An Investigation to Analyze the Efficacy of different Extracts of Omakai Seeds in Combating Parasitic Worms, (i.e. Anthelmintic Study) through In vitro Experiments
Anuradha Khopade*, Yash Bhusari, Atharv Mahakode, Khushi Wagh, Chandrakant Pimpalkar
Department of Pharmaceutical Science, Dr. Rajendra Gode Institute of Pharmacy, Amravati.
*Corresponding Author E-mail: anuradhakhopade2817@gmail.com
ABSTRACT:
The simplest way to control a life-threatening infection in humans and other living stalks is to use potent Anthelmintic medication, but there are few Anthelmintic medications on the market, and those that are available cause resistance during treatment in most helminth species. As a result, we will test the Anthelmintic properties of Omakai seeds for controlling life-threatening infections in humans and other living stalks. The Omakai seeds were saved for extraction, and the heat maceration process was used. The resulting extract was submitted to phytochemical screening, with tests for alkaloids, saponins, tannins, anthraquinone glycosides, reducing sugar for glycosides, and flavonoids performed according to standard technique. For the in vitro Anthelmintic investigation, we created a suspension of our crude extract at a dosage of 200 mg/ml with chitosan HCl as a thickening agent, and Pyrantel Pamoate was utilized as a standard. Phertima posthuma was employed in Anthelmintic research due to its morphological and physiological similarity to the roundworms found in the colon. The Omakai seeds were saved for extraction, and the heat maceration process was used. We obtained a 99% yield from the pure water (powder) sample, 82% from the 70:30 HA sample, and 0% from the pure isopropyl alcohol sample. Phytochemical screening followed normal procedures. Our crude extract contained alkaloids, tannins, saponins, glycoside reducing sugars, and flavonoids, but no anthraquinone. The sample of pure water (powder) exhibits the least period of paralysis and death, and when compared to the reference standard Pyrantel Pamoate, there is only a difference of 12 seconds for paralysis and 20 seconds for worm death. We concluded that our assessment was successful since the average time of paralysis and death for extract of pure water (powder) was the shortest when compared to other extracts, and when compared to Pyrantel Pamoate, there was a difference of 12 seconds for paralysis and 20 seconds for earthworm death.
INTRODUCTION:
Helminth is a Greek word that means "worm." It has a plague history. The helminth egg was discovered in mummified human feces over 10,000 years ago, and we recognized several clinical symptoms of helminth infection in Hippocrates' writings, Egyptian medical papyri, and the Bible.1 Helminths have a low death rate, but their morbidity is high enough to raise the burden of noncommunicable diseases in a number of developing nations. Helminth infections are prevalent in children2.
They are divided into two groups: flatworms (Platyhelminthes) and nematodes (roundworms). Roundworms have a strong cuticle, but flatworms have a plasma membrane. All helminths have quite big body structures and are classified as active feeders. Most flatworms are hermaphroditic or bisexual, whereas roundworms have different sexes. The exposure of helminthic illnesses relies on the weather, food choices, cleanliness, and interaction with vectors.3 Nematodes, particularly males, have supplementary sex organs in addition to the alimentary canal and breathing system. However, the tapeworm does not have an alimentary canal, therefore it gets nutrition from the tegument.3-4 Roundworms and flatworms are intestinal helminths that cause infection in humans. The larvae stage occurs in an intermediate host, whereas the adult worms live in a mammalian host. Other nematodes, such as hookworms and whipworms, do not have a pulmonary phase in their life cycle. There is no need for an intermediate host: eggs travel through the intestinal lumen, and owing to fecal contamination of food, infected eggs are consumed, and a new cycle of infection begins; for example, a naturally occurring Heligosomoides polygyrus is restricted to the host's gut.5-8 As new synthetic anthelmintic drugs or vaccines are unlikely to become available in the near future, alternative parasite infection control strategies are urgently required. Several medicinal plants (anthelmintics) have been used to treat gastrointestinal nematode and other helminth infections.9-11 Vernonia amygdalina and Annona senegalensis are two plants commonly employed by Nigerian farmers and live stalk handlers. In a recent study, Omakai was discovered to have an anthelmintic impact on Ascaridia galli and other parasites. It is the plant that has the greatest proteolytic enzymes and has been utilized since ancient times12. Therefore, the present investigation was carried out on Omakai to evaluate the Anthelmintic property.
MATERIALS AND METHODS:
Isopropyl alcohol, Misopropyl alcohol, Acetone, saline solution, Drangendroff's reagent, 10% ammonia solution, Felhing's reagent, 1% gelatin solution, and sodium chloride solution were obtained from Rankem Pvt. Ltd Mumbai of the local market, distilled water from the scientific shop of the local market, and Pyrantel Pamoate from a local medical store.
