INTRODUCTION:

Indian system of medicine has a deep root in our cultural heritage and caters to the Medicare of large section of our population. This system mainly used herbs, recent times there has been a marketed shift towards herbal cures because of pronounced cumulative and irreversible ill effects of many modern drugs. Indian system of medicine (Ayurveda, Siddha, Unani, Yoga and Naturopathy) has developed over a long period of time. Herbal medicine is also called botanical medicine or phytomedicine. Herbalism is a traditional medicine practice based on the use of: a plant seeds, berries, roots, bark or flowers and plant extract for medicinal purposes. Advances in clinical research show the value of herbal medicine in the treating and preventing disease. Plants had been used for medicinal purposes long before recorded history.

Okra (Abelmoschus esculentus L.) is a flowering plant of the Malvacae family which is also known as lady’s finger, gumbo, bamya or bamia or commonly known as bhindi. Natural polymers have been used in different pharmaceutical formulations. They are easily available, non-toxic, biodegradable and cost effective to be used as pharmaceutical excipients. In present investigation we have reviewed about method for extraction and characterization of mucilage (Hibiscus esculentus) and further characterized to be used as a pharmaceutical excipient. Main focus of review was to study about anti-cancer activity of okra mucilage. Different methods for isolation and physicochemical method for characterization was focused. Antioxidant activity as well as IR spectra determination was noted. Okra is rich in phenolic compounds with important biological properties like quartering and flavonol derivatives, catechin oligomers and hydroxycinnamic derivatives. Okra is also known for being high in antioxidants activity. Okra has several potential health beneficial effects on some of the important human diseases like cardiovascular disease, type 2 diabetes, digestive diseases and some cancers.

Nowadays, plant gums and mucilage have evoked large interest due to their various pharmaceutical applications as diluents, binders, disintegrants thickeners and gelling agents in gel making them attractive substitutes for costly synthetic excipients. Binders play important role in tablet formulation as tablet formulation to impact cohesion on powder mix and hence improve on the flow properties of the granules. A majority of the investigations on natural excipients in drug delivery systems have centered on proteins and polysaccharides, due to their ability to produce a wide range of materials and properties according to molecular structural alteration mucilage’s are very often used in various industry. Vast application and plant mucilage gums in various industries is because of low cost and ready availability and important properties which they confer on product. Two plants are prominently used, and have been chosen for this investigation; these are the leaves of Hibiscus rosasinensis Linn and fruits of Okra (abelmoschus esculentus Linn).

This plant was formerly name as Hibiscus esculentus and was originated from India. Size of okra can be divided into three categories; large (L), Medium (M) and Small (S) with respect to their lengths. Mucilage can be termed as plant hydrocolloid having polymer of a monosaccharide. But polysaccharide is having mucilage having hydrophilic substance with high molecular weight molecules. Their swelling properties are observed by characterized by the entrapment of large amount of water between the polymer chains and branches.

From literature it was found that okra possess anticancer activity. For the detection of activity okra was isolated and characterized for significant cell growth inhibition in human breast cancer (MCF7) cells up to 63% is reported. Hot buffer extraction of okra pod could induce apoptosis in melanoma cells by interacting with Gal-3 and cause a cell cycle arrest in G2/M phase. Moreover, the fraction of okra polysaccharide extract had inhibition effects on MCF-7, Hela, and MCG803 cells with the lowest survival rates of 63.90, 63.5, 1 and 67.71%, respectively. From okra contains significant levels of protein, carbohydrate, neutral sugars, minerals and other complex polysaccharides and medically reported to be linked with anticancer, antimicrobial, hypoglycaemic, anti-ulcer activities. The rheological behavior of okra mucilage is pseudo plastic and can be used to enhance viscosity hence it acts as an ideal substitute for costly synthetic and semi-synthetic excipients.

MATERIAL AND METHODS:

Material: Okra was obtained from local market of Kolhapur, other all the ingredients used were of analytical grade obtained from unique laboratory Kolhapur.

Methods:

Extraction of mucilage:

In present study we have used two different methods for extraction of okra which can give maximum yield with minimum requirements.

