International Journal of Biological Macromolecules 147 (2020) 973–979

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International Journal of Biological Macromolecules

journal homepage: www.elsevier.com/locate/ijbiomac

Amphiphilic cationic polymers as effective substances improving the

safety of use of body wash gels
Tomasz Bujak, Zofia Nizioł-Łukaszewska, Aleksandra Ziemlewska ⇑
Department of Technology of Cosmetic and Pharmaceutical Products, University of Information Technology and Management in Rzeszow, Kielnarowa 386a, 36-020 Tyczyn, Poland

a r t i c l e i n f o a b s t r a c t

Article history: Cationic surfactants have a wide range of applications in the cosmetic industry. The present study is an
Received 6 August 2019 attempt to determine the effect of an amphiphilic cationic polymer – a cationic derivative of guar gum –
Received in revised form 25 September on the safety of use of body wash cosmetics. Model body wash gels based on an anionic surfactant, con-
2019
taining 0.05, 0.25 and 0.5% of the studied compound, were subjected to analyses in order to evaluate their
Accepted 6 October 2019
Available online 31 October 2019
skin irritation effect and the ability to solubilize model sebum. Cell culture studies were also carried out.
In addition, the effect of the cationized guar gum derivative on the key quality parameters of body wash
cosmetics (viscosity and foaming properties) was assessed. The analyses showed that the addition of the
cationic guar gum derivative induced a significant increase in the safety of use of the model body wash
gels without causing any impairment of the functional properties of the product.
Ó 2019 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://
creativecommons.org/licenses/by/4.0/).

1. Introduction hair surface becomes smoother, and the effect of natural gloss is
seen [1–3,9].
Cationic surfactants have found applications in a number of In addition to binding to the hair surface, cationic surfactants
industries. Due to their specific chemical structure and its resulting have the ability to bind to keratin in the skin. This property has
properties cationic surfactants are used, among others, in the phar- been used by the manufacturers of washing cosmetics by adding
maceutical industry, cosmetic industry, household chemicals, cationic surfactants also to products designed for body and hand
material engineering [1–5], and dyeing processes [6–8]. In aqueous washing. Act of such products rely on the ability of cationic surfac-
solutions, cationic surfactants dissociate to form a surface-active tants to significantly improve their sensory properties. After rins-
cation which can interact with negatively charged surfaces by ing off, the skin appears seemingly moisturized and smooth,
modifying them (hydrophobization). In addition, cationic surfac- which is a result of an interaction between the substances and skin
tants are used as preservatives and disinfectants agents due to surface and proteins, which is analogous to the interaction taking
their bacteriostatic activity [1–5]. place with the surface of hair [9].
The greatest use of cationic surfactants has been observed for Changing trends in the cosmetic market combined with grow-
many years in the cosmetic and household chemical industries. ing awareness of consumers have forced manufacturers to make
These substances are the main active ingredients in a range of efforts towards formulating products with high levels of safety-
products including hair conditioners, hair styling products or con- in-use through the selection of skin-safe washing compounds
ditioning shampoos [1,2]. The products make use of the property of (e.g. alkylpolyglucosides) or the addition of appropriate active sub-
cationic surfactant binding to the negatively charged hair surface. stances (e.g. oils, plant extracts). The use of surfactants of natural
Since electrostatic interactions are strong, cationic surfactants origin and the incorporation of selected active substances usually
remain bound on the surface of hair even after a hair care product result in a decrease in qualitative and functional parameters of
is rinsed off. Electrostatic interactions of cationic polymers with the product, such as washing performance or foaming properties
the hair surface result in the neutralization of charge, which gives [9–11].
this group of compounds an antielectrostatic effect. Also, owing to Therefore, an essential aspect in the formulation of cosmetic
the orientation of the hydrophobic parts and the possible penetra- products is the selection of product ingredients, so that safety-in-
tion of cationic surfactants into defects in the hair structure, the use is enhanced without compromising the quality of the formu-
lated product. Currently, the most popular washing compound
used in body wash cosmetics is the anionic surfactant Sodium
⇑ Corresponding author. Laureth Sulfate (SLES). The popularity of SLES is mainly due to eco-
E-mail addresses: [email protected] (T. Bujak), [email protected] (Z. nomic reasons (relatively low price compared to other surfactants),
Nizioł-Łukaszewska), [email protected] (A. Ziemlewska).

