Formulation and Characterization of a multiple Emulsion containing Vitamin - C

 

M. Venkata Swamy¹*, Samyuktha Metta², A. Ajay Kumar³, C. Jagan Mohan Reddy⁴,

G. Dharani⁵, K. Ramya⁶

¹Associate Professor, Department of Pharmaceutics,

Marri Laxman Reddy Institute of Pharmacy, Hyderabad, Telangana-500043, India.

²Associate Professor, Department of Pharmaceutics,

Marri Laxman Reddy Institute of Pharmacy, Hyderabad, Telangana-500043, India.

³Department of Pharmaceutics, Marri Laxman Reddy Institute of Pharmacy, Hyderabad, Telangana-500043, India.

⁴Department of Pharmaceutics, Marri Laxman Reddy Institute of Pharmacy, Hyderabad, Telangana-500043, India.

⁵Department of Pharmaceutics, Marri Laxman Reddy Institute of Pharmacy, Hyderabad, Telangana-500043, India.

⁶Department of Pharmaceutics, Marri Laxman Reddy Institute of Pharmacy, Hyderabad, Telangana-500043, India.

 

ABSTRACT:

Multiple Mixes have been proposed to have multitudinous uses including their use for improvement of bioavailability or as a dragged medicine delivery system. But the essential insecurity of this system needs to be overcome before they find implicit operation in medicinals. Multiple mixes are frequently stabilized using a combination of hydrophilic and hydrophobic surfactants. The rate of these surfactants is important in achieving stable multiple mixes. The purpose of the study was to prepare a stable multiple conflation containing a skin anti-aging agent and using paraffin oil emulsion. Vitamin C, was incorporated into the inner waterless phase of water- in- oil emulsion- in- water (w/ o/ w) multiple conflation at a attention of 1. Multiple conflation was prepared by two step system. Stability studies were performed at different accelerated conditions, i.e. 8°C (in refrigerator), 25°C (in roaster), 40°C(in roaster), and 40°C at 75 RH (in stability cabin) for 28 days to prognosticate the stability of phrasings. The ideal of this study was to prepare multiple conflation of Ascorbic acid by two step emulsification using different non-ionic surfactants, Tweens and Agar, and estimate for stability, chance medicine ruse. The study concluded that stable multiple conflation with high ruse effectiveness can be prepared by two step emulsification system using agar- agar primary emulsifier and Tween 80 as secondary emulsifier. F2 formulation is the best formulation than the F1 and F3 formulations based on phase separation and other tests.

 

 

 

 

INTRODUCTION:

Multiple mixes are defined as mixes during which both types of mixes, i.e. water- in- oil painting (w/o) and oil painting- in- water (o/w) live contemporaneously. They combine the parcels of both w o and o/ w mixes. These are described as miscellaneous systems of one immiscible liquid dispersed in another in the form of driblets, These two liquids forming a system are characterized by their low thermodynamic stability. Multiple conflation is new approach on medicine delivery system during which the conflation system can live both types of conflation (water in oil painting (W/O) and oil painting in water (O/W) live contemporaneously. The mixture of both parcels of conflation are known as Multiple conflation. Multiple conflation is substantially divided into two types, first is W/O/W (water in oil painting conflation dispersed in water) and another is O/ W/O (oil painting in water conflation dispersed in oil painting). In W/O/W multiple conflation is polydispersible vesicular system during which the dispersed oil painting drops contain indeed lower dispersed driblets, which generally correspond of liquid just like nonstop phase. The presence of force phases inside the drop system are often used for prolonged release of active constituents, Multiple conflation has the numerous operations similar as used in medicinal diligence and ornamental diligence (2). The foremost common system for the medication of multiple mixes is the two step emulsification system by using mechanical stirrer and High pressure homogenizer (HPH). Also water- in- oil painting- in- water (W/O/W) multiple mixes are conflation systems where small water driblets are en- trapped within larger oil painting driblets that successively are dispersed in a nonstop water phase. due to the presence of a force phase inside driblets of another phase that can be used to protract release of active constituents. Multiple W/ O/ W mixes contain both W/O and O/W simple mixes and bear a minimum of 2 emulsifiers to be present in the system when prepared using the 2- step system, one that features a low Hydrophile- Lipophile Balance (HLB) value to stabilize the primary W/O conflation And bone that has a high HLB value to stabilize the secondary O/W conflation. The low- HLB surfactant is dominantly hydrophobic and is added to the oil painting phase. The high- HLB surfactant is dominantly hydrophilic and is added to the external nonstop waterless phase (4). the eye rate of these two surfactants is important to gain stable and high yields of W/O/W mixes.

 

Advantages of multiple conflation:

·      Cover active medicine from declination. High encapsulation effectiveness.

·      Dragged or controlled medicine release are often achieved.

·      Easy to supply and gauge up.

·      Provident.

