1. INTRODUCTION:

Emulsions may well be presented as heterogeneous systems, in which immiscible liquid has been distributed mostly in the form of droplets as well as remained stable by the third component named as emulsifying agent. Those other two substances also are chemically inert and create the systems which are described by even a reduced thermodynamically reliability.

Premised on their development, emulsions can be categorized into:

a) Simple Emulsion-

Simple Emulsions can also be isolated thus according of their continuous medium or dispersion medium as,

a. Oil-in - water emulsions (O / W) – in which oil will be the disperse medium in a continuous phase of water, as well as,

b. Oil-in - water emulsions (O / W) – in which oil will be the disperse medium in a continuous phase of water, as well as, Water-in - oil emulsions (W / O) – in which water will be the disperse phase in a continuous phase of oil.

b) Multiple Emulsions-

Multiple emulsions are indeed multidimensional systems, considered as "emulsions of emulsions", that is the droplets of a dispersion medium includes that much tiny dispersed particles itself. All dissipated globule with the double emulsified aspects a Vesicular framework with single or multiple aqueous compartments segregated from the aqueous medium by a layer with oil medium compartments.1,2,3 Multiple emulsions are mostly known as emulsions of emulsions, liquid phase system or even double emulsion. ^{10, 14, 20}

1.1. Types of multiple emulsions:

1.1.1. Oil in water in oil (o / w / o) emulsion- In O / W / O systems, an aqueous phase differentiates internally and externally oil phases. In many other words, O / W / O is a system in which water droplets may well be covered in an oil phase, which in turn envelops one or more oil droplets.

1.1.2. Water in oil in water (w / o / w) emulsion- In W / O / W systems, an organic phase differentiates internal and external aqueous phases. In several other words saying, W / O / W is a system for which an oil droplet may well be enclosed by such an aqueous phase, which then in turn is encloses yet another or more water droplets. Such systems are far more studied amongst these multiple emulsions. ⁹

1.2. Advantages: ^{17, 18, 21}

1. Masks bitter taste as well as bad smell of medications, while also trying to make them more accessible.

2. A phenomenal degree of bio-compatibility.

3. Enhances discharge of drug, while also supplying sustained release action. Complete biodegradability and the lack of toxic products resulting from carrier degradation.

4. Essential nutrients like carbohydrates, fats and vitamins can all be emulsified.

5. Food items like carbohydrates, fats as well as vitamin supplements all can be emulsifying agent.

6. Avoidances of the any unwanted immune responses against encapsulated drug.

7. Could be administrated with bed ridden patients when sterilized injectable emulsions.

8. Safety of its loaded substance from inactivation by an endogenous variable.

9. Improvement of enteric or dermal uptake.

10. Hydrophilic and also hydrophobic drugs molecules can be entrapped.

11. Improves drug release and therefore improve in drug dosing increments.

1.3. Disadvantages: ¹⁸

1. Thermodynamically unbalanced, possess complicated pattern that also contributes to short lifespan of a product.

2. These are marketed in such a plastic/glass jar container, as such care needs to be taken in transport and storage.

3. Non homogeneous structure.

4. Might cause disruption in substance.

5. Requires vigorous washing to remove the traces of organic solvents.

6. Particulate matter is difficult to see prohibiting visual inspection of solution.

7. Support the growth of microorganisms better than glucose-amino acid solution.

8. Filtration must be performed with a larger filter.

1.4 Limitations of multiple emulsions ^1,
22

The primary issue related to multiple multiple emulsions seems to be their thermodynamic destabilization as well as their complicated system that has severely limited their effectiveness in many of the implementations of multiple emulsions

1. Convergence of multiple emulsion droplets or internal droplets.

2. Fracture of oil layer on exterior of inner drops.

3. Shrinkage as well as swelling of inner droplets due to the osmotic gradient across the oil membrane.

4. Coagulation of inner aqueous medium as well as multiple emulsion droplets and medium separation. The primary issue through specific regard to stabilization seems to be the existence of two interfaces that are thermodynamically unstable. So, 2 different emulsifiers are essential for their stabilization; with a low HLB (Hydrophile-Lipophile Balance) value for W / O interface or the one with a high HLB value for O / W interface. We could also stabilize the emulsions through using electrolytes, whilst creating polymeric film, through interfacial chelating in both non-ionic surfactant and macro molecules.