Plant Material Collection and Authentication:
Omakai seeds were utilized in the evaluation. These seeds were harvested from ripe, fresh Omakai purchased from the Amravati local market and validated by the College of Agriculture in Shri Shivaji Horticulture College (281/2024). The seeds were then rinsed with a saline solution to remove any undesirable debris, and let to dry at room temperature. After drying, these materials went through extraction (maceration).13-15
Processing of Plant Material:
The Omakai seeds were rinsed with saline water to remove dirt and foreign contaminants. Following that, the harvested seeds were air dried for 7 to 8 days. After drying, the seeds were weighed and put in a clean, airtight zip lock pouch.16
Extraction Process:
Following drying, the seed test was put something aside for extraction. The extraction was performed utilizing the hot maceration process, according to standard strategy. To start with, the examples were isolated into seven gatherings for extraction; each contained a 10g test. The first was with unadulterated water, the second was with unadulterated water containing seed powder (for this situation, we shaped coarse seed powder), the third was with unadulterated isopropyl alcohol (99.9%), and the hydroalcoholic arrangements for the excess examples were ready in various focus proportions.
The first was 70:30 HA, which contained 70ml of liquor and 30ml of water; the second was 30:70 HA, which contained 30ml of water and 70ml of liquor; the third was 50:50 HA, which contained 50ml of water and 50 ml of liquor; and the last one was made with isopropyl alcohol, misopropyl alcohol, water, and CH3)2CO. Each of the seven examples were joined for heat maceration for 24 hours, following which the filtrate was gathered and put away for dissipation. After dissipation, the strong powdered was gotten as an item.11-21
Collection of Test Organisms (Earthworms) and Authentication:
The earthworms of the species Phertima posthuma were utilized for study according to the normal procedure 13-16 and were gathered from the damp soil of the garden of Shri Shivaji College of Horticulture, Amravati, and validated at the Shri Shivaji College of Agriculture, Amravati. Because of their morphological and physiological similarities to roundworms found in the gut, earthworms were utilized to study anthelmintic activity.22-23
Phytochemical Analysis of Extract:
The solid powder extract produced following extraction was utilized for phytochemical screening. The phytochemical screening was carried out for secondary metabolites such as alkaloids, tannins, saponins, reducing sugar for glycosides, flavonoids, and anthraquinone glycosides according to the protocols stated in standard literature.23-25
In -Vitro Anthelmintic Study:
To investigate the Anthelmintic activity of the obtained extract, a suspension of the crude extracts was produced at a dosage of 200mg/ml with chitosan HCl as a thickening agent. As a typical medication, we utilized a 200mg/ml Pyrantel Pamoate solution. For in vitro Anthelmintic testing, we followed normal protocol with few variations.25-30. Prior to beginning the experimental method, all extract suspensions were newly produced in distilled water. Seven groups were created for in vitro anthelmintic activity testing. Each group was given three petri dishes with the same medication dose and one earthworm (Phertima postuma), as well as Pyrantel Pamoate. To investigate the Anthelmintic activity of the obtained extract, a suspension of the crude extract was produced at a dosage of 200mg/ml with chitosan HCl as a thickener.
As a typical medication, we utilized a 200mg/ml Pyrantel Pamoate solution. Complete observations were taken until the paralysis and death of a particular worm, which can be found in each Petri dish. The times of paralysis and death of those worms were also recorded.31-32
RESULTS AND DISCUSSION:
Extraction Yield:
The extraction yield of all Omakai seed extracts in different solvents was different, because we extracted in different solvents in different concentration ratios, such as 70:30 HA, 30:70 HA, 50:50 HA, pure water (seeds), pure water (Seed Powder), pure isopropyl alcohol, and multi-solvent (25% isopropyl alcohol, 25% misopropyl alcohol, 25% water, 25% acetone). The extraction yield of all the various solvents was measured in grams and translated to a percentage. We obtained 0% yield from the pure isopropyl alcohol sample, but 99% and 82% yield from the 70:30 HA, pure water (seeds) sample, respectively.