https://doi.org/10.1016/j.ijbiomac.2019.10.064
0141-8130/Ó 2019 The Authors. Published by Elsevier B.V.
This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
974 T. Bujak et al. / International Journal of Biological Macromolecules 147 (2020) 973–979

technological aspects (good performance at low concentrations raw material alternative to substances such as Polyquaternium-7.
and possibility to achieve the desired product consistency – viscos- At present, the substance is used as a conditioning ingredient in
ity – by adding sodium chloride, which also brings economic ben- hair conditioners and 2-in-1 shampoos.
efits), and qualitative aspects (possibility of formulation a product The studies involved the formulation of a series of model wash-
with desired washing performance and foaming properties). A dis- ing systems containing an amphiphilic cationic polymer of natural
advantage in use of this surfactant is related to its relatively high origin – Guar Hydroxypropyltrimmonium Chloride. The model cos-
potential to induce skin irritation reactions by creating strong ionic metics thus obtained were evaluated in order to determine their
bonds with the epidermal proteins [1,2,9,12–17]. safety-in-use (zein test, ability to solubilize sebum, and cell culture
For this reason, cosmetic manufacturers are constantly looking cytotoxicity studies), and identify their qualitative parameters (vis-
for new solutions to reduce the negative impact of SLES on the skin. cosity, foaming properties).
A number of methods to achieve this effect have been elaborated.
For example, the formulations may be enriched with additional
2. Materials and methods
substances such as polymers, proteins and it’s hydrolysates [18–
28], plant extracts and oils [29–33]. Alternatively, delicate surfac-
2.1. Materials
tants or their mixtures may be used (e.g. anionic with nonionic
and amphoteric or cationic types) [1,5,34–38]. Structures arising
Raw materials used in the commercial cosmetics were used to
in such systems, e.g. surfactant and polymer combinations, or the
develop the body wash gels: Sodium Laureth-2 Sulfate (SLES, trade
formation of mixed micelles in systems containing different types
name Brensurf 25; supplier Brenntag, Poland), Cocamidopropyl
of surfactants, decrease the adverse effects caused by washing
Betaine (Dehyton K, BASF, Germany), Citric Acid (Citric Acid, Chem-
compounds on the skin. A frequently used method of reducing
pur, Poland), Guar Hydroxypropyltrimmonium Chloride (Dehy-
the adverse skin effects caused by anionic surfactants is the incor-
quart Guar TC, BASF, Germany), Sodium Chloride (Sodium
poration of a cationic surface-active compound into the product.
Chloride, Chempur, Poland), Sodium Benzoate (Purox S, Brenntag,
Among an extensive range of cationic surfactants, the most
Poland), MiliQ water.
widely used group in the cosmetic industry comprises ammonium
In the physico-chemical tests following were used: zein from