1.5 Composition of emulsion:¹²

1.5.1 Emulsifying agent

The emulsifying agents reduce the surface tension of two phases i.e., oily phase as well as aqueous phase or even start making others to miscible with one another and create a continuous emulsion. Emulsifying substances are often identified as emulgents or emulsifiers.

Classification of emulsifying agent-

a) Natural -e.g.-Acacia, Tragacanth, Pectin, Starch, Irish mass, Wool fat, Egg yolk, Gelatin.

b) Semisynthetic polysaccharides-e.g.–Methyl cellulose, Sodium carboxymethyl cellulose.

c) Synthetic-e.g.-Anionic, Cationic, Non ionic.

d) Inorganic-e.g.-Milk of magnesium, Magnesium oxide, Magnesium aluminum silicate.

e) Alcohols-e.g.- Carbowax, Cholesterol, Lecithin.

1.5.2 Preservatives

Emulsions that are formulated through using emulsifying agent, like carbohydrate, proteins, sterol and surfactants may lead to the development of bacteria, fungi and moulds in the presence of water. So the preservation is require for that reason preservatives are used.

e.g. - Methyl paraben, Propylparaben, Benzoic acid, Chlorocresol, Chloroform, Cetrimide.

1.5.3 Antioxidants

During storage of emulsion, a fats but also emulsifying agents undergo oxidation whilst atmospheric oxygen. It can be avoided through using antioxidants.

e.g.- Tocopherol, Gallic acid, Propyl gallate, Ascorbic acid.

1.5.4 Flavors

Vanillin is now a good flavouring agent for liquid paraffin emulsions. Benzaldehyde has been generally used as a flavouring agent for cod-liver oil emulsion. A combined effect of flavouring as well as sweetening agent allows additional palatability to emulsion.

2. METHOD OF PREPARATION ¹

Multiple emulsions were prepared by two step emulsification process:

2.1. Preparation of primary emulsification

Main emulsification: 10 ml distilled water usually contains 25 mg of medication has been steadily introduced to 14 ml of oil medium containing primary emulsifying agent (Span 40, Span 60, and Span 80) and 25 mg of medication with constant agitation at 5000 rpm for 5 min. It provides the primary emulsion.

2.2. Preparation Secondary emulsification

Secondary emulsification can be 20 ml of viscous primary emulsion is now emulsified even farther with such an exterior aqueous solution containing secondary emulsifying agent (Tween 80) and 50 mg drug with constant stirring at 1000 rpm for 10 min. All of the formulations were developed whilst following a same method. Impact of primary emulsifier has been noticed by evaluating many other products.

2.3. Behavior of Multiple Emulsions in Biological System: ^{7,
8, 9}

ME's were administered by oral, parentral (i.v., i.m., s.c.) as well as topical routes (nasal, ocular, transdermal) routes. After oral administration ME is almost absorbed entirely from lymphatic pathway in association with intestinal lipoproteins namely chylomicrons, produced by enterocytes. They may directly be absorbed through intestinal macrophage system and Payers Patches to gain access into mesenteric lymph from where they are drained into circulation through thoracic lymph duct. Consequently, they are capable of carrying bioactive components inside of them trying to avoid deterioration throughout intestine as well as liver. After parenteral (i. v. or i. m.) administration the emulsions will be readily absorbed by circulatory macrophage system to lymphatics and liver into fat metabolism pathway through other parenteral routes, the emulsion droplets gain access to nearby lymphatic node through interstitial spaces of lymphatic vessels which are relatively porous as compared to blood capillaries which have tight intracellular junctions.