Table 1: Extraction Yield of different solvent extracts of Omakai
|
Extracts |
Percentage Yield |
|
Pure water (seeds) |
82% |
|
Pure water (Powder) |
66% |
|
Pure isopropyl alcohol |
0% |
|
70:30 HA |
99% |
|
30:70 HA |
49% |
|
50:50 HA |
58% |
|
(Multi solvent Water, acetone, isopropyl alcohol, misopropyl alcohol. (25:25:25:25) |
60% |
Phytochemical Screening:
The results of phytochemical screening are shown in Table 2. We did phytochemical screening using normal protocols and detected the existence of various secondary metabolites such alkaloids, tannins, saponins, reducing sugars of glycosides, and flavonoids in our plant material; nevertheless, we received a negative result for anthraquinone glycosides.
In Vitro Anthelmintic Study:
In an in vitro Anthelmintic investigation, the average period of paralysis and death varied according on the concentration ratio of the solvent extracts used. The results of this in vitro Anthelmintic investigation are shown in Table 3. Figures 1 exhibit a graphical depiction of the in vitro Anthelmintic results. The standard or positive control was a suspension of Pyrantel Pamoate at a dosage of 200mg, whereas the negative control was the usual saline solution.
Table 2: Result of Phytochemical Analysis of Different Solvent Extracts of Omakai
|
Extracts |
Alkaloids |
Tannins |
Saponins |
Anthraquinone |
Reducing Sugars of glycosides |
Flavonoids |
|
Pure water (seeds) |
+ |
+ |
+ |
- |
+ |
+ |
|
Pure water (Powder) |
+ |
+ |
+ |
- |
+ |
+ |
|
70:30 HA |
+ |
+ |
+ |
- |
+ |
+ |
|
30:70 HA |
+ |
+ |
+ |
- |
+ |
+ |
|
50:50 HA |
+ |
+ |
+ |
- |
+ |
+ |
|
Multi solvent |
+ |
+ |
+ |
- |
+ |
+ |
[(+) Present, (-) Absent]
Table 3: Average time of Paralysis and Death of Phertima posthuma against various extracts and Reference Drugs
|
Concentration Ratio (200mg/ml) |
Result |
|||
|
Time of Paralysis (mins) |
Mean (mins) |
Time of death (mins) |
Mean (mins) |
|
|
Control |
- |
- |
- |
- |
|
Pure water (seeds) |
5.04 5.45 1.45 |
3.98 |
1.00 6.50 3.17 |
3.55 |
|
Pure water (Powder) |
3.58 5.38 3.50 |
3.48 |
1.10 1.48 2.50 |
1.70 |
|
70:30 HA |
4.00 4.10 4.15 |
4.08 |
3.12 3.03 3.20 |
3.11 |
|
30:70 HA |
6.27 7.40 5.35 |
6.34 |
2.35 3.43 3.18 |
3.00 |
|
50:50 HA |
3.42 3.53 5.57 |
3.50 |
2.48 2.45 2.45 |
1.46 |
|
Multi Solvent (Water, isopropyl alcohol, acetone, isopropyl alcohol) (25:25:25:25) |
11.00 10.55 11.00 |
10.08 |
11.54 10.37 12.03 |
11.31 |
|
Pyrantel Pamoate (Positive control) |
3.35 3.37 3.36 |
3.36 |
0.52 1.10 1.10 |
0.90 sec or 1.50 |
Figure 1: Evaluation Parameters of Various Extracts
CONCLUSION:
For our experiment, we started by gathering seeds from ripe fresh Omakai in Amravati's local market. Following harvesting, these seeds were treated using a saline solution to eliminate dirt and other pollutants. After washing, the seeds were left to dry for 7 to 8 days. After drying, the seeds were extracted. The extraction was carried out utilizing heat maceration. Following the extraction yield, we acquired the extraction yield of pure water (seeds), and the sample of 70:30 HA demonstrated significant amounts of extraction yield, which were 99% and 82%, respectively, in contrast to other samples, while a sample of pure isopropyl alcohol yielded 0%.
Following that, the phytochemical analysis of our obtained extract was performed, with all tests carried out according to standard procedure and revealing the presence of tannins, alkaloids, saponins, flavonoids, reducing sugar for glycosides, and negative results with the anthraquinone test. Then, an in vitro Anthelmintic research was conducted to assess anthelmintic activity. For that reason, we gathered earthworms of species Phertima posthuma from the damp soil of the garden and then conducted an in vitro study on earthworms. In the in vitro Anthelmintic study, the average time for paralysis of all extracts, i.e., pure water (seeds), pure water (powder), 70:30 HA, 30:30 HA, 50:50 HA, and 50:50 HA, and Pyrantel Pamoate, was 3.98, 3.48, 4.08, 6.34, 3.50, 10.08 (mins), and for death 3.55, 1.70, 3.11, 3.00, 1.46, 11.31, and 1.50 (mins).
