1. INTRODUCTION:

Immunodeficiency is a state in which the human body system is unable fight to external environmental factors such as temperature, climate condition, some drugs, such as steroids, can be either an adverse effect or the intended purpose of the treatment, inherent deficiency and microbial organism, which affect the patient's immune system. In Indian sub continent Guava (Psidium Guajava) leaf and Pomegranate (Punica granatum) are fairly wild growing, middle size evergreen tree naturally reaching 10–25 ft in height, consisting of sleek leaves (Torane Rasika Charudatta et. al 2015) cultivated throughout the hotter parts of India (Praveen D, 2016).

The neem (Azadirachta indica) leaves contain ingredients such as nimbin, nimbanene, 6-desacetylnimbinene, nimbandiol, ascorbic acid, n-hexacosanol and amino acid, 7-desacetyl-7-benzoylazadiradione, 7-desacetyl-7-benzoylgedunin, 17-hydroxyazadiradione, and nimbiol (Hossain M. A, 2011, Robinka Khajuria, 2014, Stefanello M.E.A, 2008) has

been used as traditional folk medicine and also provides many environmental services. In herbal homes, leaves of this plant are used for the treatment of hepatic disorders, hypertension, and diabetes (Oluwatosin Adekunle Adaramoye et.al 2014).

A. heterophyllus exhibit numerous medicinal properties such as antibacterial, antioxidant, anti-diabetic, anti-inflammatory, anti-diuretic, and have been useful in the treatment of fever, skin diseases, convulsions, constipation, ophthalmic disorders and snake bite (Shirajummunira et.al 2014, Sureka, R et.al 2019). By this we can understood that the cancer also comes from this immunodeficiency so we overcome this by the using of natural medicine such as herbal medicine because there is solution for all problems in the natural medicine in this to prepare the anti-immunodeficiency tablet we take the neem leaf, guava leaf and the high vitamin content pomegranate leaf. Then extract the phytochemicals and mix all the three leaf phytochemicals and used as an anti-immunodeficiency tablet.

2. MATERIALS AND METHODS:

The plant leaves were collected from the Local area of Erode District, India during September. The Collected plant leaves were cleaned with normal water and then rinsed with double distilled water (M. M. Tajkarimi, 2010 and G. G. F. Nascimento,2000). After washing, the material is air-dried at room temperature in a well ventilated room. The dried leaves are grinded to fine powder form.

2.1 Phytochemical extraction:

The ingredient present in the all three leaves has been extracted through the process of infusion, which a method of extraction is concerning boiling the plant material—and percolation, in which water is passed through the material (S. Chanda, 2011 and Ahmad I, 1998). Infusion is a very easy chemical process used with botanicals to dissolve or release their active ingredients in water, oil, or alcohol. In our experiment water has been used as an extracting agent.

2.2 Guava (Psidium Guajava) leaf extraction:

The sample guava leaf is taken 10grams leaves by weighing in the digital weight balance then it is mixed with the one litre distilled water and then it is started infused for 20 minutes for the temperature at 120^oC after it is infused allowed to stand for five minutes then it is filtered by the Whatman filter paper and the filtrate and the solution is separated and the solution is taken for further proceedings as per standard procedure.

2.3 Pomegranate (Punica granatum) and neem (Azadirachta indica) leaf extraction:

The sample neem leaf is taken as 6grams and the pomegranate leaf 4 grams is taken as 10grams both of them are weighed in the weighing balance for the correct proportion and both of them are mixed with the one litre distilled water and then it is infused for 25 minutes at the temperature of 125^oC after it is infused then it is filtered (Biswas K et.al, 2002 and Tarasankar Maity et al, 2012) by the Whatman filter paper and separate the solution and filtrate. The solution is taken for the further proceedings.

2.4 Composition-1 herbal drug compound preparation:

The prepared guava leaf extract solution is taken 150ml of the solution and the mixed neem and pomegranate leaf extract solution is taken as 100ml of the solution. For the proper mixing of the two samples to produce the herbal drug compound we kept in the magnetic stirrer for the 45 hours at the 500rpm.

