Formulation and Evaluation of Fast Dissolving Tablets containing Clopidogrel bisulfate using holy basil seeds as a natural superdisintegrant

 

Omprasad Nayakal*, Prajakta Patil, Mangesh Bhutkar, Dheeraj Randive, Somnath Bhinge

Rajarambapu College of Pharmacy, Kasegaon, Tal- Walwa. Dist- Sangli, Maharashtra -India 415404

 

ABSTRACT:

The motto of the present research work was to develop fast dissolving tablets of Clopidogrel bisulfate by using Holy basil seeds mucilage as a natural super disintegrating agent. Clopidogrel is a thienopyridine with antiplatelet activity. It irreversibly alters the platelet receptor for adenosine diphosphate. To formulate fast dissolving tablets of Clopidogrel bisulfate direct compression technique was employed using 6,12 and 18 mg concentrations of Holy basil seed mucilage which acted as a natural super disintegrant. Also compatibility of the drug and Holy basil seed mucilage was studied by using FTIR study. The powdered Holy basil seeds were characterized for various parameters namely ash value, solubility and pH. The prepared tablets were evaluated by using various parameters. The results of weight variation test revealed that it was within the accepted limits. Thickness and hardness of the formulated tablets were between 2.6±0.25 to 3.1±0.32mm and 3.1±0.11 to 3.0±0.21 kg/cm² for the prepared batches. Wetting time of the tablets was found to be between 22±0.54 to 35±0.57sec. The results of in-vitro disintegration time of the tablets varied from 32±1.2 to 40±1 sec. for the prepared batches. Formulation F3 comprising of Holy basil seed mucilage (30mg) was also compared F6, containing 30 mg of Crospovidone as super disintegrating agent. It was observed that the drug release from formulation F6 was 91.9% in 10min, whereas formulation F3 exhibited 99% drug release at the end of 10 minutes. Thus, it was concluded that Holy basil seed mucilage possess good potential of partly replacing the commercial superdisintegrant(s).

 

 

 

INTRODUCTION:

A different solid dosage form is drug delivery system like tablets, capsules, sachets and pills as well as a bulk or unit-dose powders and granules. Amongst the various dosage forms, oral solid dosage forms have greater importance and occupy a prime role in the pharmaceutical market.^[1] Oral drug delivery system is mostly preferred for administration of various drugs.^[2]

 

Solid dosage forms are popular because of ease of administration, accurate dosage, self-medication and most importantly the patient compliance. However, the pediatric and geriatric patients face a great difficulty in swallowing of the solid dosage forms. Therefore, recent advancements in novel drug delivery systems have resulted in a convenient dosage form for administration and to achieve better patient compliance by means of fast dissolving tablets (FDTs). Fast dissolving tablets are the novel types of tablets that disintegrate /dissolve /disperse in saliva. Their characteristic advantages such as administration without water, anywhere, anytime, lead to their suitability to geriatric and pediatric patients^[3]. The different natural gums and polymers are widely used for pharmaceutical industries. Mucilages are used in different context such as superdisintegrant, binder, sustaining agent, thickening agent, suspending agent as well as gelling agent etc. Natural polymers are preferred over synthetic and semi-synthetic materials due to their low cost, free availability, non-toxic, non-irritating nature and environment friendly.^[4-5]

 

Clopidogrel Bisulfate is a thienopyridine with antiplatelet activity. Clopidogrel bisulfate irreversibly alters the platelet receptor for adenosine diphosphate (ADP), thereby blocking the binding of ADP to its receptor, inhibiting ADP-mediated activation of the glycoprotein complex GPIIb/IIIa, and inhibiting fibrinogen binding to platelets and platelet adhesion and aggregation.^[6] Clopidogrel is an inhibitor of platelet aggregation that is used to decrease the risk of myocardial infarction and stroke in patients known to have atherosclerosis.^[7] Different uses of parts of the plant e.g. leaves, flowers, stem, root, seeds etc medicinal properties have been attributed to Holy basil. They recommended for the mostly for have been used traditionally as anti-diabetic, antifertility, hepatoprotective, hypotensive, hypolipidmic, expectorant, analgesic, anticancer, antiasthmatic, antiemetic, diaphoretic and anti-stress agents. Holy basil has also been used in the treatment of fever, bronchitis, arthritis, convulsions, etc.^[8-9]