However, pure water (powder) takes the least amount of time for both death and paralysis, with a difference of only 12 and 20 seconds when compared to the reference standard Pyrantel Pamoate. As a result of the foregoing investigation, we found that Omakai seeds' Anthelmintic activity was satisfactorily tested.
REFERENCES:
1. Malviya VR, Pande SD, Bobade NN. Preparation and evaluation of sustained release beads of zolmitriptan hydrochloride. Research Journal of Pharmacy and Technology. 2019; 12(12):5972-6.
2. Malviya V. Preparation and evaluation of emulsomes as a drug delivery system for bifonazole. Indian Journal of Pharmaceutical Education and Research. 2021 Jan 1; 55(1):86-94.
3. Kousalya M, Geetha P, Jesuraja A, Kumar MV. In-Vitro study of anthelmintic activity of Eclipta prostrata (L) various Extracts. Research Journal of Pharmacy and Technology. 2017; 10(1):58-60.
4. Babu ND, Kodithala S, Murali R, Srinivasan N. Anthelmintic activity of misopropyl alcoholic bark extract of Buchanania axillaris (Desr.). Research Journal of Pharmacy and Technology. 2018; 11(4):1298-300.
5. Malviya V, Ladhake V, Gajbiye K, Satao J, Tawar M. Design and characterization of phase transition system of zolmitriptan hydrochloride for nasal drug delivery system. International Journal of Pharmaceutical Sciences and Nanotechnology (IJPSN). 2020 May 31; 13(3):4942-51.
6. Burange PJ, Tawar MG, Bairagi RA, Malviya VR, Sahu VK, Shewatkar SN, Sawarkar RA, Mamurkar RR. Synthesis of silver nanoparticles by using Aloe vera and Thuja orientalis leaves extract and their biological activity: a comprehensive review. Bulletin of the National Research Centre. 2021 Dec; 45:1-3.
7. Malviya V, Thakur Y, Gudadhe SS, Tawar M. Formulation and evaluation of natural gum based fast dissolving tablet of Meclizine hydrochloride by using 3 factorial design 2. Asian Journal of Pharmacy and Pharmacology. 2020; 6(2):94-100.
8. Shagana S, Navenaa S, Vijay J, Vishali S, Rajendran V. Investigation of in vitro Anthelmintic activity of Ocimum basilicum Linn.(Lamiaceae). Research Journal of Pharmacy and Technology. 2021; 14(1):52-4.
9. Malviya VR, Tawar MG. Preparation and evaluation of oral dispersible strips of teneligliptin hydrobromide for treatment of diabetes mellitus. International Journal of Pharmaceutical Sciences and Nanotechnology (IJPSN). 2020 Jan 31; 13(1):4745-52.
10. Malviya VR, Pande SD. Road CKN. Preparation ad Evaluation of Zolmitriptan Hydrochloride Lozenge. J Pharma Res. 2019; 8(8):624-9.
11. Malviya V, Manekar S. Design, development and evaluation of aceclofenac and curcumin agglomerates by Crystallo Co-Agglomeration technique. Research Journal of Pharmacy and Technology. 2021; 14(3):1535-41.
12. Eswaraiah MC, Reddy DP, Illendula S. In-vitro anthelmintic activity of Tamilnadia uliginosa fruits. Research Journal of Pharmacy and Technology. 2019; 12(1):321-3.
13. Sawarkar HA, Khadabadi SS, Muley NB, Aswar PB. Anthelmintic Activity of Extracts of Trigonella Foenum-Graecum Linn. Research Journal of Pharmacy and Technology. 2009; 2(1):128-30.
14. Sutar NG, Giri MA, Kendre PN, Syed NL, Singh J. Phytochemical Investigation and Pharmaological Screening of Alienthus excelsa Linn. Leaves as Anthelmentic activity in Earthworm. Research Journal of Pharmacy and Technology. 2010; 3(1):244-6.
15. Malviya V, Pande S. Development and Evaluation of Fast dissolving Film of Fluoxetine hydrochloride. Research Journal of Pharmacy and Technology. 2021; 14(10):5345-50.
16. Malviya V. Design and Characterization of Thermosensitive Mucoadhesive Nasal Gel for Meclizine Hydrochloride: Thermosensitive Nala Gel of Meclizine HCL. International Journal of Pharmaceutical Sciences and Nanotechnology (IJPSN). 2022 Feb 28; 15(1):5782-93.
17. Tehseen F, Ghori SS, Dawood M, Tazeen S, Sara K, Samad A. Phytochemical screening and invitro anthelmintic activity of a novel polyherbal formulation. Research Journal of Pharmacy and Technology. 2021; 14(5):2742-4.
