INTRODUCTION:

Curcuma mangga Val. is a plant that has a lot of benefits for human health, and commonly used for traditional medication. There are so many reports showed that C. mangga has big potency as anti cancer¹, anti-microbe², chemopreventive activity³, as anti-oxidant and anti-diabetic drug⁴ and also influence the estrogen-progesterone profile in rats⁵. However, the usage of the plant extract is inconvenience if it is directly used as medicine especially for topical dosage form. Therefore, a dosage form is needed to be prepared to deliver the plant extract for human usage.

Nanoemulsion is one of the dosage forms which is widely studied particularly for its usage as transdermal dosage form due to the small particle size⁶.

Nano size is mostly considered can increase the bioavailability of active ingredient in the nanoemulsion⁷. However, the nanoemulsion with low viscosity has tendency to be inappropriate for human skin application⁸. Therefore, the addition of gelling agent to the nanoemulsion to produce nanoemulgel is expected to create nanoemulsion with better effect. The nanoemulgel formulation will produce the dosage form that has both nanoemulsion and gel characteristic and will increase the drug permeation into the skin⁹.

Castor oil is one of the vegetable oils that successfully produces nanoemulsion for some drugs such as diclofenac sodium¹⁰ and also increases the permeability of transdermal drug like flurbiprofen¹¹. In this study, MCT oil was combined with the castor oil to produce nanoemulgel with better oil composition and can increase the dosage form stability.

Based on those particular backgrounds, the main objective of this study was to develop a nanoemulgel dosage form containing ethanol extract of C. mangga using Carbopol 940 as gelling agent and to evaluate the physical characteristic and stability of the nanoemulgel for 30 days and also after accelerated test.

MATERIAL AND METHODS:

Materials:

C. mangga was collected from Langkat District, North Sumatera, Indonesia. MCT oil (Okusi Biotech), Castor oil, Tween 80, Ethanol 96% (Merck), Propylene glycol, Tri ethanol amine (TEA), Carbopol 940, and distilled water were purchased from Smart Lab Company, Medan, Indonesia. All chemical reagents used were analytical grade and applied without further purification.

Nanoemulsion Preparation:

The nanoemulsion was prepared by using castor oil and MCT oil (1:1) as oil phase. Tween 80 as surfactant, ethanol and propylene glycol as co-surfactants. The preparation of nanoemulsion was done based on Solans with slight modification¹². Tween 80 was diluted in distilled water and mixed at 60^oC. Castor oil and MCT oil with C. mangga ethanol extract (0.02% for each formula) was mixed in other beaker at 60^oC. The oil phase was added slowly to the water phase, and then the propylene glycol was added and stirred at 1200rpm for 6 hours. The composition of the formula can be seen in the Table 1.

Table 1: Composition of Nanoemulgel of Ethanol Extract C. mangga Formula

Materials		F 1 (%)	F 2 (%)
Oil Phase (4%)	Castor oil	2	2
	MCT Oil	2	2
Surfactant	Tween 80	35	35
Coo-surfactant	Ethanol	15	15
	Propylene Glycol	10	10
	Distilled water	11	11
Gel Base (25%)	Carbopol 940	1.5	0.5
	TEA	3 drops	3 drops
	Distilled water to	25	25

Nanoemulgel preparation:

The preparation of gel base was done by dissolving Carbopol 940 in distilled water with three drops of TEA. The mixture was stirred until a homogenous and transparent gel was obtained. The gel base was left for 24hours. The nanoemulgel of C. mangga ethanol extract was prepared by adding the nanoemulsion to the gel base and homogenized by using high speed homogenizer with 1500rpm speed for 10 min.

Organoleptic test:

Some physically parameters such as consistency, color, odor and visual phase separation were evaluated after preparation and also for 1 month of storage.

Physico-chemical evaluation of nanoemulgel:

pH determination:

pH determination was conducted using a calibrated pH meter instrument. Calibration was done by using a standard buffer solution pH 4 and 7. pH determination was done triplicates for each sample and at room temperature¹³.

