Formulation and Evaluation of some Cosmetic preparations using novel natural colorant from Ixora coccinea

Kavyashree Ghongade¹, Vishal Bodake¹, Snehal Badadare², Madhuri Magdum⁴, Nutan Gawande¹, Sanika Kate¹, Karishma Waghmare³

¹Department of Pharmaceutical Chemistry, Gourishankar Institute of Pharmaceutical Education and Research

(B. Pharm.), Limb, Satara.

²Department of Pharmaceutics, G.E.S College of Pharmacy (D. Pharm) Limb, Satara.

³Department of Pharmaceutical Chemistry, Shri D.D. Vispute College of Pharmacy and Research Center,

New Panvel.

⁴Samta Hospital, Dombiwali (E), Mumbai.

*Corresponding Author E-mail: kavyashreeghongade@gmail.com

ABSTRACT:

The present work aimed to formulate various cosmetic formulations comprising of extract of Ixora coccinea (Raktaka). As a natural coloring agent to overcome the drawbacks associated with synthetic colorants, since the synthetic coloring agents may cause allergic reactions and are reported to be carcinogenic. Dried and powdered flowers of Ixora coccinea produces red color. The obtained color was used as a natural colorant in the preparation of different cosmetic preparations namely Lipstick, Rouge, and Eye shadow. Different ingredients such as Bees wax, Carnauba wax, White soft paraffin, castor oil, rice starch, floral essence, and lemon juice etc were used to formulate the aforesaid cosmetics using the extracted color. The prepared formulations were evaluated for various evaluation tests such as color, texture, pH, perfume stability, melting point, breaking point, softening point, surface anomalies, aging stability, thixotropy, skin irritation test depending upon its type. The results of evaluation tests revealed that melting point of the prepared lipstick formulation were observed in the range of 62-64⁰C. The breaking point of all the prepared lipstick formulations noted in the range of 31 – 34gm. Force of application was noted to be good for all the formulations. Thixotropy character was observed in the range of 8.89 to 9.69. Softening point was found to be in the range 52 to 60⁰C. Eye shadow and rouge shows good adhesion property, proper color dispersion, with melting point 49⁰C patch test on volunteer showed no signs of skin irritation. The results of the study verified that the color extracted from the dried and powdered flower of Ixora coccinea may prove to be a beneficial choice and work as an alternative for the synthetic colorants in various cosmetic formulations, which are known to cause allergic reactions and are reported to be carcinogenic.

KEYWORDS: Herbal cosmetic, Ixora coccinea, Lipstick, Rouge, Eye shadow.

INTRODUCTION:

Now-a-days the usefulness of herbs in the Cosmeceuticals production has been extensively increased in personal care system and there is a great demand for the herbal¹. Cosmetic products are used to protect skin against exogenous and endogenous harmful agents and improve the beauty and attractiveness of skin^2,3. The herbal ingredients present in skin care products that supports the strength to the skin, integrity of skin and texture, moisturizing, maintaining elasticity of skin by reduction of collagen and photo protection etc. The usage of synthetic products becomes very harmful from long time for the youth as well as our environment.^4,5

Lipstick is cosmetic preparation used to color the lips and protect them from external environment. Presently the use of this product has increased and choice of shades of colors, textures, luster, have been changed and become wider.^6,7 Rouges are designed to enhance rosy cheek color. The color can vary from the palest of pinks to the deep blue reds but the conventional rouge contains a high proportion of red or reddish brown pigments. Eye shadow is cosmetic which is applied primarily on the upper eyelids and under the eyebrows or around eyes to enhance their brilliance, to help make wear’s eyes stand out look more attractive.

The dyes that contribute to the color of the cosmetics are very harmful to humans on consumption.^8,9 Coal tars are the basic ingredients from which synthetic dyes are formed can cause allergy, nausea, dermatitis, and drying of the skin. In a more ever form they can be carcinogenic and even fatal^10,11,12. Thus, the use of herbal colorants in the cosmetic preparations will surely help to overcome the side effects associated with the synthetic colorants. The present study was therefore undertaken to prepare and evaluate various cosmetics using natural colorant from Ixora coccinea.

