Escolar Documentos
Profissional Documentos
Cultura Documentos
4, 2013
J.Chem.Soc.Pak.,Vol. 35, No. 4, 2013 1096
(Received on 12th November 2012, accepted in revised form 7th March 2013)
Summary: The present study aimed to formulate and subsequently evaluate a topical skin-care
cream (o/w emulsion) from l-ascorbic acid and its derivatives (sodium ascorbyl phosphate and
magnesium ascorbyl phosphate) at 2% versus its vehicle (Control). Formulations were developed by
entrapping it in the oily phase of o/w emulsion and were stored at 8C, 25C and 40C (in incubator)
for a period of four weeks to investigate their stability. In the physical analysis, the evaluation
parameters consisted of color, smell, phase separation, centrifugation, and liquefaction. Chemical
stability of both derivatives was established by HPLC analysis. In the chemical analysis, the
formulation with sodium ascorbyl phosphate was more stable than those with magnesium ascorbyl
phosphate and l-ascorbic acid. The microbiological stability of the formulations was also evaluated.
The findings indicated that the formulations with l-ascorbic acid and its derivatives were efficient
against the proliferation of various spoilage microorganisms, including aerobic plate counts as well
as Pseudomonas aeruginosa, Staphylococcus aureus, and yeast and mold counts. The results
presented in this work showed good stability throughout the experimental period. Newly formulated
emulsion proved to exhibit a number of promising properties and attributes that might open new
opportunities for the construction of more efficient, safe, and cost-effective skin-care, cosmetic, and
pharmaceutical products.
Keywords: Ascorbic acid; Sodium ascorbyl phosphate; Magnesium ascorbyl phosphate; Stability.
Introduction
An emulsion is a complex mixture of two
immiscible phases, with one phase dispersed in
another. The macroscopic separation of the phases is
prevented by the addition of a suitable surfactant [1].
In an emulsion, the therapeutic properties and
spreading ability of the constituents are increased [2].
In fact, the development of liquid-liquid emulsion is a
common practice in the food and pharmaceutical
industries. A system that consists of oil droplets
dispersed in an aqueous phase is called oil-in-water
or O/W emulsion; a system that consists of water
droplets dispersed in an oil phase is called water-inoil or W/O emulsion [3]. While O/W emulsions are
commonly used as water-washable drug bases and for
general cosmetic purposes, W/O emulsions are
widely used as emollients and for dry skin treatment
[4]. Additional value can be conferred to these
formulations by including active ingredients with
specific cosmetic effects. Particularly advantageous
cosmetic emulsion preparations are obtained when
antioxidants are used as active ingredients [5]. Due to
their beneficial and therapeutic properties, l-Ascorbic
acid (Vitamin C) and its derivatives have often been
incorporated in the form of emulsions in recent
*
Centrifugation Test
Table-1: Physical characteristics of F1 and F2, formulations kept at 8 +2C, 25 +2C and 40 +2C.
Fresh
24h
3 day
7 day
14 day
21 day
28 day
F1
F2
F1
F2
F1
F2
F1
F2
F1
F2
F1
F2
F1
F2
8C
Liquefaction
25C
40C
+
+
+
+
8C
W
W
W
W
W
W
W
W
W
W
W
W
W
W
Color
25C
W
W
W
W
W
W
W
W
W
W
W
W
W
W
40C
W
W
W
W
W
W
W
W
YW
W
YW
YW
YW
YW
8C
Phase
25C
separation
40C
+
+
+
+
8C
Centrifugation
25C
40C
+
+
+
+
=No Change; + =Slight Change; PY= Pale yellow; SYW= Soft yellowish white; YW= Yellowish White; Y= Yellow; W= White.
Table-2: Physical characteristics of F3 and F4, formulations kept at 8 +2C, 25 +2C and 40 +2C.
Fresh
24h
3 day
7 day
14 day
21 day
28 day
F3
F4
F3
F4
F3
F4
F3
F4
F3
F4
F3
F4
F3
F4
8C
Liquefaction
25C
40C
8C
PY
YW
PY
YW
PY
YW
PY
YW
PY
YW
PY
YW
PY
YW
Color
25C
PY
YW
PY
YW
PY
YW
PY
YW
PY
YW
PY
YW
PY
YW
40C
PY
YW
PY
YW
PY
YW
PY
YW
PY
YW
SYW
YW
SYW
YW
8C
Phase separation
25C
40C
8C
Centrifugation
25C
40C
=No Change; + =Slight Change; PY= Pale yellow; SYW= Soft yellowish white; YW= Yellowish White; Y= Yellow; W= White.