2.4. Possible mechanism of drug release from multiple emulsions

In multiple emulsions, the drug release from internal to external medium through the oily interface by different method. The release rates were also affected by various aspects like particle sizes, pH, phase volume as well as viscosity etc.

3. THE VARIOUS MECHANISMS ARE: ¹⁰

3.1. Diffusion mechanism

It is most prevalent transport mechanism in which unionised hydrophobic medication disperses through the oil layer mostly in stable multiple emulsions. Medication transport has also been found to follow first order kinetics as well as obeyed Flick’s law of diffusion.

3.2. Micellar transport

Inverse micelles consisting of non polar part of surfactant lying outside and polar part inside encapsulate hydrophilic drug in core and permeate through the oil membrane because of the outer lipophilic nature. Inverse micelle could also encompass respectively ionised and unionised substances. Recently, the release of tetra decane from a tetrad cane/ water/ hexadecane multiple emulsions was investigated using the differential scanning calorimetric technique. Micellar diffusion instead of molecular diffusion was considered to become the preponderant method for mass transfer.

3.3. Hinning of the oil membrane

Due to the osmotic pressure gradient, the oil membrane have become thin, therefore the water as well as drug rapidly diffused. This differential pressure as well does provide force for transverse of molecule.

3.4. Rupture of oil phase

As per this technique rupturing of oil membrane can unite both aqueous medium and thus medication can be released easily.

3.5. Facilitated diffusion (Carrier mediated transport)

This mechanism includes a specific molecule (carrier) that also combines with the drug and makes it compatible to permeate via the oil membrane. Such carriers can also be implemented in inner aqueous phase and oil membrane.

3.6. Photo-osmotic transport

The mechanism of this transport process is not very clear. Delivery of the medication via the oil membrane takes place in the presence of the light.

a) Solubilization of internal phase in the oil membrane

This is a noticeable transmission system. Within a stabilization of minute quantities of a internal phase throughout the membrane phase results within transfer of very comparatively tiny quantities of materials.

4. Stability of multiple emulsion: ¹³

It is a phenomenon which depends upon equilibrium between three phases; water, oil and surfactant. Nevertheless, multiple emulsions are thermodynamically unstable. A little emulsifier may result in unstable systems, whereas too much emulsifier may lead to toxic effects and can cause destabilization. A few other mechanisms have been described that also contributes to instability of multiple emulsions:

I. Coalescence of multiple emulsion droplets or internal droplets.

II. Puncture of oil layer on exterior of inner drops.

III. Shrinkage as well as swelling of inner droplets due to osmotic gradient across the oil membrane.

IV. Flocculation of internal aqueous phase and multiple emulsion droplets and Phase separation.

The main problem in regards to stability is the presence of two interfaces which are thermodynamically unstable. So, two different emulsifiers are necessary for their stabilization; one with a low HLB (Hydrophile-Lipophile Balance) value for W/O interface and the another one with a high HLB value for O/W interface. We can stabilize the emulsions by using electrolytes, by forming polymeric film, by interfacial complexation between non-ionic surfactant and macro molecules. Following techniques we might use to resolve destabilization in multiple emulsions:

4.1. The inner phase

We can stabilize the inner W/O emulsion mechanically, or in presence of better emulsifiers, reducing its droplet size. Also we can achieve our aim by preparing microspheres and increasing the viscosity of inner water.

4.2. The oil phase

Modifying the nature of the oil phase by increasing their viscosity, by adding carriers or by adding complexing agents to the oil.

4.3. The interfaces

This can be done by stabilizing inner and/or outer emulsion by using polymeric emulsifiers, macro molecular amphiphiles or colloidal solid particles to form strong as well as more rigid film at the interface; also by in-situ polymerization at the interface.

Hence, stability of multiple emulsions can be improved by forming a polymeric film or macro molecular complex across the oil/ water interfaces.

