Sunday, 12 May 2013

Emulsion Part II


Discussion
1.  What are the HLB values that will produce a stable emulsion? Discuss.

  
Eight simple emulsions were prepared in eight test tubes each with the same amount of oil and distilled water which is 4mL, surfactant but with a different HLB values. The test is conducted with different types of oil which are Palm Oil, Arachis Oil, Olive Oil, and Mineral Oil. The emulsion appears to be the most stable were observed. Stable emulsion meaning here is the one least separated or the one that separates last. The HLB value of the surfactant used in this emulsion is the HLB requirement for that particular oil phase. The HLB of an emulsifier is an expression of its Hydrophile-Lipophile Balance, which are the balance of the size and strength of the hydrophilic (water-loving or polar) and the lipophilic (oil loving or non-polar) groups of the emulsifier. All emulsifiers consist of a molecule that combines both hydrophilic and lipophilic groups. An emulsifier that is lipophilic in character is assigned a low HLB number (below 9.0), and one that is hydrophilic is assigned a high HLB number (above 11.0). Those in the range of 9-11 are intermediate. When two or more emulsifiers are blended, the resulting HLB of the blend is easily calculated by using the formula:
HLB value =
(quantity of surfactant 1)(HLB of surfactant 1)+(quantity of surfactant 2)(HLB of surfactant 2)
quantity of surfactant 1 + quantity of surfactant 2

The HLB of an emulsifier is related to its solubility. Thus, an emulsifier having a low HLB will tend to be oil-soluble, and one having a high HLB will tend to be water-soluble, although two emulsifiers may have the same HLB and yet exhibit quite different solubility characteristics. The SPAN emulsifiers are usually lipophilic and the TWEEN products are usually hydrophilic.



2. Compare the physical appearance of the mineral oil emulsions produced and give your comments. What is Sudan III test? Compare the colour dispersion in the emulsions produced and give your comments.

In this experiment, Emulsion I contains 20ml of mineral oil, Emulsion II contains 25 ml of mineral oil, Emulsion III contains 30ml of mineral oil while Emulsion IV contains 35ml of mineral oil.
Before homogenization, generally Emulsions Ι, ΙΙ, ΙΙΙ and ΙV are less viscous, more oily and not consistent. Phase inversion is likely to occur. After homogenization, generally Emulsions Ι, ΙΙ, ΙΙΙ and ΙV are more viscous, less oily and more consistent. Emulsions produced after homogenization is more stable.
Sudan III stain is a biological stain used to stain sudanophilic substances, usually lipids and it is usually provided as a saturated alcoholic solution. Sudan dyes have high affinity to fats; therefore they are used to demonstrate triglycerides, lipids, and lipoproteins. Thus, Sudan III test can be used to differentiate between oil-in-water emulsion and water-in-oil emulsion. If Sudan III solution is added to an emulsion and the disperse phase appears red, then it is oil-in-water emulsion. However this test is only applicable if the non-ionic emulsifier is used in the preparation (e.g. Tween 80 and Span 20).
The red stain of Emulsions Ι, ΙΙ, ΙΙΙ and ΙV is uneven before homogenization. So, we can say that these emulsions formed are water-in-oil emulsion. However, after homogenization, the red stain is even. Red globules have been seen in uniform dispersion on a colourless background. The size of the globules is small. Hence, oil in water emulsion is formed after homogenization. Here, phase inversion occurred.


3. Plot and give comments on:

a) Graph of the sample’s viscosity before and after the temperature cycle against the different amount of oil content.

Amount of Oil (ml)
Average Viscosity (cP) (x ± SD)
Viscosity difference (%)
(x ± SD)
Before temperature cycle
After temperature cycle
20 mL of Palm oil
78.33 ± 57.57
114.17 ± 103.86
45.76% ± 80.41%
25 mL of Arachis oil
60.00 ± 28.28
105.00 ± 11.18
75% ± 60.47%
30 mL of Olive oil
490.00 ± 46.19
2626.67 ± 219.97
436.06% ± 376.23%
35 mL of Mineral oil
7363.33 ± 1563.83
11325.00 ± 4129.14
53.80% ± 164.04%