chlorides (e.g. cetrimonium chloride) as well as various types of
corn (Zein, Sigma Aldrich, USA), Albumin (Albumin Bovine fraction
ammonium derivatives referred to as polyquaterniums, or cationic
V, Bioshop, Canada), potassium sulfate (Chempur, Poland), copper
silicone derivatives. Disadvantages of these substances include
sulfate pentahydrate (Chempur, Poland), sulfuric acid 98% (Chem-
their relatively high potential to produce skin and eye irritation,
pur, Poland), Tashiro indicator (Chempur, Poland), sodium hydrox-
and adverse effects on the natural environment. These compounds
ide, citric acid (Chempur, Poland). All reagents were analytical
are classified as toxic to aquatic organisms with long-term effects.
grade.
In addition, they may have a corrosive effect on the skin and cause
eye damage. As a result, a need arose to develop a cationic surfac-
tant without any adverse effects on human health nor to the natu- 2.2. Methods
ral environment [1–5,9].
In the course of development efforts, a special interest was 2.2.1. Development of formulations and technologies for the
focused on the cationic amphiphilic polymer called Guar Hydrox- production of model washing gels
ypropyltrimonium Chloride. It is being obtained from guar gum The formulations of model washing gels are listed in the
(polysaccharide – galactomannan), a substance of natural origin Table 1.
produced from the seeds of the plant Cyamopsis Tetragonoloba Four samples have been prepared for the study. One of them
[39]. The production process of the cationic derivative includes a was the reference sample (base). The other samples contained
quaternization reaction of guar flour with (3-chloro-2- cationic guar gum (KP) at a concentration of 0.05 (KP_0.05), 0.25
hydroxypropyl) trimethylammonium chloride. The chemical struc- (KP_0.25) and 0.5 wt% (KP_0.5). Each samples contained 10 wt%
ture of cationized guar gum is shown in Fig. 1. of surfactants: 8.5% of Sodium Laureth Sulfate (SLES) and 1.5 wt%
Unlike the commonly used cationic surfactants, Guar Hydrox- of Cocamidopropyl Betaine (CAPB). Sodium Chloride (0.5 wt%)
ypropyltrimonium Chloride has no adverse impact on the natural was used as a viscosity modifier and Citric Acid (0.4–0.5 wt%)
environment or human health, and it may serve as a sustainable was used as a pH modifier. The samples were preserved by Sodium
Benzoate.
Production technology of the model washing gels was as follow:
water was weighed into the beaker and then the KP was added. The
solution was mixed with a mechanical stirrer (Chemlad O20,
250 rpm, 5 min). To dissolve the KP, the solution was acidified with
citric acid and stirring was continued (350 rpm, 10 min). Then the
surfactants were added. After surfactants dissolution sodium chlo-
ride and sodium benzoate were added. The samples were mixed
until the ingredients were completely dissolved. In the final step,
the pH of the formulations was adjusted to 5.5 ± 0.1 using 10%
aqueous citric acid solution.