5. EVALUATION OF MULTIPLE EMULSIONS ^{2, 3, 4}

5.1. Entrapment Efficiency

The percentage entrapment efficiency is essential for the determination percentage content of active substance. The Percentage entrapment efficiency (% pee) was determined by taking freshly prepared W/O/W multiple Emulsions and immediately centrifuged at 4000rpm for 10min. Then 1 ml of a Aqueous phase (a lower layer) has been precisely removed through 2 ml hypodermic syringe as well as distilled adequately with phosphate buffer 6.8. The solution was filtered through a Millipore filter (0.22mm in pore size) and drug content was analyzed on UV spectrophotometer at 275nm. The Encapsulation Efficiency was determined by the following equation:

% EE = (Total drug incorporated – Free Drug) / Total drug * 100

5.2. Globule Size or Particle Size

In this study, globule sizes of the multiple emulsions were determined by using Zeta Analyzer Apparatus, Zeta Analyzer is an apparatus mainly used for the determination of particle size, Zeta Potential, molecular weight are primarily based on Light transmittance and Scattering Phenomenon.

5.3. In Vitro Release Studies

The in vitro medicine release analysis was conducted out on such a simple disintegration cell utilising cellophane cell wall (thickness-200 mm, breaking strength-2.7 kgf / cm). Prior to release experiments, the cellophane membrane has been soaked in water besides 6 hours, rinsed regularly 4 times by modifying distilled water, then immersed in 5% v / v glycerol solution for at least 60min and rinsed finally to 5 portions of distilled water. 15 ml freshly formulated multiple emulsion has been added to the donor chamber, mainly made of such a hollow glass tube (2.5 cm in diameter and 10 cm in length) as well as the membrane has been attached on the bottom end of the tube with a nylon string. This tube was dipped into a 250ml vessel containing 100ml of PBS pH 6.8 and was stirred at 100 rpm on a magnetic stirrer and maintained at 37°C which acted as receiving chamber. Aliquots of 1 ml had been collected from having received chamber at fixed time intervals and the drug loading had also been determined on UV spectrophotometer at 275 nm after suitable dilution.

5.4. Rheological analysis¹⁵

The rheological behavior of the emulsions can evaluate utilizing cone as well as plate geometries (60 mm diameter; 2 ° cone angle; 0.105 mm gap) on even a Rheo Stress 75 (Haake, Germany) rheometer with a regulated temperature managed by either a flowing water bath (DC5, Haake) and a Peltier but also kept at 25 ° C. Measurement can be done at 24 h, 15 days and 20 days after the sample preparation. Data collection, treatment and regression can perform on RheoWin 3 (Haake) softwareSince adding into rheometer, the emulsion remained unperturbed for 10 min even before following tests to always be perform:

I. Flow curve (γ˙=0.1−100 s−1; t = 200 s), where the coefficient of determination R² was used as a parameter for the choice of the rheological model adopted.

II. Stress ramp (σ = 0.01–70 Pa; 10³ s), where the yield stress value can establish at a breakpoint in the slope of two power law regressions in a deformation, γ, versus stress, σ, double logarithmic plot. The intercept of the two curves are calculate by software (Kutschmann, 2003).

III. Dynamic stress sweep (σ = 0.01–100 Pa; f = 1 Hz).

IV. Dynamic frequency sweep (f = 0.01–10 Hz; σ = 1.0 Pa).

V. Creep–recovery test (σ = 2.5 Pa; 300 s; σ = 0 Pa; 300 s) and (σ = 5.0 Pa; 300 s; σ = 0 Pa; 300 s) for emulsions and (σ = 0.05 Pa; 300 s; σ = 0 Pa; 300 s) for polysaccharide solutions (3.5%, w/w) in which the compliance, plot J, versus time and the recovery, γ_E/γ_MAX, of deformation, γ, determine (γ_E is the elastic deformation and γ_MAX is the maximum deformation).