2.5 Composition-2 herbal drug compound preparation:

The prepared guava leaf extract solution is taken for the amount of 200ml of the solution then from the mixed sample extract such as neem (Jacobson M,1990) and the pomegranate leaf extract solution taken for amount of 150ml of the solution and also they are additionally mixed with the zinc nitrate solution for the 10ml (solution) then it is kept in the magnetic stirrer for the 45 hours at the 550rpm for the production of the new herbal drug compound for the pharmaceutical applications.

2.6 Precipitate formation and separation:

The composition 1 solution is taken after 45 hours stirring and then it is allowed to settle for one day after the precipitate is settled down by the sedimentation technique it settled precipitate is separated from the solution by using the Whatman filter paper and then take the precipitate 1. The composition 2 solutions is taken after stirring it is also allowed to settle the precipitate after they are settled they are filtered by the Whatman filter paper and take the precipitate 2. After that the precipitate has been kept in dryer for 3 hrs at 100^oC to remove the moisture present in the samples.

3 RESULTS AND DISCUSSION:

3.1 XRD analysis of the sample:

In order to find out the main components and their existing phase in the powder, X-ray diffraction (XRD) analysis was carried out on a computer controlled diffractometer (Surendran and Baral, 2018 and Shyam R et. al, 2017). XRD analysis pattern showed the peaks related with the main component for the both samples.

Table 1: Composition of the Samples

Composition of the Sample No:1
Score	Compound name		Chemical formula	Crystal structure
21	[1, 7, 9 - Tris(dimethylsulfide) - Dodecahydrononacloso-borane] Tetrafluoroborate		C₆H₂₇B₁₃F₄S₃	Anorthic
31	Fluoro-deca(closo)borane		H₁₃B₁₀F₁	Monoclinic
33	Bismuth Niobium Oxide (2.94/1/7)		Bi_2.938Nb₁O₇	Tetragonal
29	Aza-closo-decaborane		H₁₀B₉N₁	Monoclinic
30	Decaborane-14		H₁₄B₁₀	Monoclinic
23	Tetramethylammonium Tetraaquadifluorotitanium (III) Hexafluorotitanate Hydrate		C₄H₂₂F₈N₁O₅Ti₂	Orthorhombic
26	Magnesium Bis(tetradeuteridoborate)		D₈B₂Mg₁	Orthorhombic
Composition of Sample No: 2
62		Lithium Manganese Oxide (1/2/4)	Li₂Mn₂O₄	Orthorhombic
49		Mue-(N,N-Dimethylamido) pentahydro(trimethylphosphide)-diboron	C₅H₂₂B₂N₁P₁	Orthorhombic
39		Tricesium Heptaborate	B₇Cs₃O₁₂	Monoclinic

Table 2: FTIR Analysis of Samples:

IR Peaks	Bond	Possible Functional groups	IR Peaks	Bond	Possible Functional groups
Sample No 1			Sample No 2
3310.74	–C≡C–H: C–H stretch	alkynes (terminal)	3299.53	–C≡C–H: C–H stretch	alkynes (terminal)
2921.36	C–H stretch	alkanes	2928.11	C–H stretch	alkanes
1665.05	C=O stretch	carbonyls (general)	1802.39	C=O stretch	carbonyls (general)
1331.89	C–N stretch	aromatic amines	1334.81	N–O symmetric stretch	nitro compounds
1147.02	C–N stretch	aliphatic amines	1147.43	C–N stretch	aliphatic amines
858.13	N–H wag, C–H “oop”	1°, 2° amines, aromatics	1076.80	C–N stretch	aliphatic amines
760.96	C–Cl stretch	alkyl halides	926.33	O–H bend	carboxylic acids
			856.98	N–H wag, C–H “oop”	1°, 2° amines, aromatics
			761.74	C–Cl stretch	alkyl halides
			644.66	C–Br stretch	alkyl halides

Fig 3: FTIR Analysis of Sample 1

Fig 4: FTIR Analysis of Sample 2

3.3. FESEM with EDAX of the samples:

In order to find out the morphological structure of the samples, the powdered material was analysed by the Scanning electron microprobe images are at 500X magnification. From the micrograph images, it was observed that there is a change in surface morphology for the both samples (Goswami, L et al, 2017). For the sample 1 the structure was in irregular porous structure and for sample 2 it shows that not much of porous. From the EDAX analysis, it is observed that there is a different mass percentage for both the samples.