2.2.2. Determination of irritant potential – Zein volume

Irritant potential of the products was measured using zein test.
In the surfactants solution a zein protein is denatured and then is
solubilized in the solution. This process simulates the behavior of
surfactants in relation to the skin proteins.
To 40 mL of the samples a solution (10% wt) was added
2 ± 0,05 g of zein from corn. The solutions with zein were shaken
Fig. 1. Chemical structure of cationic guar gum. in a shaker with water bath (60 min. at 35 °C). The solutions were
 T. Bujak et al. / International Journal of Biological Macromolecules 147 (2020) 973–979 975

Table 1
The formulations of model washing gels.

Ingredient Concentration [wt%]

Base KP_0.05 KP_0.25 KP_0.5
Aqua To 100
Guar hydroxypropyltrimmonium chloride 0 0.05 0.25 0.5
Citric acid 0.05
Cocamidopropyl betaine 1.50
Sodium laureth sulfate 8.50
Sodium chloride 0.50
Citric acid (10% aq.) To pH 5.5 ± 0.1
Sodium benzoate 0.6

filtered on Whatman No. 1 filters and then centrifuged at 5000 rpm 2.2.6. Cytotoxicity
for 10 min. The nitrogen content in the solutions was determined HaCaT cells (human keratinocytes, ATCCÒ) were obtained from
by Kjeldahl method. 1 mL of the filtrate was mineralized in sulfuric the American Type Culture Collection (Manassas, VA 20108, USA).
acid (98%) containing copper sulphate penthahydrate and potas- The keratinocyte culture was established in the DMEM medium
sium sulphate. After mineralization, the solution was transferred (Dulbecco’s Modified Eagle’s Medium, Gibco) containing L-
(with 50 mL of MiliQ water) into the flask of the Wagner–Parnas glutamine, 5% FBS (Foetal Bovine Serum, Gibco) and antibiotics
apparatus. 20 mL of sodium hydroxide (25 wt%) was added. The (100 lg/mL of penicillin and 1000 lg/mL of streptomycin, Gibco).
released ammonia was distilled with steam. Ammonia was bound Cell cultures were maintained at a temperature of 37 °C, at a car-
by sulfuric acid (5 mL of 0.1 N H2SO4) in the receiver of the Wagner bon dioxide concentration of 5%. To perform cytotoxicity tests,
– Parnas apparatus. The unbound sulfuric acid was titrated with cells were seeded into 96-well plates at a density of 10,000 cells/
0.1 N sodium hydroxide. Tashiro solution was used as an indicator. well. Following cell attachment to the bottom of the plates, the
The zein number (ZN) was calculated from the equation: keratinocyte cells were treated with different concentrations
(0.125; 0.25; 0.5; 1%) of the studied model washing systems con-
ZN ¼ ð10  V 1 Þ  100  0:7 ½mg N=100mL taining the amphiphilic cationic polymer Guar Hydroxypropy-
ltrimmonium Chloride at various concentrations.
where V1 is the volume (cm3) of sodium hydroxide used for titra-
tion of the sample. The final result was the arithmetic mean of five
independent measurements. 2.2.6.1. Neutral Red test. The proliferation and metabolism of ker-
atinocytes were determined using the Neutral Red dye (Sigma
Aldrich) according to the procedure described by Borenfreund
2.2.3. Determination of the foaming properties et al. The test involves the uptake of the Neutral Red dye by lyso-
The method of measurement was in line with Polish Standard somes. The dye is capable of penetrating through the membranes
PN-ISO 696:1994P (Surface active agents - Measurement of foam- and accumulating in the lysosomes of exclusively live intact cells
ing power - Modified Ross-Miles method). The foam volume pro- [40]. To assess the effect of the studied extracts, the cells were
duced by 500 mL of samples solutions (1 wt%) falling from a seeded into 96-well plates at a density of 10,000 cells/well. Follow-
height of 450 mm into a cylinder (1000 mL) containing 50 mL of ing cell attachment to the bottom of the wells, the cells were incu-
the same solution was measured. Measurements were carried bated for 24 h with the studied washing systems at varying
out at 22 °C. The final result was the arithmetic mean of five inde- concentrations (0.125; 0.25; 0.5; 1%). The control group consisted
pendent measurements. Foam stability was calculated from the of cells without any exposure to the extracts (cultured in the med-
equation: ium alone). Following 2-hour incubation, the medium containing
the model cosmetics was removed, and a new medium containing
V 10
Foam stability ¼ x100% the Neutral Red dye (40 lg/ml) was added. Next, the cells were
V1 washed with PBS, and a 150 ll portion of destaining solution
(EtOH/AcCOOH/H2O2, 50%/1%/49%) was added to each well. The
where: V10 – foam volume after 10 min, V1 – foam volume after
plates together with the cells were gently shaken for 10 min until
1 min.
the dye was extracted from the cells, and a homogeneous solution
was formed. Cell viability was determined on the basis of optical
2.2.4. Viscosity measurements density (OD) of the eluted dye by measuring absorbance at a wave-
A Fungilab Expert (Fungilab, Spain) rheometer was used. Mea- length k = 540 nm using a microplate reader Filter Max 5 (Thermo
surements were carried out at 22 °C with a rotary speed of the Scientific). The experiments were conducted in triplicate for each
spindle of 10 and 50 rpm. Viscosity values presented in the figures concentration of the extract. The results were expressed as a per-
below represent average values obtained from five independent centage of cell viability determined for the cells treated with the
measurements. studied extracts compared to the control (100%).