5.5. Drug Release Studies¹⁶

Two-compartment diffusion cells were used. The top (donor) and the bottom (receptor) compartments had volumes of 8 and -600 ml, respectively. The medical grade poly(dimethylsi1oxane) membrane, 0.127-mm thick, was placed between the formulation and the sink compartment giving an interfacial area of -7.10 cm2. Except when otherwise stated, 0.1 M HCI was used as the receptor phase to ensure sink conditions for the local anesthetic bases within the receptor compartment. The cell was submerged in a 32°C water bath. In addition, experiments with aqueous solutions were carried out at 25 and 37°C. The sink mixture has been pumped via a spectrophotometer (Zeiss DM4 double-beam spectrophotometer) by the a peristaltic pump (Gilson Minipuls 11, Isoversing tubing). All tubing was made of teflon and the sink was agitated with a magnetic stirrer at a calibrated speed of 600 rpm. When the temperature of the sink had been regulated, 8 mL or grams of the formulation were placed in the donor compartment, and the automatic recording of the absorbance of the sink at 230 nm was started. Whenever the proportion of lidocaine and prilocaine throughout the sink was not recognised, an HPLC technique was used to measure lidocaine and prilocaine independently. The L-P flux was calculated by summing the fluxes of the individual species. The release studies were carried out with and without stirring of the formulations. The stirring was carried out with a glass blade at a precalibrated speed of 140 rpm, the speed at which the release rate reached its maximum when tested up to 200 rpm.

6. APPLICATION OF MULTIPLE EMULSIONS ^{5, 6, 11, 19}

6.1. Multiple emulsions in cancer therapy

Most chemotherapeutic agents are being used as emulsions because they are water-soluble. In the manner of an emulsion it will be easy to adjust release rates of drug and inhibit strong side effects of the drug. A single emulsion could not be used even though W / O emulsions usually have a high viscosity which infusion of emulsions to arteries / blood vessels via catheters would be challenging. Also O/W emulsions aren't just an option because they don't really encapsulate the drug. But W/O/W emulsion systems are suitable drug carriers because of the encapsulation of the drug in the internal water phase and the low viscosity due to the external water phase.

6.2. Multiple emulsions in herbal drugs

Apart from its targeted sustained release, forming the herbal drug into emulsion will also be strengthen the stability of the hydrolyzed materials, improve the penetrability of drugs to the skin and mucous, as well as reduce the medicine stimulus to the tissues. Thus far and, a few forms of herbal drugs, like camptothecin, Bruceajavanica oil, coixenolide oil and zedoary oil have been made into emulsion.

6.3. Vaccine/vaccine adjuvant

The Use of such w / o / w multiple emulsion also as modern concept of adjuvant for antigen had first been reported by Herbert. Such emulsions elicited greater autoimmune reaction than antigen alone. Rishendra and Jaiswal established a multiple emulsion vaccine against Pasteurella multocida infectious disease in cattle. This immunization made a contribution all these humoral as well as cell-mediated immune responses in safety against the infection.

6.4. Oxygen substitute

A multiple emulsion of aqueous oxygen carrying material in oil in outer aqueous phase is suitable for provision of oxygen for oxygen transfer processes. Hemoglobin multiple emulsions throughout physiologically appropriate oil inside an external aqueous saline solution has been offered in relatively low droplet size and provide oxygenation through blood vessels to required body tissues as well as organs while also supplying a blood substitute.

6.5. Inverse targeting

Regarding this approach Talengaokar and Vyas were prepared poloxamer 403 containing sphere in-oil-water (s/o/w) multiple emulsion of Diclofenac sodium by gelation of inner aqueous phase and they examined the effect of poloxamer 403 on surface modification for inverse targeting to reticulo endothelial system-rich organs. The results concluded that even this multiple system containing poloxamer will have capability to retards the RES uptake of drugs mainly to liver, brain and trying to target to non-RES tissue like lungs, inflammatory tissue.