In the experiment, we use different oils to prepare 4 types of emulsion but the same emulsifying agent is used for both emulsions. The emulsifying agent used is acacia. Acacia is a hydrophilic colloid that forms multimolecular adsorption at the oil/ water interface. It has low effect on the surface tension. Its main function as emulsion stabilizers is by making coherent multi-molecular film. This film is strong and resists the coalescence. They have, also, an auxiliary effect by increasing the viscosity of dispersion medium. The HLB value of acacia is 8 and form oil-in-water emulsions. Acacia contains Arabic acid and proteins (COOH and NH3) and provides electrostatic repulsion.
  Different formulations of emulsion have different physical properties and also stability. Viscosity is one of the physical properties that used to characterize emulsion. Temperature cycle is one of the accelerated stability testing method. In the experiment, different amount of oils (20mL of palm oil, 25mL of arachis oil, 20mL of olive oil and 35mL of mineral oil) are used in the formulation of emulsion. Oil has higher viscosity than water. Thus, emulsion with higher amount of oil will have higher viscosity. This can be shown on the graph above. From the graph, the viscosity of emulsion increases as the proportion of oil increases. This trend is same for both before and after temperature cycle graphs.
Increase in temperature of emulsion will cause decrease in viscosity of continuous phase and will increase the kinetic motion of dispersed phase. The rate of creaming increases as the viscosity of continuous phase decreases. As the kinetic energy of dispersed phase increases, the globules gain energy and collide with each other more frequently, thus results in breakdown of emulsifying agent and cause coalescence. Partial coalescence could also have occurred during the cooling of the emulsions. At low temperature, cold denaturation of proteins may occurred which could lead to a loss of protein functionality when the emulsions were thawed. Besides, the crystallization of some of the water causes the globules to be forced closer together, thereby promoting globule-globule interactions. When the water phase froze, insufficient free liquid water present to fully hydrate the emulsifying agents present at the globule surfaces, thereby promoting globule-globule interactions. The temperature cycle will disrupt the adsorbed layer of emulsifying agent at the oil/water interphase.
Furthermore, an oil-in-water emulsion stabilized by non-ionic emulsifying agent such as acacia will cause phase inversion to water-in-oil emulsion on heating. This is because as the temperature increases, the HLB value of a non-ionic emulsifying agent will decrease as it becomes more hydrophobic. At the phase inversion temperature (temperature at which the emulsifier has equal hydrophilic and hydrophobic tendencies), the emulsion will be inverted. From the graph, the viscosity of all the emulsions after the temperature cycle increases because the oil-in-water emulsion has become water-in-oil emulsion where oil is the continuous phase, contribute to the higher viscosity of emulsion.


b) Graph of the difference of viscosity (%) against the different amount of oil content. 


   
From the graph above, the viscosity differences before and after the temperature cycle are almost the same which are 45.76%, 75% and 53.8% for emulsions prepared from 20 mL of palm oil, 25 mL of arachis oil and 35 mL of mineral oil respectively. However, there is a very large viscosity difference for emulsion prepared from 30 mL of olive oil which is 436.06%. By theory, larger viscosity difference indicates a weaker and less stable emulsion. As the volume of dispersed phase increases, the stability of the emulsion decreases and phase inversion may occur. The small viscosity differences for emulsions composed of 20 mL of palm oil and 25 mL of arachis may be caused by coalescence of dispersed phase after the temperature cycle, not due to phase inversion. On the other side, the emulsion prepared from 30 mL of olive oil may undergo phase inversion after the temperature cycle. This causes oil phase become the continuous phase which has a much higher viscosity than water, so contribute to much higher viscosity difference.  The emulsion prepared from 35 mL of mineral oil may already undergo phase inversion before the temperature cycle, so  there is only a slight increase in viscosity (small viscosity difference cycle) after the temperature cycle. To make the result more accurate, there are several precaution steps that can be carried out in this experiment. Firstly, stir the emulsion thoroughly before getting the viscosity readings from the viscometer. Secondly, only take readings after the readings in the viscometer become stable and constant. Finally, clean the rotor in the viscometer by dipping it in the distilled water before taking the second and third readings.

4. Plot the graph of the separated phase ratio after centrifugation process against the different contents of mineral oil. Give your comments. 