First method: Step 1:

Extraction of mucilage: Hibiscus esculents fruit were used for isolation of mucilage, Firstly fruit were washed with water to clean it from dirt if any and grinded into a mixer. The material obtained was soaked in warm water for 4 hours, boiled for 2 hour and kept aside for 2 hours for release of mucilage into water. After a period of 2 hours material was squeezed in a muslin bag to remove the mark from the filtrate.

Fig.1 Okra mucilage in ethanol

Step 2: Isolation of mucilage:

Equal volume of ethyl alcohol was added to filtrate to precipitate the mucilage. The mucilage was separated, dried in oven at about 45°C, powdered and passed through sieve #80. The powdered mucilage was stored in a desiccator until further use

Fig.2 Extracted okra mucilage

Second method:

Step 1: Extraction of mucilage: Okra pods (without seeds) were sliced and immersed in water at room temperature. After 12^th with the aid of muslin cloth, the solid was separated from the liquid fraction. Three volumes of ethanol were added to the filtrate and liquid was slowly stirred by handling until mucilage was precipitated.

Step 2: Drying of sample:

The mucilage was dried for 12^th hour at 30⁰ C in an oven. Pulverized to fine powder with the aid of grinder and passed through sieves. The resultant fine powder was stored in an amber recipient until the moment of use.

Formulation of Tablet:

All ingredients were weighed and mixed properly using mortal pestle, the binder solution was mixed with the powder mixture to form an adhesive mass which can be granulated. The powder blend was screened using #22 mesh screen and to prepare weight granules. Then moist granules were dried in a hot air oven at controlled temperature not exceeding 55⁰ C. The dried granules were passed through a screen of smaller size than that used to the moist granules that are sieve no 10. After dry screening the dried and screened granules are separated into coarse and fine granules by shaking. The mixed granules were compressed in a single punch tablet press fitted with the appropriate punches.

Fig.3 Punching of tablet

Formulation Table:

Table No 1: Formulation of tablet

Batch	F1	F2	F3	F4
Drug (mg)	100	100	100	100
Binder	2.5	5	7.5	10
Magnesium stearate	12	12	12	12
Lactose	85	85	85	85
Talc	1	1	1	1

Evaluation test for binder:

Fourier Transform Infrared spectroscopy (FTIR): This technique is used to obtain infrared spectrum of absorption, emission and photoconductivity of solid, liquid and gas. The FTIR uses interferometry to record information about a material placed in the IR beam. The Fourier Transform results are spectra that analysts can us to identify or quantify the material.

Evaluation test for granules:

Angle of Repose –For determination of angle of repose a funnel with a wide outlet is fixed at a distance of 2 cm above the table, where a piece of paper is placed directly beneath the funnel. Powder is added while the funnel is closed. The contents flow through a collect on the paper. The diameter of ^{the cone (D) and two opposite sides (l}1^+l2^{)
are measured with rulers.}

Formula - tanθ = h/r.

Fig.5 Angle of Repose

Bulk density –Pass the quantity of powder in graduated cylinder must be done gently to avoid changing the nature of the material. Into a dry graduated cylinder of 250 mL gently introduce, without compacting, approximately 20 gm of the test sample (m) weighed with 0.1 percent accuracy. Carefully level the powder without compacting if necessary. Read the unsettled apparent volume to the nearest graduated limit. Calculate the bulk density in gm/mL.

Fig. 6 Bulk density

Tap density –The tapped density is obtained by mechanically tapping a graduated measuring cylinder or vessel containing the powder sample. After observing the initial powder volume or mass, the measuring cylinder or vessel is mechanically tapped and volume or mass reading are taken until little further volume or mass change is observed. The mechanical tapping is achieved by raising the cylinder or vessel and allowing it to drop, under its own mass.