2.2.5. Solubility of model sebum 3. Results and discussion

5 g of stearic Acid and 0.5 g of cholesterol was mixed with
100 mL of 5 wt% aqueous solution of samples. The mixutes were 3.1. Ability to solubilize model sebum – Damage to the hydrolipidic
shaken on a shaker with water bath (24 h at 25 °C). After 24 h barrier
the solutions were filtered on Whatman No. 1 filters. The filters
were then washed repeatedly with demineralised water and The ability of surfactants to solubilize and dissolve the compo-
dryied. The amount of solubilised stearic acid was determined nents of the hydrolipidic layer present on the skin surface repre-
grawimetric. The values presented in the figures represent average sents one of the mechanisms potentially leading to skin irritation
values obtained from five independent measurements. due to the use of body wash cosmetics. Sebum solubilizaton
976 T. Bujak et al. / International Journal of Biological Macromolecules 147 (2020) 973–979

Fig. 2. Solubility of model sebum in the solutions of model washing gels. Fig. 3. Irritant potential of model washing gels.

impairs the barrier functions of the skin, and causes an increase in The obtained results show that the addition of cationic surfac-
TEWL value. Consequently, a crucial aspect in the formulation of tants contributes to a significant reduction in the skin irritation
body wash cosmetics is the adjustment of product composition effect of the model body wash gels. Even a low concentrations of
in such a manner as to reduce the capacity of the product to cause the cationized guar gum derivative (0.05%) applied was just
powerful dissolution of fatty ingredients. The results of the study enough to induce an approximately 10% decrease in zein value
evaluating the ability of the test samples to dissolve model sebum (ZV) comparing to the baseline sample. An increase in the concen-
are presented in the figure 2 (see Fig. 2). tration of the cationic surfactant causes a further decrease in the
As demonstrated by the analysis of the results obtained in the value of the parameter under analysis. The result obtained for
study, incorporating a cationic polymer (CP) into the formulations the sample KP_0.5 was about 35% lower relative to the baseline
of the model body wash gels contributes to an improvement in sample (170 mg N/100 mL). According to the literature data
their safety of use. CP addition lowers significantly the sebum dis- [2,3,11] the irritant potential of body wash cosmetics can be esti-
solution ability, and thus decreases the potential of the model body mated on the basis of the determined zein value. If the value is
wash gels to damage the hydrolipidic barrier of the skin. Even a within the range of 200–400 mgN/100 mL, cosmetics are classified
low concentration of CP added to the formulation is sufficient to as moderately irritant to the skin, above 400 mgN/100 mL –
decrease the ability of the body wash gel to dissolve the model strongly irritant, and below 200 mgN/100 mL – non-irritant.
sebum (stearic acid). Compared to the baseline sample, for which Studies conducted to date show that the ZV for 1% SLES solution
the result of 4.2% was obtained, the ability to solubilize fatty sub- is approximately 300–320 mg N/100 mL. Based on the value, this
stances determined in the sample containing 0.05% of CP was anionic surfactant is classified as moderately irritant to the skin
approximately 10% lower. As the concentration of CP in the sample [3,4,43,44]. In practice, various solutions are employed with a view
increased, the results were found to decrease further. The amount to reducing the irritant effect of SLES. The most popular method is