Ixora coccinea. is a flowering ornamental plant belonging to family Rubiaceae, is also known as Raktaka, Rukmini/Tetti. It is an important medicinal plant with several medicinal uses in traditional medication system. Raktaka are used in curing various ailments such as dysentery, leucorrhoea activity and anti-inflammatory. Ixora coccinea have therapeutic potential due to presence of bioactive compounds like flavones, isoflavones, flavonoids, anthocyanins, coumarins, lignans, catechins, isocatechins, alkaloids, tannin, saponins and triterpenoids^11,18,19.

MATERIALS AND METHODS:

Materials:

Bees wax, Castor oil, Carnauba wax, White soft paraffin, Cetyl alcohol, Gum acacia were purchased from Loba chem. Floral essence was purchased from local market of Mumbai. Flowers of Ixora coccinea were collected from region of Dapoli. The plant material was authenticated from Department of Botany, YC College of Science, Satara (MS).

Isolation/Extraction of color pigments:

Dried and powdered flowers of Ixora coccinea produces red color. The leaves of Ixora coccinea. and Ixora coccinea were collected from the areas nearby Dapoli region. The flowers were washed carefully using distilled water and shade dried. The shade dried coarsely powdered flowers of Ixora coccinea. (50gm) was taken and the color pigment was extracted by maceration with ethanol (75ml). After overnight maceration, the extract was filtered through filter paper. The extract was concentrated by vacuum distillation to reduce the volume to 1/10, the concentrated extract was transferred to 50ml beaker and the remaining solvent was evaporated on a water bath. Dark reddish colored liquid extract (2.34% w/v) was obtained which was carefully stored in an airtight container till further use.

Preparation of lipstick formulation:

The lipstick was formulated as per the general method. In brief; all waxes namely beeswax, Carnauba wax and soft paraffin were melted together in a porcelain dish on a water bath with a decreasing order of their melting point. The extracted color pigment was mixed with Castor oil and heated. Both the phases were mixed together at the same temperature. Floral essence and lemon juice were added at 40^oC. Then mixture was poured into lipstick mould in excess amount and the mould was kept on an ice bath. After solidification the surplus amount was scrapped off using a knife. Lipsticks were removed from the mould and flamed. Prepared lipsticks are shown in Figure 1

The prepared lipsticks were fitted in a lipstick container and used for further evaluation⁶. As shown in (Table 1).

Table 1. Formulation table herbal lipstick

Ingredients	Role	Quantity taken (gm)
Ingredients	Role	F1	F2	F3	F4	F5
Bees wax	Stiffening agent	8.0	8.0	8.0	8.0	8.0
Carnauba wax	Stiffening agent	4.0	4.0	4.0	4.0	4.0
White soft paraffin	Soft wax (Glossy)	10.0	10.0	10.0	10.0	10.0
Castor oil	Blending agent	2.0	2.0	2.0	2.0	2.0
Color pigment	Coloring agent	-	1.0	1.5	2.0	2.5
Floral (fragrance)	Flavoring agent	q.s	q.s	q.s	q.s	q.s
Lemon juice	Antioxidant	q.s	q.s	q.s	q.s	q.s

Preparation of rouge:

Perfume was mixed properly with starch and the mixture was kept covered for half an hour. The remaining ingredients were mixed thoroughly and sieved through fine muslin. The starch perfume mixture and required color were added to this mixture and stored in a suitable container. Shown in figure 2 and Table 2

Table 2. Formulation table for Rouge

Ingredients	Quantity taken (gm)
Zinc stearate	14.5
Rice starch	14.5
Talc	55.0
Pigment	14.0
Perfume	2.0

Preparation of eye shadow:

Pigment color was added to zinc oxide and white petroleum. The mixture was milled through an ointment mill. Beeswax, spermaceti and lanolin were melted. The color base and perfume were added to it and the final mixture was milled to obtain a uniform product¹⁶. As Shown in figure 3 and Table 3

Table 3. Formulation table for Eye shadow

Ingredients	Quantity (%)
Color pigment	12.0
Zinc oxide	31.0
White petroleum	45.0
White beeswax	5.0
Spermaceti	4.0
Lanolin	4.0
Perfume	q.s.

Evaluation of lipstick formulation:

It is very essential to maintain a uniform standard for lipstick, keeping this view in mind the formulated lipstick was evaluated on the parameters such as melting point, breaking point, force of application, surface anomalies etc^15,16.