Chemical Stability
F1
F2
F3
F4
TEWL (g/m2.h)
7
6
5
4
3
2
1
0
0
2
Time (Weeks)
Table-3: Percentages of nondegraded sodium ascorbyl phosphate (F3) and magnesium ascorbyl phosphate (F4)
(n=3) in o/w emulsions at 2.00%.
Formulation
F3
F4
0
100
100
1
91.116.96
99.914.26
2
885.44
99.018.86
Day
3
73.417.82
98.779.23
7
49.738.78
98.116.66
14
36.741.39
98.045.46
28
28.461.26
97.818.06
Table-4: Microbial load of aerobic plate count, Pseudomonas spp, Staphylococcus aureus and Yeast and molds
count of F1, F2, F3 and F4 during 28 days of storage at 25 + 2C.
0
7
a
F1
F2
F3
F4
2.0+0.30
2.02+0.31 a
2.04+0.25 a
2.01+0.27 a
2.43+0.34
2.15+0.37 a
2.18+0.29 b
2.11+0.19 a
F1
F2
F3
F4
<1
<1
<1
<1
<1
<1
<1
<1
F1
F2
F3
F4
<1
<1
<1
<1
<1
<1
<1
<1
Days of storage at 25 + 2C
14
Aerobic plate count
2.55+0.26 b
2.26+0.18 a
2.28+0.17b
2.24+0.15 a
Pseudomonas spp
<1
<1
<1
<1
Staphylococcus aureus
<1
<1
<1
<1
Yeast and molds
1.56+0.15 a
1.48+0.14 c
1. 41+0.16 a
1.29+0.14 b
21
28
b
2.89+0.19
2.88+0.22 a
2.45+0.18 b
2.36+0.15 a
3.13+0.30 c
2.93+0.29 b
2.59+0.18 c
2.33+0.11 a
<1
<1
<1
<1
<1
<1
<1
<1
<1
<1
<1
<1
<1
<1
<1
<1
1.69+0.39 a,b
F1
1.12+0.28 a
1.32+0.22 a
F2
1.14+0.16 a
1.25+0.22 b
1.58+0.27 a
a
a
1.2+0.15
1.52+0.17 a
F3
1.11+0.11
a
a
F4
1.13+0.16
1.17+0.22
1.32+0.27 b
: Standard deviation of three replicates
CFU: Colony -forming units
ac: Averages for different microbial analyses with different letters in the same column are different (P<0.05).
1.8+0.22 a
1.68+0.27c
1.58+0.23 b
1.39+0.27 a
Experimental
Stability Tests
Formulations
F1
3%
1%
1%
3%
0.5%
6%
2%
2%
0.5%
0.5%
81%
Formulations
F2
F3
3%
3%
1%
1%
1%
1%
3%
3%
0.5% 0.5%
6%
6%
2%
2%
2%
2%
0.5% 0.5%
0.5% 0.5%
2%
2%
79%
79%
F4
3%
1%
1%
3%
0.5%
6%
2%
2%
0.5%
0.5%
2%
79%
Physical Stability
The obtained emulsion was submitted to a
set of organoleptic (color, thickness, look, feel) and
physical (creaming and phase separation) analyses.
Centrifugation Tests
Centrifugal tests were performed for
emulsions immediately after preparation. Those tests
were repeated for emulsions after 24 hours, 7 days,
14 days, 21 days, and 28 days of preparation. They
were performed at 5000 rpm and 25C for 10 minutes
by placing 10g of the sample in centrifugal tubes.
Skin Measurement (Measurement of Transepidermal
Water Loss)
The transepidermal water loss (TEWL) is
determined using skin biophysical technique,
Tewameter CM 210 from Courage1Khazaka
(Cologne, Germany). TEWL, which is related to skin
barrier function, was expressed as g/m2. h [31].
Chemical Stability
All samples were stored in well-closed 25
ml glass flasks. During storage samples were kept at
room temperature (221 C) in the dark, except for
those used for studying the influence of light. The
Staphylococcus aureus
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