INTRODUCTION:

Alzheimer’s disease, named after the doctor Alois Alzheimer^{1}. This somatic disease affects the brain⁽²⁾. it is a progressive disease that destroys memory and other important mental functions⁽³⁾. During this disease, protein builds up in the brain and form structures called ‘plaques’ and ‘tangles’^(3,4). This results in the loss of connection between nerve cell⁽⁵⁾, and lead to the degeneration of neurons^(5,6) and finally death of brain tissue⁽⁷⁾.

Due to deficiency of some important chemical messenger in the brain, the signal transmission gets ultimately stopped⁽⁸⁾, and signals are not transmitted as effectively in the body^(7,8).

As several studies have reported the deficiency of cholesterol in brain of AD patients⁽⁹⁾the Current therapy of virgin coconut oil emulsion for treatment of Alzheimer’s disease may help to increase the cholesterol level in the brain and to destroy the β amyloid plaque which ultimately enhances the level of chemical messengers in the brain.

Coconut (Cocosnucifera, family Arecaceae ⁽¹⁰⁾) oil emulsion consist medium chain triglycerides (MCTs)^(10,11). These MCTs are broken down by enzymes in the saliva and gastric juices⁽¹²⁾ and converts into ketone bodies^(12,13), these ketone bodies enters into the brain by crossing blood brain barrier⁽¹⁴⁾ and results into the destruction of β amyloid plaque⁽¹⁵⁾ which eventually improves the signal transmission in the brain⁽¹⁶⁾.

An emulsion is a thermodynamically unstable system⁽¹⁷⁾. It consists of two phases (oil phase, aqueous phase), one of which dispersed as droplet into another^(17.18). The stability of emulsion plays a significant role in ensuring its quality⁽¹⁹⁾. The stability factor may sometimes get affected by the physicochemical property of the gums added during the preparation of emulsion⁽²⁰⁾.coconut oil appears clear (like water), with a coconut flavor and sweet aroma and no rotten smell, even without undergoing any chemical refining processes^(19,20). In the late 1990s the clear version of coconut oil (VCO) became popular in the market because of its outstanding characteristic of containing high content of medium chain fatty acids (MCFAs)⁽²¹⁾Future prospective of this research is to perform in vivo studies for reducing the dose of coconut oil emulsion and to make its targeted drug delivery system.

MATERIALS AND METHOD:

Materials:

Virgin coconut oil is extracted in the laboratory ofRungta College of Pharmaceutical Science and Research. Distilled water and Tween 20 is also taken from pharmaceutical laboratory of RCPSR

Methods:

1. Method of extraction of virgin coconut oil^(22.23)

2. Preparation of emulsion^(24,25,27)

The emulsion was formulated in 1MLH magnetic stirrer in laboratory of department of pharmacy Rungta College of Pharmaceutical Sciences and Research kohka, kurud road Bhilai Chhattisgarh.

In this study emulsion of different ratio were prepared.

Four formulations were prepared in different ratios of oil, water and surfactant for the preliminary studies.

Table 1: Formulation table

S.NO.	Sample name	Oil (g)	Water (g)	Surfactant (g)	Oil : Water: Surfactant (% w/w)
1	P	5.1	4.8	5.1	34:32:34
2	Q	4.8	5.4	4.8	32:36:32
3	R	5.4	4.2	5.4	36:28:36
4	S	5.7	3.6	5.7	38:25:38

CHARACTERIZATION:

After getting the best formulae based on accurate oil, water and surfactant ratio, it was further studied for its characterization such as pH value, creaming index, viscosity and microscopic studies. All this studies are conducted in the laboratory of RCPSR.

Physical appearance:

The physicalappearance test of Coconut oil emulsion is done by observing it through sensory organ for determine its color, odour, taste appearance.^{(26, 27, 28)}

Dilution test:

in emulsion, if continuous phase is added or mixed, the emulsion will not separate or crack into phases^{(29, 30)}. Example, if continue phase like water is added in to the O/W type of emulsion. It will remain stable.