Melting point:

Determination of melting point is important as it is an indication of the limit of safe storage. The melting point of formulated lipstick was determined by capillary tube method. The capillary was filled, kept in the capillary apparatus and observed the product for its melting. The procedure was performed for three times and the melting point ratio was observed for the prepared formulations.

Breaking point:

Breaking point was performed to determine the strength of lipstick. The lipstick was held horizontally in a socket ½ inch away from the edge of support. The weight was gradually increased by a specific value (10gm) at specific interval of 30 second and the weight at which it broken was considered as the breaking point.

Force of application:

It is test for comparative measurement of the force to be applied during its application. A piece of coarse brown paper was kept on a shadow graph balance and lipstick was applied at 45º angle to cover a 1 sq. inch area until fully covered. The pressure reading is an indication of force of application.

Surface anomalies:

This was studied by the surface defects, such as no formation of crystals on the surfaces, no contamination by moulds, fungi etc.

Aging stability:

The product was stored at 40ºC for a period of 1 hour. Various parameters such as bleeding, crystallization of the surface and ease of application were observed.

pH parameter:

The pH of the formulated lipstick was determined using pH meter.

Perfume stability:

The lipstick formulation was tested after 30 days, to record the fragrance^14,20.

Softening Point:

Lipstick should be able to withstand range of conditions to which it will be subjected in the consumer’s handbag. It should be resistant to varying temperature conditions and be just as easy to apply in hot as well as in cold weather. Softening point of lipstick was determined by Ring and Ball method. The results are presented in (Table 4).

Ring and Ball method:

A ring or support orifice was taken and prepared herbal lipstick was inserted into it. Extra mass above and below the orifice was removed using a sharp blade leaving a tablet of lipstick fitted into the ring. This was placed in a refrigerator (6°C) for about 10 min. Ring was tied onto a stand. A beaker containing 500mL water at room temperature was placed on a hot plate with magnetic stirrer. A steel ball was delicately placed on the lipstick tablet. The bar with support was then inserted into the beaker till it submerged into it. Heating and slow agitation was then started. Temperature was monitored using a thermometer. The temperature at which the lipstick mass and steel balls were loosed and falls to the bottom of the beaker was noted as softening point of lipstick.

Color and Texture:

Formulated lipsticks were checked for color, glossy and smooth texture ^6.

Solubility test:

The formulated lipstick was dissolved in various solvents to observe the solubility.

Thixotropy character:

It is indication of thixotropic quality and was performed by using penetrometer. A standard needle of specific diameter was allowed to penetrate for 5 seconds under a 50gm load at 25ºC. The depth of penetration was a measurement of the thixotropic structure of lipstick⁴.

Evaluation of rouge:

Color

Formulated rouge was checked for color.

Color dispersion:

This test was performed using a microscope.

Pay-off test:

Pay-off means adhesion characteristic with powder puff were tested on the skin¹¹.

Evaluation of eye shadow:

Color:

Formulated eye shadow was checked for color.

Melting point test:

It is determined by capillary tube for good application and storage point of view.

Texture:

Texture of the eye shadow was checked.

RESULTS:

Evaluation of lipstick:

It is very essential to maintain a uniform standard for herbal lipstick, keeping this view in mind the formulated lipstick was evaluated on the parameters such as melting point, breaking point, force of application, surface anomalies as shown in (Table 4).

Melting point:

The melting point of formulated lipstick was determined by capillary tube method. The melting points of different formulations are shown in (Table 4)

Breaking point:

Breaking point was done to determine the strength of lipstick. The prepared lipstick formulation was held horizontally in a socket ½ inch away from the edge of support. The weight was gradually increased by a specific value (10gm) at specific interval of 30 second and the weight at which it breaks was considered as the breaking point. It was repeated for 3 times and the values are given in (Table 4)

Force of application:

It is the test for comparative measurement of the force to be applied for application of lipstick formulation. A piece of coarse brown paper was kept on a shadow graph balance and lipstick was applied at an angle of 45º to cover a 1 sq. inch area until fully covered. The pressure reading is an indication of force of application as shown in (Table 4).

Surface anomalies:

It was studied by the surface defects, such as no formation of crystals on the surfaces, no contamination by moulds, fungi etc. No defect (s) was observed on the surface.

Aging stability:

The product was stored at 40ºC for a period of 1 hour. Various parameters such as bleeding, crystallization of surface and ease of application were observed.