6.6. Multiple emulsions in diabetes

Toorisakaet.al. Established a S/O/W emulsion besides oral delivery of insulin. Surfactant-coated insulin has been dispersed within oil by ultrasonication, one such dispersion has been blended with the exterior water phase with such a homogenizer and after that, the S/O/W emulsion thus obtained is now re balanced to such a continuous particle size by passage through SPG membrane. The S/O/W emulsion showed hypoglycemic activity for a long period after oral administration to rats.

6.7. Multiple emulsions in food

The potential application of the double emulsions is now the food industry. Preliminary studies have been performed in the field of entrapment of a flavor component in a release system. Vulnerable food items and flavorings could be encapsulated in W/O/W emulsions. Perceptual evaluations have demonstrated there is a significant taste difference among W/O/O/W emulsion and O/W emulsions containing the same ingredient, and there is a put on hold to discharge of flavor in double emulsion.

6.8. Drug over dosage treatment

This scheme could even be developed for the over medication treatment whilst also using the variation in the pH For example- barbiturates. In such emulsions, the inner aqueous phase of emulsion does have the basic buffer and then when emulsion is taken orally, acidic nature of the stomach behaves as an external aqueous phase. In the acidic component barbiturate remains mainly in unionized form that also transmit via oil membrane in to the inner aqueous medium and will get ionized. Ionized medication has much less affiliation to pass the oil membrane while also getting entrapped. There by, entrapping excess medication in multiple emulsions cure over dosage.

6.9. Taste masking

Multiple emulsions of chloroquine, an anti malarial agent has also been prepared successfully and was discovered to mask the sour after taste efficiently. Taste masking of chlorpromazine, an antipsychotic medication also has been reported by multiple emulsions.

7. CONCLUSION:

The Multiple Emulsion is one of the innovative methods of drug delivery for the improvement of the different drug properties, such as bioavailability, flavour, release time, etc. The inventions include new innovative formulations for improving the administration of the medication and improving the palatability of the drug by adding them into the different formulations. The Multiple Emulsion is a dynamic polydispersed system of multiple emulsions. An emulsion that is contained in another emulsion that can be found in several applications such as taste masking, prolonged release, distribution of the unstable drug and environmental prevention of the drug, etc.

8. REFERENCES:

1. Navneet Kumar Verma1, Jai Narayan Mishra, Multiple emulsions and its stabilization: A review, International Journal of Chemistry Studies, 2019; (3)3: 12-16.

2. Raynal S, Grossiord JL, Seiller M, Clausse D, A topical W/O/W multiple emulsion containing several active substances: formulation, characterization and study of release, Journal of Controlled Release, 1993; 26(2): 129-140.

3. Jong-wook Ha And Seung-man Yang., Breakup of A Multiple Emulsion Drop In A Uniform Electric Field., Journal of Colloid and Interface Science, 1999; 213: 92–100.

4. Sagar Savale1, Uday Mali, Patil Prafulla, Formulation and Evaluation of W/O/W MultipleEmulsions with Diclofenac Sodium, International Journal of Pharmacy and Pharmaceutical Research, 2016; 6(2): 122-134.

5. Rajeshkumar, Murugesan Senthilkumar, Nanjaian Mahadevan, Multiple emulsions: A Review, International Journal of Recent Advances in Pharmaceutical Research, 2012; 2(1): 9-19.

6. Sumana Khosh, Formulation and characterisation of multiple emulsions with various additives, International Journal of Research in Pharmaceutical and Biomedical Sciences, 2011; 2(2): 751-759.

7. Sivapriya S., Daisy P. A, Boby Johns G., Praveen Raj R., Noby Thomas and Betty Carla, multiple emulsions a coprehensive review, World Journal of Pharmaceutical and Medical research, 2016; 2(5): 83-88.

8. Lachman, L, Lieberman, H.A., The Theory and Practice of Industrial Pharmacy, CBS Publishers and Distributors, 2009, 502-32.