 

The mucilage is extracted from the seeds of the plant Holy basil. The seeds were available from local market. The flowering top of the plant with the seeds were send for the authentication to Botanical Survey of India. The plant was authenticated as Holy basil. The aim of the study was to evaluate the superdisintegrant property of the Holy basil seed mucilage with compared other Superdisintegrants used were crospovidone. The model drug used was Clopidogrel bisulfate Present study involves the formulation of Clopidogrel fast disintegrating tablets using the mucilage obtained from the seeds of Holy basil and evaluated.^[10]

 

MATERIALS AND METHODS:

MATERIALS:

Clopidogrel bisulfate was obtained as a gift sample from the Yarrow Chemicals Mumbai. Holy basil were purchased from local market. Microcrystalline Cellulose and Mannitol were obtained from Qualigens, Mumbai. Crospovidone was obtained as a gift sample from Cipla Ltd, Mumbai. Talc and Magnesium stearate was procured from Supreme Chemicals.

 

METHODS:

Isolation of Mucilage from Holy basil seeds:

Holy basil seeds were properly dried and were triturated in a mortar using a pestle. The powdered seeds were defatted using petroleum ether as a defatting agent. The defatted mass was soaked in distilled water for 12 h. The swollen mass was then dried in an oven at 60^oC. Thereafter it was passed through sieve no.40.The obtained mucilage was stored in a desiccator till further use.^[11]

 

Preparation of Clopidogrel Fast Dissolving Tablets:

Clopidogrel bisulfate tablets were prepared by using direct compression method. The composition of each tablet is as shown in Table No1. All the ingredients were properly mixed together. Talc and Magnesium stearate were passed through mesh 40. The powder blend was compressed into tablets on 8 station rotary punch tabletting machine and same hardness was used for the required number tablets. Compressed tablets were examined as per official standards and tests. Tablets were packaged in a well closed light resistant and moisture proof containers.^[12]

 

FTIR spectroscopy was used for compatibility study:

Drug and Holy basil seed mucilage were mixed in 1:1 ratio and spectrum of the drug, excipient mixture was scanned using FTIR spectrophotometer in a range 4000-400cm-1.^[13]

 

Table 1: Formulation of Clopidogrel FDTs by direct compression method

Ingredients	F1 (mg)	F2 (mg)	F3 (mg)	F4 (mg)	F5 (mg)	F6 (mg)
Clopidogrel bisulfate	75	75	75	75	75	75
Mucilage	6	12	18	-	-	-
Mannitol	3	3	3	3	3	3
Talc	5	5	5	5	5	5
Magnesium Stearate	1	1	1	1	1	1
MCC	110	104	98	110	104	98
Crospovidone	-	-	-	6	12	18
Total Weight (mg)	200	200	200	200	200	200

 

Characterization of dried mucilage:

The obtained mucilage was subjected to physical evaluation such as color, odor, taste etc. Other parameters assessed included ash value, pH, and solubility. Also phytochemical screening tests were performed for the detection of alkaloids, carbohydrates, glycosides, saponins, phytosterols, phenols, tannins, flavonoids, amino acids, diterpenes etc.^[14-15]

 

Pre compression evaluation: ^[16]

1.    Angle of Repose:

Angle of repose was determined using fixed funnel method. The blend was poured through a funnel that can be raised vertically until a maximum cone height (h) was obtained. Radius of the heap (r) was measured and angle of repose was calculated using formula:

 

 

θ= tan^-1(h/r)

 

2.    Bulk density:

It is the ratio of total mass of powder to the bulk volume of powder. It was measured by pouring the weight powder (passed through standard sieve # 20) into a measuring cylinder and initial volume was noted. This initial volume is called the bulk volume. From this, the bulk density is calculated according to the formula mentioned below. It is expressed in g/ml and is given by:

 

 

Bulk density	=	Weight of granule
		Volume occupied by granule

 

3.    Tapped density:

It is the ratio of total mass of the powder to the tapped volume of the powder. Volume was measured by tapping the powder for 750 times and the tapped volume was noted. It is expressed in g/ml and is given by:

 

Tapped density	=	Weight of granule
		Tapped volume occupied by granule

 

4.    Hausner’s ratio:                     

It is an indirect index of ease of powder flow. It is calculated by the following formula:

 

Hausner’s ratio	=	Tapped density
		Bulk density

 

5.    Carr’s compressibility index:

It is calculated by the following formula:

 

% Compressibility	=	Tapped density - Bulk density	X    100
		Tapped density

 

Post compression evaluation parameters: ^[17-21]

1) Hardness:

It is the force required to break a tablet in a diametric compression force. Monsanto hardness tester was used for test, which applies force to the tablet diametrically with the help of an inbuilt spring.

 

2) Friability:

The friability of a sample of 20 tablets was measured. Tablets of all batches were used to evaluate friability as per USP XXIV monograph.  The testing was done by BESTO Apparatus.

 

3) Disintegration time:

In- vitro disintegration test was carried out on 6 tablets using digital tablet disintegration test apparatus. Distilled water at 37^oC was used as a disintegration media and time in second taken for complete disintegration of tablet with no residue remaining in apparatus was noted.

 

4) Stability study:

Stability of the optimized formulation F3 was tested according to the ICH guidelines, at 40±2^oC/75%±5% RH condition in stability camber for 3 months. Tablets were tested for drug content for 30, 60, and 90 days.  

 

5) Wetting Time:

A piece of tissue paper folded twice was placed in a small petri dish (ID 6.5cm) containing 6ml of pH 6.8 buffer. A tablet was put on the paper, and the time for complete wetting was measured. Three trials for each batch were performed.

 

6) In vitro release profile of formulated Tablets:

Dissolution test of tablets was performed using buffer pH 6.8 and with USP dissolution type II apparatus at 50 rpm and 37 ± 0.5 °C temperature. Test sample (5 ml) was withdrawn at particular time interval and replaced with fresh dissolution media maintained at 37±0.5 °C. The test sample was filtered and analyzed using UV spectrophotometer at λ max 254 nm. The results of the study are presented Table 5.

 

7) Drug Content:

A quantity of powder equivalent to 2 mg of Clopidogrel bisulphate was accurately weighed and transferred into a 100 ml volumetric flask and dissolved in 0.1 N HCl and the volume was made with 0.1 N HCl (pH 1.2). The solution was then filtered through Whatmann’s filter paper. The stock solution was suitably diluted with 0.1 N HCl and the absorbance were measured at λ max 254 nm using UV / visible spectrophotometer. 

 

8) Thickness:

Thickness was determined for 20 tablets using a Digital Vernier calliper and the average thickness was determined in mm.

 

9) Weight variation test:

All the prepared tablets were evaluated for weight variation as per USP XXIV monograph. Thirty six tablets of each batch were used to evaluate weight variation among tablets and mean and standard deviation was calculated.

 

RESULTS AND DISCUSSION:

Table 2: Organoleptic evaluation of the mucilage

Parameter	Observations
Appearance	Brownish ash like hygroscopic powder
Odor	Characteristic
Taste	Mucilaginous
Ash value	12%
pH	6.1
Solubility	Swell in water

 

Preformulation Studies:

Solubility Studies:

Clopidogrel bisulfate is soluble in water, 4.5 pH acetate buffer, pH 6.8 phosphate buffer and slightly soluble in ethanol.

Compatibility study:

 

Fig 1:  FTIR studies of Holy basil seed mucilage

 

 

Fig 2:  FTIR studies of Clopidogrel bisulfate

 

 

Fig 3:  FTIR studies of Holy basil seed mucilage and Clopidogrel bisulfate

 

 

 

All the principle peaks of Clopidogrel bisulfate were retained in FTIR spectrum of 1:1 physical mixture of Clopidogrel bisulfate and Holy basil seed mucilage. Hence, there was no interaction between Clopidogrel bisulfate and the Holy basil seed mucilage. Thus, the drug and excipient are compatible with each other as show in Fig 3.