of solubilized sebum determined in the sample with the highest CP the addition of amphoteric or nonionic surfactants. In such sys-
concentration was approximately 25% lower than in the baseline tems, mixed micelles are formed, which are far greater in size
sample. and more stable than micelles built from a single type of surfac-
Research conducted by Ananthapadmanabhan et al. [41,42] tant. Larger-sized micelles have no capacity to penetrate into dee-
shows that the composition of surfactants in the formulation sig- per layers of the epidermis, and hence their interaction with skin
nificantly affects the ability of the system to damage the hydroli- proteins is limited. In addition, they are characterized by greater
pidic layer on the skin surface. Gels containing a high proportion stability, which lowers the release of individual surfactant mole-
of CAPB are characterized by a greater ability to dissolve lipids than cules (monomers) into the bulk phase of the solution. Monomers
the solution of SLES. The results obtained in the reported study are claimed to have the strongest potential to trigger skin irrita-
indicate that the addition of a cationic guar gum derivative con- tions due to the small size of their molecules and high penetration
tributes to a decrease in the solubilization abilities of the system. capacity [5,18,19,22,36,43,45–48]. Another method of reducing the
Consequently, the substance may be an excellent ingredient for adverse impact of surfactants on the skin is the addition of poly-
the formulation of sustainable and safe body wash cosmetics. mers. By forming specific structures with surfactant micelles, poly-
Owing to their ionic (cationic) structure and surface activity, mers stabilize them, lowering the release of monomers [25–28]. As
amphiphilic cationic polymers exhibit an ability to adsorb on the the literature data show [26–28], the measures presented above
surface of sebum. This activity may result in a protective effect of contribute to a decrease in zein value. In systems containing mixed
CP on epidermal lipids [1,37,38]. Consequently, CP addition to for- micelles, zein values are reduced by approximately 10–15% com-
mulations leads to a decrease in the ability of other surfactants to pared to solutions formulated exclusively with an anionic surfac-
dissolve and elute sebum components from the skin. tant, while the addition of polymers (0.5% of
polyvinylpyrrolidone) lowers the ZV of SLES by about 20–25%. As
shown by the analyses, the addition of amphiphilic cationic poly-
3.2. Skin irritation potential mers causes a decrease in ZV to approximately 35%, which demon-
strates that the surfactant is highly effective at lowering the
The potential to trigger skin irritations is one of the greatest dis- irritant effect. In solutions containing a mixture of anionic and
advantages connected with the use of body wash cosmetics. Their amphoteric surfactants, the addition of a cationic surfactant results
constituent surfactants may bind to the epidermal surface pro- in the formation of large-sized mixed micelles, and guarantees
teins, causing their denaturation and then elution from the skin. higher stability of aggregates present in the solution. Moreover,
The highest ability to cause skin irritation is associated with a pres- the polymeric structure of cationic guar gum derivatives addition-
ence of anionic surfactants which bind to proteins via strong ionic ally stabilizes the micelles arising in solutions, which significantly
interactions. The results of measurements of the skin irritation lowers the release of surfactant monomers, and thus strongly
effect are shown in Fig. 3. reduces the skin irritation effect [1,25].
 T. Bujak et al. / International Journal of Biological Macromolecules 147 (2020) 973–979 977