9. Neeraj Bhatia, Siddharth Pandit, Shikha Agrawal, Dishant Gupta, A Review on Multiple Emulsions, International Journal of Pharmaceutical Erudition, 2013; 3(2): 22-30.

10. Sarika S. Lokhande, Namita N. Phalke, Vijay N. Raje, Savita S. More, An Update Review on Recent Advancements in Multiple Emulsions, International Journal of Research and Scientific Innovation, 2018; 5(8): 90-96.

11. Bhatia N., Pandit S., Agrawal S. and Gupta D, A review on multiple emulsions. International Journal of Pharmaceutical Erudition, 2013; 3(2): 22-30.

12. R. M. Mehta, pharmaceutics II Book, Vallabh Prakashan, 4th Edition, 2015, 153-161.

13. Navneet Kumar Verma, Jai Narayan Mishra, Multiple emulsions and its stabilization: A review, International Journal of Chemistry Studies, 2019; 3(3): 12-16.

14. Asuman Bozkir and Gokhan Hayta, Preparation and Evaluation of Multiple Emulsions Water-in-oil-in-water (w/o/w) as Delivery System for Influenza Virus Antigens, Journal of Drug Targeting, 2004: 12(3): 157–164.

15. Ricardo Padilha Vianna, Camera lucia Oliveira Petkowicz, Joana Lea Meira Silveira, Rheological characterization of O/W emulsions incorporated with neutral and charged polysaccharides, carbohydrate polymaer on Science Direct, 2013; 93(1): 266-272.

16. Adela A. Nyqvist-Mayer, Arne F. Brodin, Sylvan G. Frank, Drug release studies on an oil-water emulsion based on a eutectic mixture of lidocaine and prilocaine as the dispersed phase, Journal of Pharmaceutical Sciences, 1986; 75(4):365-373.

17. Azhar Yaqoob Khan, Sushama Talegaonkar, Zeenat Iqbal, FarhanJalees Ahmed and Roop Krishan Khar, Multiple Emulsions: An Overview, Bentham Science Publishers Ltd, 2006; 3(4): 429-443.

18. Anisha Agrawal, Sunisha Kulkarni, Shyam Bihari Sharma, Recent advancements and applications of multiple emulsions, International Journal of Advances in Pharmaceutics, 2015; 4(6): 94-103.

19. Thorat S Sheela, Dadde S Gurunath, A Mohite Swapnali, R Chavan Rohan Kumar, D Mali Ramling., Comparative Study of Antibacterial Activity of Tamarindus indica and Tagetes erecta, Research Journal of Pharmacognosy and Phytochemistry, A and V Publications, 2019; 11(3): 186-188.

20. Raut I.D. Dhadde G.S, Yadav J.P, Sapate R.B., Mali H.S, In vitro anthelmintic activity of crude extract of flowers of Bougainvillea Spectabilis Wild against Pheretima Posthuma, International Journal of Pharmacy and Pharmaceutical Research, 2020; 17: 1-18.

21. Snehal Chandanshive, Akanksha Jagdale, Sandeep Patil, Hanmant Mali, In-Vitro Antispasmodic Efficacy of Ethanolic Extract of Leaves of Sesbania Grandiflora, World Journal of Pharmaceutical Research, 2020; 9(2): 915-921.

22. Dhadde Gurunath S., Mali Hanmant S., Sapate Rohit B., Vakhariya Rohan R., Raut Indrayani D., Nitalikar Manoj M., Investigation of In-Vitro Anthelmintic Activity of Methanolic Extract of Tylophora Indica Leaves Against Haemonchus Contortus, Journal of University of Shanghai for Science and Technology, 2021; 23(2): 37-42.

Received on 27.02.2021 Modified on 20.03.2021

Asian Journal of Pharmacy and Technology. 2021; 11(2):156-162.

DOI: 10.52711/2231-5713.2021.00026