Creaming:

Creaming may be defined as formation of a layers of dispersed phase in relatively concentrated form at the surface^(31,32). Such an emulsion on vigorous shaking results in homogeneous emulsion. On standing again, the cream forms on the surface. It is undesirable because this leads to cracking of emulsion resulting in an inaccurate dose.

20 grams of sample of coconut emulsion were transferred into a bottle and sealed with cap and stored for 24 h at 25⁰C. In general, dispersed phase has a lower density than the continuous phase. So that the oil phase move upward which leads to creaming during storage^(33,
34).

Creaming index = 100 (HD/HE)

pH value:

pH values were measured at room temperature by using a digital pH meter (modal no. 111 ) . Three readings were taken for each sample. Before recording the readings, pH meter was calibrated by using buffer solutions of pH 7.4 and 10 respectively⁽³⁵⁾.

Dyes test / microscopic test:

1. METHYLENE BLUE TEST – It is a water soluble dye used to determine the type of emulsion (W/O or O/W) methylene blue is mixed with one drop of emulsion in the slide and spreaded with the help of spatula, covered with cover slip and microscopical evaluation is performed. Appearance of continuous blue phase indicates that emulsion is oil in water type and white outer phase with dark bluish globules indicate the water-in-oil type.^(36,37)

2. SUDAN III TEST – It is an oil soluble dye used to determine the type of emulsion (W/O or O/W) sudan III is mixed with one drop of coconut oil emulsion on the slide and spreaded with the help of spatula, covered with cover slip and microscopical evaluation is performed. Appearance of dark colored globule indicates that the emulsion is oil in water type and colorless outer phase indicate the water-in-oil type emulsion ⁽³⁸⁾.

3. AMARANTH DYE TEST – it is a water soluble dye used to identify the type of emulsion (whether it is W/O or O/W), Amaranth dye added on the sample of coconut oil emulsion and evaluated in the microscope, the appearance of colored dispersion medium that indicates the emulsion is O/W type, and appearance colorless globules which indicate the emulsion is W/O type.^{(39, 40)}

Viscosity measurement:

Viscosity of the selected formulation was measured by using the Brookfield viscometer at room temperature. This viscometer is used for determining the viscosity of oils. The value of viscosity is expressed in centipoise. Centipoise is defunct unit of viscosity measurement^{(41, 42)}. Firstly the Spindle number of Brookfield viscometer was found out by trial and error method; after that sample was poured in the beaker and spindle was dipped into it to find out the viscosity at different RPM. The value of torque and viscosity was then recorded ^{(43, 44)}.

Stability:

Approximately 50 ml of sample was put into a bottle and stored at 14-21 ⁰C, 21-25 ⁰C and 25-30 ⁰C for the period of 28 days. The stability of VCOE was recorded.⁽⁴⁵⁾

RESULT AND DISCUSSION:

1. Physical appearance - the physicalappearance test of l emulsion is done by observing it through sensory organ and following observation is made.

Table 2: Organoleptic properties

Color	Milky white
Odour	Sweet, aromatic
Taste	Sweet
Appearance	Viscous

2. Creaming:

Samples didn’t showed creaming property, after storing it for 24 hours. Therefore samples have zero creaming indices.

3. pH value:

pH values of the sample is measured by using pH meter of model number 111.The graph indicates that all the resulted pH values are in range between5.76– 7.88. These values indicate that emulsion is suitable for oral administration. .

Table 3: pH value of Formulation

SNO	SAMPLE	pH
1	P	5.76
2	Q	6.78
3	R	7.74
4	S	7.89

Fig 1: graphical representation of pH values

DILUTION TEST

When oil is added in the samples (P,Q,R,S) the emulsion get separated into two phases.

