Formulation and evaluation of topical spray containing standardized senna alata leaf extract

pharmacy project

1. KLE COLLEGE OF PHARMACY,HUBBALLI FORMULATION AND EVALUATION OF TOPICAL SPRAY CONTAINING SENNA ALATA LEAF EXTRACT FOR ODOU R CAUSING BACTERIA Name of the students: Rakshath Naik (NA0220038) Naveed M.K. (NA0220032) IV B. Pharm (8 th Semester) Guided by Dr. Vijaykumar meti Assistant Professor, Dept of Pharmaceutics KLE College of Pharmacy, Hubballi

2. CONTENTS ABSTRACT INTRODUCTION NEED FOR STUDY AIM OBJECTIVES MATERIALS AND METHODS RESULTS CONCLUSION REFERENCES

3. ABSTRACT Axillary malodour is a frustrating condition for many people. It can lead to significant discomfort and various psychological effects. The underarm microbiome plays a major role in axillary malodour formation. Bacteria living in the epidermis, sweat glands, sweat pores and hair follicles play a pivotal role in malodour development. The herbal medicines have shown complementary efficacy against wide range of microbial infections that may even inhibit the growth of odour causing bacteria. Senna alata extracts are potent antibacterial. The objective of the study was to standardize the contents of Senna alata leaf extract and develop formulations of Senna alata topical spray to inhibit the growth of odour causing bacteria. The process includes extraction , qualitative analysis by TLC, antimicrobial activity, pH, washability , dryability , spray angle and spray pattern. The formulation showed darkish brown in colour , good washability , with pH 6 - 6.3 and were homogeneous in nature and the drying time of the non-film forming spray is within 7.2-8mins. All the parameters which are been evaluated are under the desired range. Thin Layer Chromatographic analysis confirmed active ingredients like rhein , and aloe- emodin from leaf extract. Pure extract showed higher antibacterial activity and increase in concentrations of extract shown increase in antimicrobial activity. Spray pattern and Spray angle were uniform in nature and were in desired range. Senna alata topical spray preparation shown increased antimicrobial efficiency compared to pure Senna alata leaf extract.

4. Introduction Axillary malodour is a frustrating condition for many people. It can lead to significant discomfort and various psychological effects . The underarm microbiome plays a major role in axillary malodour formation. Bacteria living in the epidermis, sweat glands, sweat pores and hair follicles play a pivotal role in malodour development .

5. There are two different sweat glands responsible for secreting perspiration. Research on natural plants has drawn a lot of interest worldwide with new developments in herbal research to determine the ability of natural medicines used in traditional and complementary medicines for human diseases. Types of glands Eccrine glands Apocrine glands Function Control body temperature Produce m ore salty sweat Axillary malodour which yields foul smell Types of sweat glands and their function : Continued….

6. Eccrine glands and apocrine glands are the two sweat glands meant for axillary malodour . Eccrine glands secret salts and water that reach the skin through coiled ducts or tubes when the nervous system senses the body heat. Apocrine glands contains organic substances (fatty substances). These glands secret through the hair follicles and present large number in armpits (2-4). Fig 1: Sweat glands

7. Bacteria’s Responsible for Axillary malodour Two different types of microbes found in the axillary region such as Staphylococcus spp and Corynebacterium spp . Bacteria’s are being more prevalent in males and contributing to a more pronounced body odour compared to female counterpart. Women have 75% more apocrine glands in their armpits than men, but male apocrine glands are larger and may be more active in order to supply nutrients for bacterial growth . Corynebacterium spp is responsible for the formation of a strong axillary malodour and Staphylococcus spp , Propionibacterium spp relatively contributes low level axillary odour formation .

8. PLANT DESCRIPTION: Fig 2: Senna alata Plant and Taxonomy Senna Alata is a shrub belonging to the Fabaceae family mainly found in tropical and subtropical area. Its commonly well known as a candle bush or ringworm tree and locally acquainted as Daun Gelenggang . Traditionally been used for the treatment of constipation and dermatophyte infections. Morphological description: height 3-4m, yellowish-green leaf, 50-80cm long leaves with 5-14 leaflet pair. Usually its grown in tropical region.

9. Need for Study Senna alata leaf extract are traditionally used to treat various skin associated diseases. Many reasearch work were done on the senna alata plant extract but most of them were based on analytical tehniques and pharmacological investigations. Senna alata soap bar, powder, cream, ointment formulations were approached by the researchers, but no research was done on topical spray as a safe antimicrobial preparation. Hence, standardization of senna alata leaf extract with respect to content, formulation and effcacy of topical spray are need to be approached.

10. AIM Formulation and evaluation of topical spray containing Senna alata leaf extract for odour causing bacteria . To standardize the content of Senna alata leaf extract. To formulate and develop Senna alata topical spray for odour causing bacteria . OBJECTIVES

11. Materials and Methods SL.NO Materials Manufacturer 1 Senna alata leaf Bio crops, Visakhapatnam 2 Ethanol Merck, Germany 3 Propylene Boxa chem,Goa 4 Aluminium chloride Moly chem, Mumbai 5 Glycerol Moly chem, Mumbai 6 Hydroxy propyl methy lcellulose Moly chem, Mumbai 7 Muller hintonagar/broth HI media, Maharashtra 8 DMSO Rankem, New Delhi 9 Fragrance oil Parag fragrance, Madhya pradesh