Solubility test:

The prepared lipstick formulation was dissolved in various solvents to observe its solubility.

pH parameter:

The pH of the formulated lipstick was determined using pH meter. It was performed for 3 times and the values are presented in (Table 4)

Perfume stability:

The formulation of lipstick was tested after 30 days, to record the fragrance. (Table 4)

Softening Point:

Lipstick should be able to withstand range of conditions to which it will be subjected in the consumer’s handbag. It should be resistant to varying temperature conditions and be just as easy to apply in hot and as in cold weather. The softening point of the formulated lipstick was determined by Ring and Ball method. The temperature at which ball looses and falls is noted in (Table 4)

Color and Texture:

Formulated lipsticks were checked for color, glossy and smooth texture.

Thixotrophy character:

Table 4. Evaluation of herbal lipstick

Evaluation parameters	Formulation Batches
Evaluation parameters	F1	F2	F3	F4	F5
Melting point	63 ±2⁰C	62 ±2⁰C	62 ±1⁰C	62 ±1⁰C	64 ±1⁰C
Breaking point	31 ±10gm	34±10 gm	32 ±10 gm	32 ±10 gm	32 ±1.73 gm
Force of application	Good	Good	Good	Good	Good
Surface anomalies	No defect	No defect	No defect	No defect	No defect
Aging stability	Smooth	Smooth	Smooth	Smooth	Smooth
Solubility test	Chloroform	Chloroform	Chloroform	Chloroform	Chloroform
Skin irritation test	No	No	No	No	No
Perfume stability	+++	+++	+++	+++	+++
Softening point	53.66 ±2.08⁰C	56.66 ±1.50⁰C	52.33 ±2.08⁰C	56.66 ±1.50⁰C	59.66 ±1.50⁰C
Color	White	Pale red	Crimson red	Scarlet red	Raspberry red
Texture	Smooth	Smooth	Smooth	Smooth	Smooth
Thixotrophy character	8.89 ±0.15	9.3 ±0.33	9.06 ±0.05	9.16 ±0.30	9.69 ±0.4

Values are mean ± SEM of triplicate determinations

Evaluation of rouge:

Color:

Formulated rouge was checked for color. The observed rouge was cream in color.

Color dispersion:

This test was done by using a microscope. Color particles size was found to be less than 50 um.

Pay-off test:

Pay-off means adhesion characteristic with powder puff should be tested on skin. The results are shown in (Table 5).

Table 5. Evaluation of rouge

Evaluation parameters	Inference
Color	Red
Color dispersion	Properly distributed
Pay-off test	Good adhesion

Evaluation of eye shadow:

Color:

Formulated eye shadow was checked for color.

Melting point:

It was determined by capillary tube method for good application and storage point of view. Melting point of eye shadow was found to be 49⁰C.

Texture:

Texture of the eye shadow was smooth. Shown in (Table 6)

Table 6. Evaluation of eye shadow

Evaluation parameters	Inference
Color	red
Melting point	49.33±1.164 ⁰C
Texture	Smooth

Values are mean ± SD of triplicate determinations

Patch test evaluation of volunteers:

Skin irritation test:

Patch test was performed to check the safety of the formulation and base on human skin. The prepared formulations and base were applied on the forearms of volunteers for 48 h. The values obtained are shown in (Table 7). It was observed that the parameters namely; ease of application, spreadability, sense just after application and on long term, irritation as well as sense on softness on application of formulations A, B, C, D and E over forearms of volunteers, were quite good. Volunteers reported that there was no irritation and redness after application of the prepared formulations. Results of the Patch test are shown in (Figure 4).

Table 7. Evaluation of the prepared formulations on skin (Patch test) in human volunteers

Parameters	Average points for A ± SEM	Average points for B ± SEM	Average points for C ± SEM	Average points for D ± SEM	Average points for E ± SEM
Ease of application	3.4±0.2666	3.2±0.2905	3.4±0.2211	7.5±3.9503	3.4±0.1633
Spread ability	2.7±0.2333	4±0.2582	3.3±0.2134	3.7±0.1527	3.6±0.2211
Sense just after application	3.4±0.400	4.1±0.2768	3.9±0.2768	3.8±0.2494	3.2±0.1333
Sense on Long term	3.6±0.3055	3.9±0.1795	3.8±0.2494	3.8±0.2905	3.6±0.1633
Irritation	4.1±0.1895	4.2±0.2905	4.1±0.2333	4.3±0.2134	4.3±0.2134
Sense of softness	2.8±0.2433	4.2±0.2494	4.3±0.1522	4.3±0.1527	3.9±0.2768