VISCOSITY MEASUREMENT:

Table 5: Result recorded for viscosity measurement

SNO	SAMPLE	VISCOSITY {Cp}
1	P	525
2	Q	527
3	R	523
4	S	524

Fig 3: Brook field viscometer model-- AVDV

Fig 4: graphical representation of viscosity

STABILITY TEST

After 7 days

Table 6: Stability data after 7 days

SNOo	SAMPLE	TEMPERATURE (⁰C)
SNOo	SAMPLE	14-21	21-25	25-30
1	P	Stable	Stable	Stable
2	Q	Stable	Stable	Stable
2	R	Stable	Not stable	Not stable
4	S	Stable	Stable	Not stable

After 14 days

Table 7: Stability data after 14 days

SNO	SAMPLE	TEMPERATURE (⁰C)
SNO	SAMPLE	14-21	21-25	25-30
1	P	Stable	Stable	Stable
2	Q	Stable	Stable	Stable
2	R	Stable	Not stable	Not stable
4	S	Stable	Stable	Not stable

After 21 days

Table 8: Stability data after 21 days

SNO	SAMPLE	TEMPERATURE (⁰C)
SNO	SAMPLE	14-21	21-25	25-30
1	P	Stable	Stable	Not stable
2	Q	Stable	Stable	Stable
2	R	Not stable	Not stable	Not stable
4	S	Stable	Not stable	Not stable

After 28 days

Table 9: Stability data after 28 days

SNO	SAMPLE	TEMPERATURE (⁰C)
SNO	SAMPLE	14-21	21-25	25-30
1	P	Stable	Not stable	Not stable
2	Q	Stable	Stable	Not Stable
2	R	Not stable	Not stable	Not stable
4	S	Not stable	Not stable	Not stable

DISCUSSION:

The important criterion for selection of components for emulsion formulation is their compatibility with other component. It has been demonstrated that only few excipients combinations lead to effective emulsion formulations

VCO was selected as oil phase for the development of the emulsionbecause it is known to improve the symptoms related with Alzheimer’s disease. In the year 2003, Indian biochemists investigated the effect of VCOE on various lipid parameters in albino rats. VCO has also its effect in lowering lipid component compared to copra oil and ground nut oil. It reduces total cholesterol, phospholipids, LDL and VLDL levels in the body and help to increase HDL cholesterol in blood serum and tissues.

Fatty acid (MCFA) can act as a non-carbohydrate source by enhancing the formation of ketones or ketone bodies. Fatty acids itself cannot pass the blood–brain barrier (BBB); thus, the human brain primarily depends on glucose. However, it can utilize alternative fuels such as ketones to maintain energy and ketone bodies are used extensively as an energy source during glucose deficiency. These short-chain organic compound (ketone bodies) can penetrate cell membranes liberally and BBB through proton-linked, monocarboxylic acid transporters.

Maintaining the pH value is significant for determining the stability of the emulsion because change in pH specifies the occurrence of chemical reactions in the. The resulting pH of sample P, Q,R,S is found as 525, 527,523,524 respectively. The sample Q with grater viscosity of 527 is found to be more stable.

ACKNOWLEDGEMENT:

Authors want to acknowledge the facilities provided by the Rungta College of Pharmaceutical sciences And Reseach, kohka, Kurud road Bhilai, Chhattisgarh. The authors also grateful to MrGyaneshSahu Sir for his help to compete work in stipulated period of time.

CONCLUSION:

This study is further aimed to perform in vivo studies for analyzing the concentration of ketone bodies reaching into the brain and to study its effect in removing β amyloid plaque, which will help to reduce the dose of coconut oil emulsion and to make its novel dosage form for treatment of Alzheimer’s disease.

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Received on 10.04.2018 Modified on 11.05.2018

Res. J. Pharma. Dosage Forms and Tech.2018; 10(2): 49-54.

DOI: 10.5958/0975-4377.2018.00009.5

SNO	TEST	TYPE OF DYE	RESULT	INFERENCE
1	METHYLENE BLUE TEST	Water soluble		Continuous blue phase indicate that emulsion is oil in water type (O/W)
2	SUDAN III TEST	Oil soluble		Appearance dark brown color globule indicate that emulsion is oil in water type (O/W)
3	AMARANTH TEST	Water soluble		Appearance Continuous red phase indicate that emulsion is oil in water type (O/W)