3.3. Cytotoxicity

3.3.1. Neutral Red test

In our cytotoxicity analyzes of cationic surfactants, the HaCaT
line (keratinocytes) was selected as the cell line for the study.
These cells were chosen because they are the first skin cells that
contact surfactants when using cleaning agents. Previous studies
have suggested that cultured human keratinocytes allow us to pre-
dict dermal irritancy caused by several surfactants in human sub-
jects [47–49].
Cytotoxicity was assessed with the Neutral Red test. The dye is
widely used by many researchers as a marker for the evaluation of
cell proliferation and metabolism in various types of compounds
under in vitro conditions. The test was selected because of its sen-
sitivity, speed, safety and cost efficiency. The Neutral Red test is Fig. 4. The effect of the model body wash gels at different concentrations (0.125,
based on the assessment of the ability of viable, intact cells to 0.25, 0.5 and 1%), without the addition of Guar Hydroxypropyltrimmonium
incorporate and bind a neutral red dye in lysosomes [49,50]. Chloride on Neutral Red Dye uptake in cultured keratinocytes cells after 2 h of
exposure. Data are the mean ± SD of three independent experiments, each of which
Wilhelm et al. conducted studies to assess the cytotoxic activity
consists of three replicates per treatment group.
of anionic surfactant SLS on human keratinocytes (HaCaT). In their
studies, they proved that the longer exposure time of cells to the
action of this surfactant reduces their lifespan. They also proved
that as the concentration of SLS increases, the retention index of
neutral red in the lysosomes decreases, which indicates a decrease
in the viability and metabolic activity of the examined cells [49].
Sanchez et al. in their experiments investigated the cytotoxicity
of synthesized surfactants being derivatives of lysine and commer-
cial anionic surfactants by analyzing the integrity of cell mem-
branes (Neutral Red) and metabolic activity (MTT) of human
keratinocytes in vitro. The results of their studies clearly indicate
the toxic effect of all tested surfactants on human keratinocytes.
Similarly to the aforementioned authors, they also showed that
the cytotoxic effect increases with increasing concentrations of
surfactants. The studies also showed that the lysine derivative sur-
factants show a weaker cytotoxic effect compared to the commer-
cial surfactants tested. The authors also observed differences in the
Fig. 5. The effect of the model body wash gels at different concentrations (0.125,
activity of the anionic surfactants studied and indicated that the 0.25, 0.5 and 1%), containing Guar Hydroxypropyltrimmonium Chloride at concen-
weaker cytotoxic effect is exhibited by surfactants with heavy tration 0.05% on Neutral Red Dye uptake in cultured keratinocytes cells after 2 h of
counterions. Nevertheless, they did not found differences between exposure. Data are the mean ± SD of three independent experiments, each of which
surfactants with counterions of the same size. However, the stud- consists of three replicates per treatment group.
ies indicated that the tested anionic surfactants being derivatives
of lysine are even 6 times less toxic than commonly used in SDS
washing preparations. In contrast, HTAB cationic surfactant
showed the strongest cytotoxic effect on keratinocytes in both
tests [51].
The studies evaluated the effect of the model body wash gels at
different concentrations (0.125, 0.25, 0.5 and 1%), containing Guar
Hydroxypropyltrimmonium Chloride at varying concentrations
(0.05, 0.25 and 0.5%), on the proliferation and metabolism of HaCat
cells. Where no cationic polymer was included in the formulation
of the model cosmetic, a drop in the number of viable keratinocyte
cells (even up to 42% in relation to the baseline value in the gel at a
concentration of 1%) was observed (Fig. 4). As shown in the graphs
(Figs. 5–7), the model body wash gels with added of CP induce the
proliferation and metabolism only at the lowest concentrations
(0.125 and 0.25%). A rise in the concentration of the model washing
systems was found to be accompanied by a decrease in the viabil- Fig. 6. The effect of the model body wash gels at different concentrations (0.125,
ity of keratinocyte cells. The highest proliferative capacity (even up 0.25, 0.5 and 1%), containing Guar Hydroxypropyltrimmonium Chloride at concen-
tration 0.25% on Neutral Red Dye uptake in cultured keratinocytes cells after 2 h of
to 154% higher than in the control sample) was demonstrated in
exposure. Data are the mean ± SD of three independent experiments, each of which
the gel with added cationic polymer at a concentration of 0.25% consists of three replicates per treatment group.
(Fig. 4). Based on these results, it can be concluded that the addi-
tion of the cationic polymer Guar Hydroxypropyltrimmonium
Chloride positively affects the proliferation and metabolism of skin 3.4. Analysis of functional properties of model body wash gels
cells under in vitro conditions only at low concentrations (up to
0.25%.) It is assumed that at higher concentrations the model body As already mentioned, a crucial aspect in the formulation of
wash gel will act cytotoxically on keratinocyte cells. body wash cosmetics is to make sure that the measures employed
978 T. Bujak et al. / International Journal of Biological Macromolecules 147 (2020) 973–979