Viscosity test:

The viscosity of the nanoemulgel was measured using Brookfield viscometer instrument.

Surface tension determination:

The determination of surface tension was conducted by using Du Nouy tensiometer. The tensiometer was first calibrated using distilled water (theoretical surface tension = 72.75dyne/cm) and the correction factors is the result for theoretical value divided by results obtained.

Type of emulsion test:

The type of emulsion was tested by coloring method using methylene blue. An amount of sample was put on the object glass and mixed with methylene blue and covered with the cover glass. The continuous blue phase due to methylene blue dissolved in water indicates oil in water emulsion type¹⁴.

Homogeneity test:

The homogeneity of nanoemulgel was evaluated by putting an amount of sample on an object glass covered by the cover glass. The nanoemulgel should show no coarse particle detected.

Particle Size Determination:

The globule size of nanoemulgel was measured using particle size analyzer (Horiba instrument LB-550).

Stability Test in Room Temperature:

The nanoemulgel was stored at room temperature and the determination of pH and viscosity value were recorded for 1 month.

Accelerated Stability test:

The stability of the nanoemulgel was evaluated using centrifugation, the heating cooling cycle and shaking tests. The centrifugation test for nanoemulgel was done by putting 10ml of nanoemulgel in the centrifugation tube and spun at 3500rpm for 30 min. The heating cooling cycle test was conducted based on modification method from Khan¹⁵ by putting the nanoemulgel in refrigerator at 4^oC for 12 hours, then continue at room temperature for 12 hours. This step was repeated for 5 cycles. The shaking test was done using orbital shaker with 100rpm for 4 hours. All of the nanoemulgel was evaluated if any phase separation, pH and viscosity value before and after the test.

Data analysis:

All of the data obtained were presented as mean± standard deviation.

RESULTS AND DISCUSSION:

All of the formula in nanoemulsion preparation produced a transparent nanoemulsion and the adding process of gelling agent Carbopol 940 produced a transparent nanoemulgel without any separation, homogenous without any undesirable particle. Higher Carbopol 940 concentration will produce more viscous nanoemulgel. The evaluation for organoleptic characteristic for one month at room temperature can be seen in Table 2.

Carbopol become one of the chosen gelling agents with a lot of advantages in gel preparation. This polymer shows no irritation and toxic effect after repeated use to the skin. For pharmaceutical dosage form, Carbopol is easy to use because it can form gel at room temperature. Carbopol concentration will affect the physical properties of gel and impact the drug absorption and it is suitable for control release purposes¹⁶. Carbopol has an acidic property and it need to be neutralized by alkylating agent such as NaOH or TEA to form gel with a good stability¹⁷. In this study, we used TEA as the neutralizing agent for the Carbopol to form gel. However, acidic characteristic of the extract affected the viscosity of the formed gel.

The physico-chemical evaluation of nanoemulgel was conducted to determine the physical characteristic of the nanoemulgel. The physical characteristic of the nanoemulgel were presented in Table 3. As shown in Table 3, the pH for both formulas were below 6. This pH was still acceptable because the pH balance for skin is in the range of 4.5- 6.5. The acid condition can irritate the skin, whereas if the pH is too alkaline, it can also dry the skin¹⁸.

The viscosity and the surface tension determination showed the acceptable value for all formula. The emulsion type determination showed a homogenous blue color of methylene blue which means both formulas were oil in water (o/w) emulsion type and all formula were homogenous.

Table 3. The Physico-chemical Evaluation for Nanoemulgel

Formula	pH	Viscosity (cps)	Surface tension (N/m)
F1	5.38 ± 0.20	31.3 ± 0.50	40.63 ± 0.20
F2	5.77 ± 0.23	20.0 ± 0.27	40.50 ± 0.30

Fig. 1: Particle Size Distribution of F1 (A) and F2 (B)

The average globule size for the F1 and F2 were 12.6 and 42.0nm, respectively. The graphic of particle size distribution for F1 and F2 can be seen in Fig.1. All of the nanoemulgel produced showed particle size distribution below than 50nm. The small particle size is expected to increase the nanoemulgel stability for long time storage duration.