Fig 5: The FESEM image of sample 1 and 2

Fig 6: The FESEM image of composition of sample 1 and 2

Table 3: Composition of the Samples from EDAX

Sample No: 1			Sample No: 2
Element	Series	Wt %	Element	Series	Wt %
Carbon	K-series	49.14	Carbon	K-series	50.90
Oxygen	K-series	45.11	Oxygen	K-series	48.76
Zinc	K-series	5.71	Zinc	K-series	0.34
Phosphorus	K-series	0.04	Zinc	K-series	0.34

4. CONCLUSION:

The different types of phytochemicals are extracted from the various types of plants such as neem leaves, guava leaves and pomegranate leaves. Then they are taken as two different samples with different compositions (ratios) and they are stirred and filtered, then filtered precipitate is now mixed with the starch in the two samples at same level to increases the medicinal activities. Then the different types of samples are dried at normal condition then they are further analyzed by the x-ray diffraction, Fourier Transform Infrared (FTIR), FESEM with EDAX, these characterization are done and found the high content of zinc, carbon, oxygen and phosphorus and their functional group and structure such as orthorhombic, monoclinic, tetragonal. After in the antimicrobial activity analysis the micro-organisms such as staphylococcus aureus, Escherichia coli, salmonella typhimurium, candida spp., are used and analyzed from this the plant samples are rapidly reacted and kills most micro-organisms and shows the better antimicrobial activity. Then from these we have concluded that the plant samples are made as medicine for immunodeficiency diseases and reduce the risk of this deficiency and increases the healthy life of most peoples and the economic value of health of our world. This may help to serve as selective agent against some of immunodeficiency diseases.

5. ACKNOWLEDGEMENT:

We are thankful to the PSG College, Coimbatore, India for their help to carry out sample analysis XRD, FTIR, FESEM. The authors also wish to express their grateful to professors and friends who made this study possible.

6. CONFLICT OF INTEREST:

The authors declare no conflict of interest.

7. REFERENCES:

1. Torane Rasika Charudatta, Ruikar Anjali Deepak, Deshpande Nirmala Ramchandra, Evaluation of Phenol Content from a Medicinally Important Plant – Artocarpus Heterophyllus, Int. J. Pharm. Sci.Rev. Res., 2015, 33(1), 37-39. http://globalresearchonline.net/journalcontents/v33-1/09.pdf.

2. Praveen D, Ranadheer Chowdary P, Gandikota Thanmayi, Gangabathina Poojitha, Vijey Aanandhi M Antioxidant and Analgesic Activity of Leaf Extracts of Artocarpus heterophyllus, Research Journal of Pharmacy and Technology, 2016, vol: 9, page:3. doi: 10.5958/0974-360X.2016.00047.0

3. Hossain M. A., Shah M. D., Sakari M. Gas chromatography–mass spectrometry analysis of various organic extracts of Merremia borneensis from Sabah. Asian Pacific Journal of Tropical Medicine. 2011, vol: 4(8), page: 637–641. doi: 10.1016/s1995-7645(11)60162-4.

4. Robinka Khajuria, Loveleen Kaur, Aditi Kaushik, Gurpreet Saredia. Evaluation of Antimicrobial and Phytochemical Properties of some Indigenous Indian plants. Res. J. Pharmacognosy and Phytochem. 2014, vol.6(1),page:5,doi:indianjournals.com/ijor.aspx?target=ijor:rjpp&volume=6&issue=1&article=002

5. Stefanello M.E.A, Cervi A. C, Ito I. Y, Salvador M. J, WisniewskiJr A, and Simionatto E. L., Chemical composition and antimicrobial activity of essential oils of Eugenia chlorophylla (Myrtaceae), Journal of Essential Oil Research,2008, vol.20,no.1,page:75–78. doi.org/10.1080/10412905.2008.9699427.

6. Oluwatosin Adekunle Adaramoye and Olubukola Oyebimpe Akanni, Effects of Methanol Extract of Breadfruit (Artocarpus altilis) on Atherogenic Indices and Redox Status of Cellular System of Hypercholesterolemic Male Rats, Adv Pharmacol Sci., 2014: 605425. doi: 10.1155/2014/605425.