 

MATERIALS AND METHODS:

MATERIALS:

Ascorbic acid, sulfate, Agar-Agar, paraffin, water, Tween 80, Triethanolamine, Rose oil

METHODS:

The preparation methods of multiple emulsions are often divided into one-step and two-step procedures. Multiple emulsions of the W/O/W type are generally prepared by a two- step procedure^5,6.

 

Preparation of primary emulsion.

Secondary emulsion.

 

Evaluation tests of the multiple emulsion:

1. Physical analysis 2. Microscopic test 3.Centrifugation test 4.Globule size 5.pH determination 6.Rheological tests 7.Stability Test^7,8.

 

Physical analysis:

Organoleptic characteristics: A drop of multiple emulsion was placed on the glass slide, diluted with water and covered by a glass cover and observed under the microscope. Organoleptic characteristics (i.e., color, liquefaction and phase separation) of both primary and multiple emulsions kept at different storage conditions (8°C, 25°C, 40°C, 40°C/75% RH) were noted at various intervals, i.e. 0 h, 24 h, 1 month^9,10.

 

Microscopic test:

Multiple emulsions were analyzed under the microscope to verify the multiple characters. A drop of multiple emulsion was placed on the glass slide, diluted with water and covered by a glass cover. A drop of immersion oil was placed on the duvet slide and observed under the microscope^11,12.

 

Centrifugation test:

Centrifugation tests were performed for the first and multiple emulsions immediately after preparation. The identical test was repeated for the multiple emulsions after 24h, 3days, 7days, 14days and 28days of preparation. Centrifugation conditions were 25°C and 5000 rpm by placing 10 g of sample within the tube^13,14.

 

Globule size:

Globule sizes of the prepared multiple emulsions were determined using microscope fitted with a digital camera for the freshly prepared emulsions as well as emulsions kept at different conditions^{15, 16}.

 

pH determination:

The pH values of the freshly prepared multiple emulsions and emulsions kept at different conditions were determined by a digital pH-meter. pH measurements were repeated for multiple emulsions after 1h, 24h,1months of preparation. Conductivity tests were similarly performed for all the samples by using digital conductivity meter^17,18.

 

Rheological tests:

Rheological parameters were administered for prepared multiple emulsions at different storage conditions by a programmable cone-plate (Brook-field DV III) rheometer. The cone used had an angle of 1.570, radius of 12mm and a shear rate of 3.80 X N (N=rpm). Approximately 0.15g samples at constant temperature were used for the tests. Tests were repeated 3 times, each containing three values of shear rate. Rheograms were fitted to at least one of the available mathematical models. Shear thinning index (ST), thixotropic index (TI) and flow index (FI) were calculated for every sample^19,20.

 

Stability Tests:

Stability tests were performed on samples kept at 4°C±0.1°C (in refrigerator,), 25°C±0.1°C (in oven, Elektro-Mag), 40°C±0.1°C (in oven, Elektro-Mag) and at 40°C±0.1°C with 60% RH (in cabinet, Elektro-Mag). Physical characteristics of multiple emulsions, ie the characteristics mentioned above were all tested^21,22.

 

PLAN OF WORK:

Aim and Objectives: The Aim and Objectives of study is to organize the Ascorbic acid multiple emulation by two step emulsification using different non-ionic surfactants^23,24.

 

 

 

Method of preparation:

Evaluation tests:

1. Physical analysis 2. Microscopic test 3. Centrifugation test 4. Globule size 5. pH determination 6. Rheological tests 7. Stability Tests.

 

METHODOLOGY:

INGREDIENTS AND EQUIPMENTS USED:

Table 1: Ingredients table   

 

Table 2: Equipments table

Equipments	Equipments
Breakers	Magnetic stirrer
Glass rod	Hot air oven
Measuring cylinder	refrigerator
Weighing balance	Centrifuge machine
Water bath	Stabilizer
pH meter	Humidity chamber
microscope

METHOD OF PREPARATION:

Development of formulation:

Table 3: Formulations table

 

Method of Preparation :

Multiple emulsions were prepared by two step emulsification process:

a)    Preparation of primary emulsion.

b)   Secondary emulsion.

 

Primary emulsion:

For the preparation of primary emulsion, oil phase consisting of paraffin oil (13%) and lipophilic surfactant (agar-agar) (2%) was heated to 75°C±1°C. Aqueous phase consisting of Ascorbic acid (1%) and magnesium sulfate (0.5%) was also heated to the same temperature. Aqueous phase was added to the oil phase drop by drop while stirring at 2000rpm. Agitation was continued until cooling to room temperature of 25°C^25,26.

 

Secondary emulsion:

Primary emulsion (57%) was added to the aqueous phase containing hydrophilic surfactant (Tween® 80) (1.0%) while agitating at 1000rpm for 10 min. Emulsion was then homogenized at 800rpm for 5 min and further at 500rpm for 5 min more.