Values are mean ± SEM of determinations (n=10)


Fig 1. Formulated lipsticks (F1, F2, F3, F4, F5)	Fig 2. Formulated rouge	Fig 3. Formulated eye shadow

Fig 4. Patch test of all formulations A) Lipstick base B) Lipstick formulation (F5) C) Rouge formulation D) Eye shadow base E) Eye shadow formulation

(A: Lipstick base, B: Lipstick formulation (F5), C: Rouge formulation, D: Eye shadow base, E: Eye shadow formulation)

DISCUSSION:

With an inherent desire to look beautiful, the recent few years witnessed a marked increase in the use of different cosmetic formulations by women. The demand of various cosmetics is on rise and ever increasing. Varieties of cosmetics from different manufacturers are available in the market. Mostly the preparations contain synthetic ingredients and colorants. Very recently the deleterious effects of such agents have grabbed the attention of the users. And therefore the demand of cosmetics with herbal ingredients is on a rise, as many of these ingredients are devoid of side effects and safe to use. Thus, in the current investigation an attempt was made to formulate and evaluate various cosmetic formulations namely Lipstick, Rouge and Eye shadow using colored pigment from the extract Ixora coccinea. Five formulations of lipstick were formulated and evaluated. The results of the evaluation tests exhibited that the melting point of the prepared lipstick formulation ranged from 62-64^oC, whereas the breaking point of all the prepared lipstick formulations was observed in the range of 31–34gm. The force of application for the prepared formulations was determined and noted to be good. The thixotropic character was noted in the range of 8.89 to 9.69. The Softening point was observed to be in the range of 52.3-59.66⁰C. Different red shades of lipstick formulations namely Crimson red, Scarlet red and Raspberry red were prepared by using varying concentration of colored pigment ranging from 1 to 2.5gm. Amongst the prepared lipstick formulations F5 formulation which comprised of the highest concentration of colored pigment (2.5gm) was subjected to patch test. With the aid of colored pigment from the extract of Ixora coccinea., we have prepared Rouge and Eye shadow preparations. The formulated cosmetics were subjected to different evaluation tests. The results of the evaluation tests revealed that the prepared rouge formulation exhibited proper distribution of color along with good adhesion property as noted in the payoff test. The eye shadow preparation showed pale red color with a melting point of 49⁰C and exhibited smooth texture. Moreover, the results of patch test demonstrated that the prepared cosmetics namely lipstick, eye shadow and rouge formulation showed no signs of skin irritation. Thus, the cosmetic formulations namely lipstick, rouge, and eye shadow were successfully prepared and evaluated by using herbal colored pigment from the extract of Ixora coccinea.

CONCLUSION:

The results of the present investigation revealed that the colored pigment from the flower extract of Ixora coccinea can be effectively used as a natural colorant in various cosmetic formulations. To the best of our knowledge it was the first attempt of wherein we have employed colored pigments from the extract of Ixora coccinea as natural colorants in different cosmetic formulations. The use of a natural colorant will provide a suitable alternative for synthetic colorants which are known for their side effect (s). Thus, this colorant which has been derived from the natural origin can be effectively used in various cosmetic preparations and the side effects associated with the use of synthetic colorants can be efficiently minimized.

ACKNOWLEDGEMENTS:

Authors are thankful to Gourishankar Institute of Pharmaceutical Education and Research limb Satara.

AUTHOR CONTRIBUTIONS:

K.D.G. and V.B. designed the work, performed mathematical and statistical analyses, and wrote the article; N.G. and S.K. carried out cosmetic preparation and evaluation experiments; S.S., M.M. and K.W. critically revised the manuscript.

CONFLICT OF INTEREST:

No conflict of interest was declared by the authors.

REFERENCE:

1. Mali A, Karekar P, Yadav, A Formulation and evaluation of multipurpose herbal cream. Int J Sci Res, 2013, 4(11), 1495-1498.

2. Talpekar P, Borikar M. Formulation, development and comparative study of facial scrub using synethetic and natural exfoliant. Research J. Topical and Cosmetic Sci. 2016, 7(1), 01-08.