18. Mishra A, Pradhan DK, Mishra MR, Behera R, Panda AK, Vaishnaw SK. In-vitro Anthelmintic Activity of Musa sapientum Linn. Flower. Research Journal of Pharmacy and Technology. 2011; 4(1): 89-91.
19. Rajesh R, Selvakumar K. Evaluation of in vitro Anthelmintic activity of Lannea coromandelica (Houtt.) Merr. against Pheretima posthuma and Ascardia galli. Research Journal of Pharmacy and Technology. 2022; 15(6):2539-42.
20. Malviya V, Burange P, Thakur Y, Tawar M. Enhancement of solubility and dissolution rate of atazanavir sulfate by nanocrystallization. Indian Journal of Pharmaceutical Education and Research. 2021 Jul 1; 55(3):S672-80.
21. Malviya V, Tawar M, Burange P, Jodh R. A Brief Review on Resveratrol. Asian Journal of Research in Pharmaceutical Sciences. 2022 May 25; 12(2):157-62.
22. Malviya V, Arya A, Burange P, Gajbhiye K, Rathod G, Tawar M. To evaluate the cardioprotective effect of hydroalcoholic extract of Matricaria chamomilla linn in isoproterenol induced myocardial infarction in wistar rats. Research Journal of Pharmacy and Technology. 2022; 15(9):3887-92.
23. Polireddy P, Malviya V, Arora S, Singh M, pooja Tanaji G, Devhare LD, Ruikar DB, Dharmamoorthy G. Assessment of Hepatoprotective Potential of Ecbolium Linneanum Extract on Experimental Animals. Journal of Coastal Life Medicine. 2023 May 29; 11:884-90.
24. Malviya V, Tawar M, Burange P, Bairagi R. Preparation and Characterization of Gastroreten-tive Sustained Release In-situ Gel of Lafutidine. International Journal of Pharmaceutical Sciences and Nanotechnology (IJPSN). 2022 Dec 12; 15(6):6216-28.
25. Malviya V. Investigation of in-vitro antidiabetic study, antioxidant activity and Anthelmintic property of various extracts of bitter cumin seeds: antidiabetic study, antioxidant activity, and Anthelmintic property of bitter cumin seeds. International Journal of Pharmaceutical Sciences and Nanotechnology (IJPSN). 2023 Jul 31; 16(4): 6855-74.
26. Chiranjeevi U, Kamarajan K, Manna PK. Investigation of in-vitro anthelmintic activity of Lindernia madayiparense. Research Journal of Pharmacy and Technology. 2018; 11(1):183-6.
27. Malviya V, Pande S. Design and Characterization of pH Dependent Phase Transition System of Almotriptan Malate for Nasal Drug Delivery System by Employing Factorial Design. Ind. J. Pharm. Edu. Res. 2024 Jan 1; 58(1): 76-90.
28. Gayathiri NM, Sudhakar P, Manimekalai P, Sabarinath C. In Vitro Anti-helminthic Activity of hydroalcoholic extract of Martynia annua L. and Pentanema indicum. Research Journal of Pharmacy and Technology. 2019; 12(8): 3847-50.
29. Mate M, Kulkarni AS, Mehta PP, Dhamane SP. Evaluation of anthelmintic activity of Cassia tora L. leaves against Pheretima posthuma. Research Journal of Pharmacy and Technology. 2013; 6(10):1152-3.
30. Malviya V. Box-Behnken Modeling Served for the Development and Optimization of Nanoparticles Loaded with Perindopril and Erbumine. Asian Journal of Pharmaceutics (AJP). 2023 Sep 15; 17(3).
31. Manju V, Revathi R, Murugesan M. In vitro antioxidant, antimicrobial, anti-inflammatory, anthelmintic activity and phytochemical analysis of Indian medicinal spices. Research Journal of Pharmacy and Technology. 2011; 4(4): 596-9.
32. Satao J, Wadaskar P, Bais A, Bhamburkar S, Malviya V. Preparation and Characterization of Benidipine Hydrochloride Microballoons. International Journal of Scientific Development and Research. 2020; 5(6): 176-81.
|
Received on 11.10.2024 Revised on 13.01.2025 Accepted on 25.02.2025 Published on 03.05.2025 Available online from May 05, 2025 Asian J. Pharm. Res. 2025; 15(2):104-108. DOI: 10.52711/2231-5691.2025.00017 ©Asian Pharma Press All Right Reserved
|
|
|
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License. |
|