Ratio of phase separation for different amount of different types of oil

Types of oil (mL)
Ratio of Phase Separation (x ± SD)
Coconut Oil (20mL)
0.715±0.085
Arachis Oil (25mL)
0.670 ± 0.03
Olive Oil (30mL)
0.250 ± 0.03
Mineral Oil (35mL)
0.245 ± 0.025


 
Types of oil
Ratio of Phase Separation
Coconut oil
0.715
Arachis oil
0.670
Olive oil
0.250
Mineral oil
0.245
   


From the graph, it could be seen that the separation ratio decreases from coconut oil to arachis oil to olive oil and to mineral oil. So, as a conclusion, mineral oil has the lowest ratio of phase separation. This may be caused by the structure of mineral oil which is made up of only carbon and hydrogen. This causes the structure to become non polar. When comparing mineral oil and the other oils such as coconut oil, arachis oil and olive oil, they contain carboxylic group which renders them more polar than mineral oil. Hence, if the polarity of an oil is low, it will cause the ratio of phase separation to be low. This will also cause the emulsion formed to be more stable. As such, the separated phase ratio must be kept at minimum to produce a stable and homogenous emulsion to ensure that the drug can be dispersed uniformly in the emulsion and an accurate dose of drug can be taken for each volume of emulsion administered.


5. What are the functions of every substance used in this emulsion preparation? How the different contents of substances can affect the physical characteristics and stability in the formulation of an emulsion?
There are some ingredients that has to be used to prepare an emulsion. They are oil phase (mineral oil for example turpentin oil), aqueous phase (syrup, alcohol and distilled water), emulsifying agent (acacia) and other substances such as vanillin.
First and foremost, the oil phase of the emulsion, given by the mineral oil, functions as the oil phase in this oil in water (o/w) emulsion. Mineral oil is the dispersed phase in this emulsion. The oil content is set in range of 40% to 60% in order to produce a stable o/w emulsion. Besides that, the oil phase of an emulsion can provide emollient effects.
Syrup is responsible as a sweetening agent. This is because emulsion normally will give a taste of oil. To mask the taste of the oil phase, sweetening agent is required for this purpose. Moreover, syrup increases the viscosity of the emulsion. However, the amount of syrup used has to be controlled as this will determine the flow of the emulsion. Sufficient amount of syrup will produce an emulsion which has good flow and can be easily poured out, Besides that, emulsion will not form a viscous layer on the side of the container.
Alcohol functions as a preservative to prevent the growth of this microorganism in the emulsion. Basically, the amount of alcohol used as a preservative depends on the amount of water phase in the emulsion as water is the best medium for microorganisms to grow. Distilled water on the other hand, act as the aqueous phase (external phase or continue phase) in the oil in water (o/w) emulsion.
The next ingredient which is also very important is Vanillin. Vanillin acts as flavouring agent to improve the taste of the emulsion so that it is more acceptable to the patients.
On top of that, another ingredient that is important too is the emulsifying agent. It can increase the viscosity of the surface of the oil and aqueous phase. Besides, it also provides a protective sheath for the oil droplets in emulsion. Acacia is also a natural product and it is a suitable medium for the growth of microorganisms. Hence, antimicrobial agent needs to be added to the emulsion to stabilize it. Acacia will not decrease the surface tension.
The ingredients used in preparing emulsion will affect the physical and chemical characteristics of the emulsion. Different types of mineral oil used in the preparation of emulsion will give rise to a different physical characteristics and chemical stability in an emulsion formulation. Besides that, the different compositions of mineral oil and distilled water used in the preparation of emulsion will affect the types of emulsion produced, either oil in water (o/w) emulsion or water in oil (w/o) emulsion. If the amount of distilled water used is in excess compared to the oil used, then a oil in water emulsion will be produced. While water in oil emulsion is produced if the amount of oil used is in excess. Hence, any change in the content of water or oil will results in phase inversion.
Furthermore, the content of syrup can affect the flow and physical characteristics of the emulsion formed. The content of the syrup used must be controlled to  prevent any problems related to the emulsion's rheology properties. Physical characteristics can also be affected in this case. Some of the oils used have different colours and this will produce emulsion with different colours. In addition, palm oil has antioxidant property and this can improve the stability of the emulsion formed. The differences discussed above will produce an emulsion with different physical characteristics and chemical stability.


Conclusion 
An emulsifier having a low HLB will tend to be oil-soluble emulsion, while a high HLB will tend to be water-soluble emulsion. The less polar of the emulsifier been used, the more stable the emulsion to be formed.

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