Formula – m/Vt

Fig. 7 Tapped density

Evaluation tests for tablet:

Friability test –Take a sample of 10 whole tablets. Dedust the tablets carefully and weigh accurately the required number of tablets. Place the tablets in the drum and rotate it 100 times remove the tablets, remove any loose dust from them and weigh them accurately. The test is run only once unless the results are difficult to interpret or if the weight loss is greater than the targeted value, in which case, the test is repeated twice and the mean of the three tests is determined.

Formula - ^𝑤¹⁻^𝑤²× 100

𝑤1

Disintegration test -The assembly is suspended in the liquid medium in a suitable vessel, preferably a 1 liter beaker. Unless otherwise stated in the individual monograph, introduce one tablet into each tube and, if directed in the general monograph, add disc to each tube. Suspend the assembly in the beaker containing the specified liquid and operate the apparatus for the specified time. Remove the assembly from the liquid. The disintegrating tablets pass the test if all of them have disintegrated. If 1 or 2 tablets fail to disintegrate, repeat the test on 12 additional tablets.

Fig. 8 Disintegration test

Dissolution test –Place the stated volume of the dissolution medium, free from dissolved air, into the vessel of the apparatus. Assemble the apparatus and warm the dissolution medium to 36.5⁰ to 37.5⁰. Place one dosage unit in the apparatus, taking care of exclude air bubbles from the surface of the dosage unit. When apparatus is used allow the tablet to sink to the bottom of the vessel prior to the rotation of the paddle. A suitable device such as a wire of glass helix may be used to keep horizontal at the bottom of the vessel tablets that would at the beginning of each test. Lower the basket into position before rotation. Operate the apparatus immediately at the speed of rotation specified in the individual monograph. Within the time interval specified, withdraw a specimen from a zone midway between the surface of the dissolution medium and the top of the rotating basket, not less than 10 mm from the wall of vessel. Filter the sample solution promptly through a membrane filter disc with an average pore diameter not greater than 1.0 micron. Discard the first few ml of the filtrate. Repeat the whole operation five times. Where two or more tablets are directed to be placed together in the apparatus, carry out six replicate tests. For each of the tablet tested, calculate the amount of dissolved active ingredient in solution as a percentage of the stated amount where two or more tablets are placed together, determine for each test the amount of active ingredient in solution per tablet and calculate as a percentage of the stated amount.

RESULT AND DISCUSSION:

Binder:

Fourier Transform infrared spectroscopy (FTIR) -

FTIR spectrum of okra mucilage in the resolution range of 4000-400 cm^-1. In the larger band in the region of 3600 cm^-1 to 3100 cm^-1 with a sharp peak centred at 3721 cm^-1 characteristic O-H stretching vibration and hydrogen bond of the hydroxyl group. This group serves as active site for attachment of colloidal particles and okra powder.

Absorption peak at 2926.31 to 2861.60 cm^-1 correspond to C-H stretching vibration from methyl and methylene group in cellulose and hemicellulose components while 1730 cm^-1 refers to carbonyl increase C=O stretching vibration of conjugated ketone, carboxylic acid and ester in lignin and hemicellulose. The shoulder at 1460 cm^-1 may be due to presence of water in the fibers. A little peak at 1249.37 cm^-1 is associated to the angular deformation on the O-H bonding plane. The two peaks observed at 1334.05 to 1076.93 cm^-1 in the spectrum indicate blending vibration C-H and C-O groups of the aromatic rings in polysaccharide.

Furthermore the absorption peaks around 1200 cm^-1 to 904.74 cm^-1 indicate the presence of C-o bond associated with presence of functional group such as alcohol, ether and ester.

Granules:

Angle of repose –

Formula - tanθ = h/r.

Height (h) – 2 cm

Table no 2: Angle of Repose

Batch	F1	F2	F3	F4
Diameter	3.625 cm	4.364 cm	4.982 cm	5.925 cm
Radius	1.8125 cm	2.182 cm	2.491 cm	2.9625 cm
Result	47.81	42.50	38.76	34.02

Among the 4 batches of granules. Conclude that an angle of repose of F4 batch was 34.02 is better than the other 3 batches.