The stability of the nanoemulgel was evaluated by storing it at room temperature for 1 month and showed no separation phase and no changes in pH and viscosity of the nanoemulgel. All of the formula showed very good stability. The pH value and the viscosity value for 1 month can be seen from Fig. 2 and 3, respectively.

Fig. 2: pH of Nanoemulgel for 1 Month at Room Temperature

Fig. 3: Viscosity of Nanoemulgel for 1 Month at Room Temperature

Fig.4: pH of Nanoemulgel after Accelerated Stability Test

Fig. 5: Viscosity of Nanoemulgel after Accelerated Stability Test

The pH and the viscosity data after accelerated stability test can be seen in Fig. 4 and 5, respectively. The purpose of the accelerated stability test was to evaluate the stability of the nanoemulgel formulation which it was not kept at room temperature or normal condition. There were no significant changes observed in both formulas after centrifugation, heating cooling cycle and shaking tests. This result demonstrated that the formula of nanoemulgel for both F1 and F2 were stable in any condition.

Stability in pharmaceutical dosage form is one of the important parameters to be considered. A good stability condition of a dosage form will ensure the active ingredients remain stable; therefore, it will give the expected therapeutic effect. Nanoemulsion dosage form gains many advantages such as easily to be removed, stainless, soluble in water and has longer shelf life. Nanoemulsion will become nanoemulgel when incorporating with gelling agent. A stable nanoemulsion preparation will increase the nanoemulgel stability by decreasing the surface and interfacial tension¹⁹.

CONCLUSSION:

The extract of C. mangga can be successfully loaded in nanoemulgel formulation and showed a very good stability on storage at room temperature for 1 month and after accelerated stability test.

ACKNOWLEDGEMENTS:

This research was funded by Universitas Sumatera Utara in accordance with the contract of TALENTA research implementation of Universitas Sumatera Utara for 2019, no. 4167/UN5.1.R/PPM/2019 date 1April 2019.

CONFLICT OF INTEREST:

Authors have no conflict of interest to declare.

REFERENCES:

1. Jambunathan S. Dharani B. Palghat RP. Sumathi S. Cytotoxic activity of the methanolic extract of leaves and rhizomes of Curcuma amada Roxb against breast cancer cell lines. Asian Pacific Journal of Tropical Medicine 2014; 7(Suppl 1): 405-409. doi.org/10.1016/S1995-7645(14)60266-2

2. Kamazeri T. Othman AS. Muhammad T. Deny S. Haitham Q. Antimicrobial activity and essential oils of Curcuma aeruginosa, Curcuma mangga, and Zingiber cassumunar from Malaysia. Asian Pacific Journal of Tropical Medicine 2012; 5(3): 202-209. doi.org/10.1016/S1995-7645(12)60025-X

3. Tedjo A. Dondin S. Latifah KD. Chemoprevention activity of temu mangga extracts. Makara of Health Series 2005; 9(2): 57-62. https://doi/org/10.7454/msk.v9i2.400

4. Pujimulyani D. Wisnu AY. Astuti S. Sela A. Rizal R. Antidiabetic dan antioxidant potentian of Curcuma mangga Val extract and fraction. Asian Journal of Agriculture and Biology 2018; 6(2): 162-168.

5. Muchtaromah B. Alif QAL. Silvia A. Romaidi. Tanjina S. Amaq F. Emy KS. Effect of Allium sativum, Curcuma mangga and Acorus calamus Combination on the Uterus and Hormonal Profile in Rat Induced by Cisplatin. Research Journal of Pharmacy and Technology 2020; 13(11):5438-5442. doi.org/10.5958/0974-360X.2020.00949.X

6. Gutierez JM. Gonzales C. Maestro A. Sole I. Pey CM. Nolla J. Nanoemulsions; new application and optimization of their preparation. Current Opinion & Colloid Interface Science 2008; 13: 245-51. https://doi.org/10.1016/j.cocis.2008.01.005

7. Thakur A. Manpreet KW. Hari K. Nanoemulsion in Enhacement of Bioavaibility of Poorly Soluble Drugs: A Review. Pharmacophore 2013; 4(1): 15-25.