7. Shirajummunira, Antioxidant and anti diarrhoeal activities of methanolic extract of Artocarpus heterophyllus seed, Int. J. Pharm Drug Anal., 2014, Vol: 2, Issue:11, Pages:890-895. doi: 0.5958/0974-360X.2016.00047.0

8. Sureka, R. Dhanalakshmi, C. Mary Sharmila, R. Chithra Devi. Comparative Analysis on Significance of Purification Process of Monosilai in Siddha medicine using Modern Instrumental Techniques: Standardization Approach. Asian J. Pharm. Ana. 2019; vol: 9(2), page: 99-104. https://doi.org/10.5958/2231-5675.2019.00019.X

9. Tajkarimi M. M., Ibrahim S.A., and Cliver, D.O., Antimicrobial herb and spice compounds in food, Food Control, 2010, vol. 21, no. 9, page: 1199–1218. https://doi.org/10.1016/j.foodcont.2010.02.003.

10. Nascimento G. G. F, Locatelli J, Freitas P. C, and Silva G. L., Antibacterial activity of plant extracts and phytochemicals on antibiotic-resistant bacteria, Brazilian Journal of Microbiology, 2000, vol. 31, no. 4, page. 247–256. http://dx.doi.org/10.1590/S1517-83822000000400003.

11. Chanda S. and Kaneria M., Indian nutraceutical plant leaves as a potential source of natural antimicrobial agents in Science against Microbial Pathogens: Communicating Current Research and Technological Advances, A. Mendez-Vilas, Ed., Formatex Research Center, 2011, vol. 2, pp. 1251–1259. doi: 10.1155/2013/746165.

12. Ahmad I, Mehamood Z and Mohammad F Screening of some Indian medicinal plants for their antimicrobial properties. J. Ethnopharmacol. 1998, vol 62, page: 183–193. doi: 10.1016/s0378-8741(98)00055-5.

13. Biswas K, Chattopadhyay I, Banerjee R. K and Bandyopadhyay U, Biological activities and medicinal properties of neem (Azadirachta indica), Curr. Sci., 2002, vol: 82, page:1336–1345. https://www.jstor.org/stable/24106000.

14. Tarasankar Maity, Ayaz Ahmad, Nilanjan Pahari, Subarna Ganguli. A Review on Hepatoprotective herbs for treatment of various liver Disorders. Research J. Pharm. and Tech. 5(5): 2012; Page 602-607. https://rjptonline.org/AbstractView.aspx?PID=2012-5-3-25

15. Jacobson M, Review of neem research in the United States. In: Locke, J.C. and Lawson, R.H. (eds) proceedings of a workshop in neems potential in pest management program. USDA-ARS. Beltsville, MD.ARS-86. 1990, page: 4-14.

16. Surendran G, Baral S.S, Biosorption of Cr(VI) from wastewater using Sorghastrum Nutans L. Nash, Chem.Ecol., 34(8) (2018) 762-785. https://doi.org/10.1080/02757540.2018.1487407.

17. Shyam R. Annapure, Rahul A. Waghmare, Shantilal D. Rathod, Narayan P. Adlinge. Antioxidant, Antimicrobial Study of Synthesized and Characterized metal Complexes of Mn (II), Fe (III), Co (II) and XRD, Thermal Study. Asian J. Research Chem. 2017; 10(4):541-545. https://doi.org/10.5958/0974-4150.2017.00089.X

18. Baral S.S., Surendran G, Das, N, Rao P.V, Statistical design of experiments for the Cr(VI) adsorption on weed salvinia cucullata, Environmental Engineering and Management Journal., 2013;12: 465-474. doi.10.30638/eemj.2013.058.

19. Samer Housheh, Saleh Trefi, M. Fawaz Chehna. Identification and Characterization of Prasugrel Degradation Products by GC/MS, FTIR and 1H NMR. Asian J. Pharm. Ana. 2017;7(2): 55-66. https://doi.org/10.5958/2231-5675.2017.00010.2

20. Goswami L, Kumar R.V, Arul Manikandan N, Pakshirajan K and Pugazhenthi G. Anthracene biodegradation by oleaginous Rhodo coccus opacus for biodiesel production and its characterization. Polycyclic Aromatic Compounds. 2017, 1–13. doi.org/10.1080/10406638.2017.1302971

Received on 22.07.2020 Modified on 27.09.2020

Research J. Pharm. and Tech. 2021; 14(6):3244-3248.

DOI: 10.52711/0974-360X.2021.00564