Bulk Density –

Formula - m/v

Table no 3 Bulk Density

Batch	F1	F2	F3	F4
Weight	7.98 gm.	7.86 gm.	8.35 gm.	8.55 gm.
Volume	18 ml	17.4 ml	19.2 ml	20 ml
Result	0.44	0.43	0.43	0.42

Among the four batches, we conclude bulk density of 4 batches. The bulk density of 4^th batch is 0.42 and it was better than other 3 batches.

Tap Density -

Formula – m/Vt

Table no: 4 Tap Density

Batch	F1	F2	F3	F4
Weight	7.98 gm	7.86 gm	8.35 gm	8.55 gm
Volume	15 ml	15.6 ml	16.3 ml	16 ml
Result	0.53	0.50	0.51	0.53

Among the four batches, we checked the tap density of 4 batches of granules and the 4^th batch

0.53 is better than other 3 batches of granules.

Tablet:

Friability test –

Formula - ^𝑤¹⁻^𝑤²× 100

𝑤1

Table no 5 Friability test

Batch	F1	F2	F3	F4
Initial weight (w1)	2.00gm	2.03gm	2.05gm	2.08gm
Final weight (w2)	1.89gm	1.92gm	2 gm	2.06gm
Result	5.5%	5.41%	2.43%	0.97%

Among the all four batches of tablets, conclude that 4^th batch is better friability than other 3 batches of tablets.

Disintegration and Dissolution test:

Table no 6 Disintegartion and Dissolution

Batch no.	Disintegration time	% Drug release
F1	64	80.30
F2	72	85.60
F3	78	95.60
F4	65	96.30

Among all the batches of tablets, conclude that 4^th batch of tablet has better disintegration time and dissolution rate than other three batches of tablets.

CONCLUSION:

The mucilage of okra was successfully isolated and extracted from the dry okra fruit, the isolation procedure was done with distilled water with ratio 1:3. Finally obtained pure water-soluble mucilage the supernant was washed with the help of ethanol. It was done because the water insoluble material was completely removed from mucilage and to obtain a pure water-soluble precipitate. After drying, okra mucilage was used in wet granulation and direct compression method in the different formulation of paracetamol. From the okra gum used as a binding agent some tablet formulation with good hardness, friability and dissolution and disintegration rate. However, this natural binder dissolution rate of some slightly soluble drug and may be consider as a good candidate for binding the formulation.

Form the study it was clear the binding property of fruit polymer of abelmoschus esculentus (okra) is much better. Natural gum are promising biodegradable polymeric materials. It is a clear that gum and mucilage have much advantage over synthetic materials. The result presented here shows that the mucilage obtained from okra can be used as a binder in paracetamol tablets formulation with good physical properties. Tablet of long disintegration times were produced, hence it’s potential in binding and prepare the granule of pharmaceutical formulation.

REFERENCE:

1. Suhailah H et al, Anticancer activity of okra raw polysaccharides extracts against human liver cancer cells, Tropical Journal of Pharmaceutical Research (2019)18 (8): 1667-1672

2. Mark L. Woolfe .et al, Studies on the Mucilage Extracted from Okra Fruits (Hibiscus esculentus L.) and Baobab Leaves (Adansonia digitata L.), J. Sci. Fd Agric. (2018)1917(28),519-529.

3. Noorlaila A et al, Emulsifying properties of extracted Okra (Abelmoschus esculentus L.) mucilage of different maturity index and its application in coconut milk emulsion, International Food Research Journal, (2015)22(2): 782-787

4. Ahiakpa J.K. et al, Mucilage Content of 21 Accessions of Okra (Abelmoschus spp L.), Scientia Agricultural, (2014) 2 (2): 96-101

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7. Ameena K et al, Isolation of the mucilage from Hibiscus rosasinensis Linn. and Okra (Abelmoschus esculentus Linn.) and studies of the binding effects of the mucilage, Asian Pacific Journal of Tropical Medicine, (2010)539-543.

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Received on 14.03.2023 Modified on 29.05.2023

Asian J. Pharm. Tech. 2023; 13(3):178-182.

DOI: 10.52711/2231-5713.2023.00032