whe e
he a e is p opo ional o he squa e oo o ime. This beha io is consis en wi h he physicochemical p ope ies o he
sc ub c eam, whe e iscosi y and ma ix s uc u e in luence he di usion o papain [49].
These indings a e c i ical o unde s anding he elease dynamics o enzyma ic ac i es in semisolid o mula ions and
op imizing deli e y sys ems o opical applica ions.
CONCLUSION
The s udy success ully o mula ed and op imized a c ude papain sc ub c eam using esponse su ace me hodology and
mul i- esponse desi abili y analysis. Among he es ed o mula ions, Fo mula 2, composed o 10% s ea ic acid, 4%
ie hanolamine (TEA), and 2% ce yl alcohol, demons a ed he mos a o able balance o physicochemical and
unc ional p ope ies:
• Viscosi y: 82,666 cP — sui able o de mal applica ion wi hou comp omising sp eadabili y
• Sp eadabili y: 114.97 mm — ensu ing ease o applica ion and uni o m skin co e age
• pH: 7.85 — wi hin he op imal ange o papain ac i i y and skin compa ibili y
• P o eoly ic ac i i y: 4.24 TU/mg — indica ing e ec i e enzyma ic unc ion
• Desi abili y index: 0.8518 — e lec ing s ong mul i-pa ame e op imiza ion
The indings con i m ha ce yl alcohol concen a ion in e sely a ec s p o eoly ic ac i i y due o inc eased iscosi y,
which limi s enzyme di usion, consis en wi h he S okes–Eins ein di usion heo y. Con e sely, TEA-s ea ic emulsi ie
enhances papain s abili y, likely by imp o ing emulsion in eg i y and p o iding a p o ec i e mic oen i onmen . This
op imized o mula ion o e s a p omising base o cosmeceu ical applica ions, pa icula ly in ex olia ing and
enzyma ically ac i e skinca e p oduc s. The in eg a ion o s a is ical modeling wi h expe imen al alida ion suppo s
a ional design and scalabili y o enzyme-based opical sys ems. The mos ele an mechanism o he elease o papain
om sc ap c eam p epa a ions is a combina ion o Fickian di usion and elaxa ion o he c eam ma ix.