3. Gavarkar P, Adnaik R, Chavan D, Bagkar A, Bandgar R. Physicochemical Investigation of Some Marketed Herbal Hair Oil. Research J. Topical and Cosmetic Sci. 2016, 7(2), 70-72.

4. Kumar A, Kumar P, Radha JB, Murli KB, Formulation and evaluation of stable aqueous extract of polyherbal multipurpose face cream. Research J Topical Sci, 2017, 8, 20-18.

5. Kushwaha A, Bhowmick M, Rathi J, Formulation development and evaluation of polyherbal hydrogel for effective treatment of acne. Research J. Topical and Cosmetic Sci, 2017, 8, 1-11

6. Bhagwat D, Patil N, Patel G, Killedar S, More H, Formulation and evaluation of herbal lipstick using lycopene extracted from Solanum lycopersicum L. J Pharm Tech, 2017, 10(4), 1060.

7. Gurubarik N, Saroj R, Jami K, Saravakota S. Formulation and Evaluation of Sun Protection Factor of Poly Herbal Sunscreen Cream. Research J. Topical and Cosmetic Sci. 2016, 7(1), 09-14.

8. Roy A, Ram K. Formulation and Development of Herbal Sunscreen Cream. Research J. Topical and Cosmetic Sci. 2014, 5(1), 12-14.

9. Gurubarik N, Saroj R, Jami K, Saravakota S. Formulation and Evaluation of Sun Protection Factor of Poly Herbal Sunscreen Cream. Research J. Topical and Cosmetic Sci. 2016, 7(1), 09-14.

10. Kothari R, Shukla B, Gautam D, Bagaria M, Sharma A, Formulation and evaluation of herbal lipstick from natural edible coloring matter. Int J The App Sci, 2018, 10(1), 17-20.

11. Harsha S, Verma VS, Sharma M, Singh A, Sharma G, Majumdar M, Gupta S, Sahu SV, Dewangan K, Tripathi DK, Ajazuddin AA. Natural Humectants in Formulation of Calamine Lotion: Its Evaluation and Comparison. Research J. Topical and Cosmetic Sci. 2016, 7(2), 41-45.

12. Roy A, Ram K. Formulation and Development of Herbal Sunscreen Cream. Research J. Topical and Cosmetic Sci. 2014, 5(1), 12-14.

13. Bindu Gopalkrishnan, Roy Chiranjeev. Pharmacognostical Study of Ixora coccinea Flower. Pharmacogn J. 2018;10(5):1042-6.

14. Rajpurohit J, Shaikh S, Valvi P, Jain V, Pawar S, Formulation and evaluation of herbal lipstick by using Bixa orellana as coloring pigment. Int J Pharma Chem Res, 2017, 3, 451-455.

15. Kawarkhe PR, Deshmane SV, Biyani K. Formulation and Evaluation of Antioxidant Face Cream Containing Raspberry Fruit and Grape Seeds Extract. Research J. Topical and Cosmetic Sci. 2016, 7(2), 73-78.

16. Elumalai A, Chinna M, Nikhitha M. Formulation and Evaluation of a Herbal Lipstick from Punica granatum Fruit Peel. Res. J. Topical and Cosmetic Sci. 3(1), 2012, 20-22.

17. Rautela S, Tailor C, Badola A, Formulation and evaluation of a herbal lipstick: A new approach. Int J P’ceutical Erudition, 2013, 3, 26-30.

18. Kalita S, Kumar G, Karthik L, Rao K A Review on medicinal properties of Ixora coccinea. Res J Pharm Tech, (2012), 5(6), 711-714.

19. Nital Patil, Ajit Datar, Extraction, Stability and Separation of Anthocyanins of Ixora coccinea Linn. I J Pharma Pharmac Sci. 2015;7(3):198-202. D

20. Jadhav Kishan, Priyanka Kapare, Divya Khairmode, Nayana Vhatkar, Shraddha Raut, and Akash S. Mali. Development of cosmeceutical cream for dermatitis and acne vulgaris. Res J Topical Cosm Sci. 2018, 9, 33-36.

Received on 21.07.2020 Modified on 20.08.2020

Asian J. Res. Pharm. Sci. 2021; 11(1):22-28.

DOI: 10.5958/2231-5659.2021.00004.7