 

RESULTS AND DISCUSSION:

Calibration curve of pure L- Ascorbic acid

Table 4: Calibration curve of pure L- Ascorbic acid

 

 

Fig 1:Calibration curve of pure L- Ascorbic acid

 

FT-IR studies of L-Ascorbic acid with Excipients:

 

Fig 2: FT-IR studies of L-Ascorbic acid pure and with Excipients

 

Table 5: FT-IR studies table

 

Characterization and Evaluation of multiple emulsion containing vitamin-C.

 

Organoleptic Test:

The organoleptic characteristics for the multiple emulsions kept at different storage conditions are observed. Liquefaction was not observed in any of the multiple emulsions stored at different conditions. There was no color change observed in formulations (F1, F2, F3) stored at 8°C and 25°C after 1month. This was confirmed by the absence of phase separation in these samples of formulations (F2). No phase separation was seen in any of the samples of formulations (F2) kept at different storage conditions up to one month. There was slight color change and phase separation observed in formulation F1 and F3.

 

Table 6: Organoleptic characteristics of freshly prepared multiple emulsion:

 

Table 7: Organoleptic characteristics of multiple emulsion.

 

Centrifugation test:

Results of the centrifugation tests for the multiple emulsions kept at different storage conditions (8⁰C, 25⁰C, 40⁰C) are observed. For the multiple emulsion, no phase separation was seen after centrifugation in F2 kept at different storage conditions up to 1 month, however slight phase separation was determined after 1 month in the formulation (F1 and F3)

 

Table 8: centrifugation test of freshly prepared multiple emulsion:

 

Table 9:  centrifugation test of multiple emulsion:

 

Microscopic test:

Microscopic analysis was carried out using an research microscope and observations were made at 40 X magnification after diluting in the appropriate amount of external phase of the emulsion the image taken via microscope.

 

 

Figure.3: Microscopic image of (a) freshly prepared multiple emulsion, (b) at 8°C (in refrigerator), (c) at 25°C (in oven), (d) at 40°C (in oven).

 

Globule size:

Emulsion globule sizes of the samples of multiple emulsions at different storage conditions and time intervals were observed. Mean globule size of the freshly prepared multiple emulsion formulation was 8.46μm. Mean globule sizes of the multiple emulsion samples kept at 8°C were increased from 8.12μm to 8.32μm after 1 months. Marginal increase in globule size from 8.22μm to 8.52μm was shown by the samples stored at 25°C.

 

Table 10: Globule size of freshly prepared multiple emulsion.

 

Table 11: Globule size of multiple emulations stored at different conditions.

 

pH determination:

pH values of the freshly prepared formulation was 5.84±0.01, 5.86±0.01 (F2, F3) and 5.82±0.05 (F1). The pH values of the formulations kept at different storage conditions have decreased with time. The pH values of the formulations were found to be quite stable when stored at 25°C for 1month. It was slightly changed when stored at 8°C after 1 month. Marked variations of pH were observed, when kept at 40°C.

 

Table 12: pH value of freshly prepared multiple emulations

 

Table 13: pH values of multiple emulations stored at different conditions

 

Stability Tests:

Stability tests were performed on samples kept at 4°C±0.1°C (in refrigerator), 25°C±0.1°C (in oven), 40°C±0.1°C (in oven). Physical characteristics of multiple emulsions, i.e the characteristics mentioned above   were all tested.

 

 

(a)

 

 

(b)

 

 

(C)

Fig 4: (a) stirring of emulsion in beaker side view (b) top view of stirring of emulsion  in beaker (c) front view of stirring of emulsion in beaker

 

SUMMARY AND CONCLUSION:

Multiple emulsions have been proposed to have numerous uses including their use for enhancement of bioavailability or as a prolonged drug delivery system. Multiple Emulsion is prepared by two step emulsification method in which first step under formation of pre- emulsion (W/O) and second step under treatment of aqueous water phase emulsified with previously prepared W/O emulsion to form W/O/W type of multiple emulsion. Using different W/O emulsions for the second emulsification step the finest W/O emulsion led to W/O/W emulsions with the highest encapsulation rate. To prevent a production-induced reduction of encapsulation rate the inner water droplets have to be much smaller than the oil droplets in the Multiple Emulsion. From this investigation it was found that stabilized. Ascorbic acid multiple emulsions could be formulated using magnesium sulfate as stabilizer. There was changes in globule size, color change, phase separation, pH and in conductivity observed in the formulations F1, F3 stored at 40°C. But there was only marginal change in pH, conductivity, globule size, and rheological parameters for multiple emulsions of formulation F2 stored at 25°C.

 

It can be concluded that the multiple emulsion prepared using paraffin oil containing Ascorbic acid as active ingredient is a very stable multiple emulsion system. From the above results F2 formulation found as the best formulation.

 

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Received on 27.06.2022       Modified on 14.08.2022

Accepted on 28.10.2022   ©AandV Publications All Right Reserved