8. Lawrence MJ. Rees GD. Microemulsion-based media as novel drug delivery system. Advanced Drug Delivery Reviews 2000; 45: 89-121. doi.org/10.1016/s0169-409x(00)00103-4

9. Kaur G. Bedi PMS. Jasjeet KN. Topical Nanoemulgel: A Novel Pathway for Investigating Alopecia. Journal of Nanomedicine & Nanotechnology 2017; 8(6): 1-5. doi.org/10.4172/2157-7439.1000472

10. Prasetyo BE. Karsono. Sakro MM. Lia L. Formulation and Physical Evaluation of Castor Oil based Nanoemulsion for Diclofenac Sodium Delivery System. Research Journal of Pharmacy and Technology 2018; 11(9): 3861-3865. doi.org/10.5958/0974-360X.2018.00707.2

11. Alaayedi M. Mahmood H. Saeed A. The Enhancement Effect of Castor Oil on the Permeability of Flurbiprofen as Transdermal Gel. International Journal of Applied Pharmaceutics 2018; 10(1):140-144. Doi.org/10.22159/ijap.2018v10i1.23348

12. Solans C. Isabel S. Nano-emulsions: Formation by low energy methods. Curent Opinion in Colloid & Interface Science 2012; 17(2): 246-254. doi.org/10.1016/j.cocis.2012.07.003

13. Chaudhary H. Rohilla A. Rathee P. Kumar V. Optimization and formulation design of carbopol loaded Piroxicam gel using novel penetration enhancers. International Journal of Biological Macromolecules 2013; 55: 246-253. doi.org/10.1016/j.ijbiomac.2013.01.015

14. Sanjeewani NA. Sakeena MHF. Formulation and Characterization of Virgin Coconut Oil (VCO) Based Emulsion. International Journal of Scientific and Research Publications 2013; 3(12): 1-6.

15. Khan MW. Ahsan MJ. Gupta SK. Development and Invivo Characteerization of Mucoadhesive Nanoemulgel (MNEG) for Enhanced Delivery of Carbamazepine. International Journal Of Advances In Pharmacy Medicine and Bioallied Sciences 2017; 1-9.

16. Safitri FI. Desy N. Dina F. Overview: Application of Carbopol 940 in Gel. Proceedings of the International Conference on Health and Medical Sciences. Advances in Health Sciences Research 2021;34 : 80-84. doi.org/10.2991/ahsr.k.210127.018

17. Varges RP. Costa CM. Fonseca BS. Naccache MF. Mendes PRD. Rheological characterization of carbopol ® dispersions in water and in water/glycerol solutions. Fluids 2019; 4(3). doi.org/10.3390/fluids4010003

18. Yani TN. Effionora A. Fadlina CS. Emulgel Formulation of Binahong Leaves Ethanolic Extract (Anredera cordifolia (Ten.) Steenis) and In Vitro Activity Test Against Propionibacterium acnes secara in Vitro. The Indonesian Pharmaceutical Journal 2016; 6 (2): 89-97. doi.org/10.22435/jki.v6i2.2923

19. Chellapa A. Aref TM. Eseldin IK. Assad E. Ahmad ME.Yosef SI. Nagib AE. Nanoemulsion and Nanoemulgel as a Topical Formulation. IOSR Journal Of Pharmacy 2015; 5(10): 43-47.

Received on 27.04.2021 Modified on 11.10.2021

Research J. Pharm. and Tech. 2022; 15(7):3020-3024.

DOI: 10.52711/0974-360X.2022.00504

Parameters	F1		F2
Parameters	After production	After 1 month	After production	After 1 month
Consistency	Semisolid	Semisolid	Semisolid	Semisolid
Color	Yellowish	Yellowish	Yellowish	Yellowish
Transparency	Cloudy	Cloudy	Transparent	Transparent
Phase Separation	No separation	No separation	No separation	No separation