12. SL.NO Instument Manufacturer 1 Soxhlet Apparatus Sox-1LTR 2 Micro Peppet Hangzhou soy medTech CO.,LTD 3 Autoclave Servewell instruments, Bangalore 4 Incubator Servewell instruments, Bangalore 5 Sonicater Chem labs, Bangalore 6 Water Bath Chem labs, Bangalore 7 Electronic Balance Sartorius, Germany 8 UV visible Spectrometer Shimadzu, Japan EQUIPMENTS

13. Methodology Collection of Senna alata leaf, grounded to powder Extraction of Senna alata leaf using various solvents and concentrations by soxhlet extraction process Antimicrobial activity of standardized Senna alata extract against known antimicrobial strains Analysis of extract contents using Thin layer chromatography (TLC) Formulation development and in vitro antimicrobial activity of odour causing bacteria containing standardized Senna alata extract In vitro antimicrobial activity of formulation containing standardized Senna alata extract Evaluation of formulation will be carried out for the formulation containing standardized Senna alata extract

14. Extraction process

15. Authentication letter The collected Senna alata from Biocarp , Vishakpattanam , was identified and confirmed by Prof. Dr. Subhas Emmi, HOD, Department of Botony , KLE Societys , Shri Kadasiddeshwar Arts College & H S Kotambri Science Institute, Hubballi

16. Qualitative analysis by TLC The qualitative analysis of standard markers and senna alata leaf extract was carried out using TLC. Mobile phase - ethyl acetate:metanol:water (4:4:3) Stationary phase - precoated coated silica gel 60 F254(4.3×2cm) Standard solution of rhein , aloe- emodin , emodin were prepared by dissolving 1mg/ml of DMSO. 80% ethanolic senna alata leaf extract sample was prepared by dissolving 10mg/ml of DMSO

17. Antimicrobial activity test Method Cup plate method Microbial strains Staphylococcus epidermidis Media Muller Hinton agar Incubation temperature Method Cup plate method Microbial strains Staphylococcus epidermidis Media Muller Hinton agar Incubation temperature Senna alata leaf extract Concentration(mg/ml ) 10,20,30,40 and 50 Negative control DMSO Positive control Ciprofloxacin(0.2mg/ml) Senna alata topical spray Concentration(mg/ml ) 1%,3% and 5% Negative control Placebo

18. Formulation table of Senna aLata topical spra y SL.NO Ingredient Quantity Functions 1 Senna a lata leaf extarct 5%, 10% Antimicrobial 2 Propylene glycol 15ml Polyester compound 3 Aluminium chloride 15ml used as lubricant 4 Glycerol 10ml Thickening agent 5 Aquades 100ml vehicle 6 Hydroxyl methyl cellulose 2g Thickening agent 7 Perfume Q.S Fregrance

19. Evaluation of topical spray Physical examination - The formulation was evaluated for its appearance / clarity and homogeneity. pH Drying time Water washability Spray angle Spray pattern

20. Results 80% etanolic senna alata leaf extract Initial weight(g) Final weight percentage yield(%) Batch no. 1 25 4.62 18.48 Batch no. 2 25 4.35 17.40 Batch no.3 25 4.83 19.32 Average percentage yield 18.4% Extraction of S enna alata leaf Extraction of S enna alata leaf was performed using Soxhlet extraction process and obtaine d the yield 18.4%

21. Qualitative analysis of senna alata leaf extract by TLC SL.NO Sample UV detector Sample RF value Standard RF value 1 S enna alata 254nm 0.5, 0.86 0.5 Rhein , 0.8 Aloe- emodin , 0.9 Emodin TLC for S enna alata leaf extract The content of Senna alata leaf extract was qualitatively analysed by TLC and confirmed the presence of Rhein and Aloe emodin

22. Antimicrobial activity of S enna alata leaf extract using bacterial strain concentration (mg/ml) inhibition (in mm) 10 13.6 20 16.0 30 17.3 40 18.6 50 20.0 +ve control (Ciprofloxacin 0.2mg/ml) 37 -ve control (DMSO) 00 Senna Alata leaf extract Staphylococcus epidermidis Antimicrobial activity was performed for various concentration of Senna alata leaf extract using bacterial strain Staphylococcus epidermidis. The results confirmed increase in inhibition with increase in concentration.

23. Formualtion code colour pH Homogeneity Washability Texture Drying time F1 Darkish brown 6 Homogeneous Good Smooth 7.2min F2 Darkish brown 6.3 Homogeneous Good Smooth 8mins Evaluation of colour , pH test, Homogeneity, washability , texture and Drying time of spray

24. Evaluation of spray pattern Formulation spray pattern Average Ovality test F1 3.1 3.2 3.1 3.13 1.04 F2 3.4 3.5 3.4 3.43 1.03 Spray pattern

25. Evaluation of spray angle Formulation Spray angle F1 26 F2 22 Spray angle

26. Conclusion Senna alata leaf extracts shows potential antibacterial activity. Axillary malodour is a frustrating condition for many people. It can lead to significant discomfort and various psychological effects. The underarm microbiome plays a major role in axillary malodour formation. Bacteria living in the epidermis, sweat glands,sweat pores and hair follicles play a pivotal role in malodour development. The herbal medicines have shown complementary efficacy against wide range of microbial infections that may even inhibit the growth of odour causing bacteria. Secondary metabolites like Rhein and aloe emodin were found in Senna alata leaf extract that may act as antimicrobial activty . A spray containing Senna alata leaf extract was successfully developed for the prevention of odour causing bacterias . Physical evaluations showed acceptable appearance, pH, washability , dryability , spray patterns, spray angles etc. Theresults indicated that Topical spray containing the Senna alata leaf extractshowed higher antimicrobial activity compared to pure Senna alata leaf extract. .

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