perspective of product application. Higher viscosity at low speeds

facilitates dispensation and prevents the product from flowing
over the hand during use, while a drop in viscosity at higher forces
acting on the sample facilitates product spreadability on the skin.
This behaviour improves product performance, which translates
into a decrease in the quantity of cosmetics entering the sewage
system during a single use. These results support the claim that
cationized guar gum is a cosmetic raw material with potential ben-
efits in the formulation of body wash products with improved
safety-in-use.
The increase in viscosity observed in the samples with added CP
is a consequence of the type of structures arising in the system. In
mixtures containing anionic and cationic surfactants, mixed
micelles are formed. Because of their large size and cylindrical
Fig. 7. The effect of the model body wash gels at different concentrations (0.125, shape, they induce a viscosity increase. The formulations of the test
0.25, 0.5 and 1%), containing Guar Hydroxypropyltrimmonium Chloride at concen- samples contain the same concentration of sodium chloride which
tration 0.5% on Neutral Red Dye uptake in cultured keratinocytes cells after 2 h of causes a rise in viscosity in systems containing anionic surfactants.
exposure. Data are the mean ± SD of three independent experiments, each of which
The observed correlation between increasing viscosity and the
consists of three replicates per treatment group.
addition of CP creates a possibility to adjust the viscosity level as
desired by using an appropriate polymer concentration. This gives
an opportunity to reduce or eliminate the use of sodium chloride,
which will additionally enhance safety-in-use, as sodium chloride
is known to have a drying effect on the skin [1–5].
The incorporation of CP into the formulations induced a slight
reduction in the foaming ability of the samples under study. How-
ever, as the concentration of CP rose, the stability of the resulting
foam was observed to increase. In the sample containing 0.05% of
CP, the stability of foam was about 7%, and in the sample contain-
ing 0.5% of CP about 9% higher than in the baseline sample.
The increase in foam stability and the decrease in foaming abil-
ity in the CP-containing systems are due to a rise in the viscosity of
1% solutions of the samples used in the study. Higher viscosity has
a limiting effect on foam production under the test conditions, but
Fig. 8. Dynamic viscosity of model body wash gels. after the formation of foam, it reduces the removal of the solution
from the aqueous film separating air bubbles within the foam
structure, which contributes to increasing foam stability. Also,
in order to improve the safety-in-use of the product do not the polymer addition stabilizes the gas bubbles, reducing their coa-
adversely affect their quality characteristics. lescence and the process of foam suppression [1–5].
In the course of the studies, two basic parameters characteriz-
ing the quality and functional properties of body wash cosmetics
were evaluated: viscosity and foaming properties. The results are 4. Conclusions
shown in Figs. 8 and 9.
Added to the formulations of the model body wash gels, CP sig- The reported studies show that the cationic guar gum derivative
nificantly improves their rheological characteristics. As the con- under analysis may find applications not only as an effective con-
centration of CP in the formulation rises, an increase in gel ditioning additive, but also as an ingredient significantly improving
viscosity is noted. The lowest values were obtained for the baseline the safety-in-use of body wash products based on anionic surfac-
sample. Dynamic viscosity at the speeds of 10 and 50 rpm varies tants. The substance under the study, added to the formulations
only slightly, and amounts to approximately 700 mPas. Added at of model body wash products even at low concentrations, was
a concentration of 0.05%, CP increases viscosity to approximately shown to significantly reduce their skin irritation effect. At a con-
1,250 mPAS, and at a concentration of 0.5% – to the value of centration of 0.5%, the zein value dropped by approximately 35%
4,200 mPas. Also, the samples under study reveal a favourable compared to the sample not enriched with a cationic polymer. Fur-
change in viscosity correlated with the forces acting on the sample thermore, it was shown that cosmetics formulated with the addi-
(spindle rotation speed). As the parameter rises, the viscosity of the tion of the analyzed substance had a significantly lower ability to
samples declines. This behaviour is not desirable from the elute and solubilize the fatty components of sebum, which may

Fig. 9. Foaming properties of model washing gels: A – foam ability, B – foam stability.
 T. Bujak et al. / International Journal of Biological Macromolecules 147 (2020) 973–979 979

also contribute to reducing the skin irritation effect of body wash [26] T. Bujak, T. Wasilewski, Z. Nizioł-Łukaszewska, Role of macromolecules in the
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