 

 

Table 3: Pre compression evaluation

Parameters	F1	F2	F3	F4	F5	F6
Bulk density (g/ml)	0.44±0.17	0.44±0.57	0.43±0.43	0.45±0.49	0.43±0.871	0.44±0.14
Tapped density (g/ml)	0.52±0.11	0.51±0.12	0.52±0.015	0.53±0.21	0.52±0.01	0.53±0.23
Compressibility index (%)	11.5±0.25	16.4±0.23	17.3±0.24	15±0.24	17.3±0.31	16.4±0.54
Hausner’s ratio	1.1±0.27	1.12±0.24	1.1±0.23	1.12±0.24	1.13±±0.23	1.1±0.21

 

All formulations showed Pre compression parameters of that they posses good properties.

 

Table 4: Post compression evaluation parameters

Parameters	F1	F2	F3	F4	F5	F6
Hardness(kg/cm2)	2.6±0.25	3.1±0.32	2.9±0.31	2.8±0.21	3.0±0.25	3.0±0.32
Thickness (mm)	3.1±0.11	3.0±0.12	2.9±0.21	3.0±0.21	3.1±0.14	3.1±0.15
Friability (%)	0.21±0.15	0.26±0.01	0.40±0.14	0.24±0.13	0.20±0.52	0.21±0.14
Disintegration time(sec)	40±1.0	34±1.1	32±1.2	54±1.5	47±1.1	42±1.1
Drug Content (%)	99.21±0.64	98.21±0.48	97.36±0.32	99.32±0.45	98.65±0.48	98.23±0.65
Wetting Time	35±0.57	25±0.42	22±0.54	52±0.52	43±0.47	37±0.55
Weight Variation (mg)	Pass	Pass	Pass	Pass	Pass	Pass

 

Table 5: Dissolution profile of Clopidogrel bisulfate fast disintegrating tablets

Time (min)	Cumulative percentage of drug release
	F1	F2	F3	F4	F5	F6
1	82.4	84.6	90.2	71.4	76.7	75.7
2	84.3	85.1	91.0	73.3	77.9	78.1
3	85.9	87.3	92.3	74.9	79.6	79.3
4	89.7	88.4	93.4	75.3	81.4	81.3
5	90.5	91.4	94.2	76.7	82.2	83.9
6	91.6	92.6	95.9	77.3	83.2	84.4
7	94.1	94.9	96.4	78.2	85.2	86.9
8	95.1	95.4	97.6	79.4	85.4	88.3
9	95.3	96.9	98.5	81.3	86.3	90.7
10	96.2	97.2	99.0	82.0	86.9	91.9

 

 

 

Fig 4:  Cumulative Percentage Release of Clopidogrel bisulfate

 

 

 

 

 

 

   

(A)                                           (B)

Fig 5: Disintegration pattern of prepared fast dissolving tablets of Clopidogrel bisulfate containing Holy basil seed mucilage

(A) At time t=0 seconds (B) At time t=10 seconds

 

All the batches of tablets were evaluated for the different parameters and the results of the same are shown in Table No.3 and Table No. 4. Weight variation for the prepared tablets was found within the specifications of USP. Hardness of all the formulations F1-F6 was found within the range of 2.6±0.25 to 3.1±0.32 kg/cm² which is within the acceptable limit. Friability of all formulations was also found to be within the acceptable limits. The friability for all formulations was found to be less than 1%. The drug content in different formulation was highly uniform and in the range 97.36±0.32 to 99.32±0.45%. It was observed that (F1- F3) batches which comprised of Holy basil seed mucilage exhibited less in-vitro disintegration time between 40±1 to 32±1.2seconds and showed the wetting time in the range of 22±0.54 to 35±0.57 seconds. The results were compared to other batches (F4-F6) which comprised of Crospovidone. In vitro Clopidogrel bisulfate release was 96.2% (F1), 97.2% (F2), and 99 % (F3) using holy basil seed mucilage and 82% (F4) 86.9% (F5) and 91.9% (F6) by using Crospovidone. Formulation F3 which comprised of 18mg of Holy basil seed mucilage was found to be the best formulation as it achieved cumulative drug release of about 99.0% within 10 minutes as compared with other batches.

 

CONCLUSION:

The results of the present study verified the superdisintegrant property of Holy basil seed mucilage. The tablets disintegrated much faster and consistently when Holy basil seed mucilage was used as a superdisintegrant as compared with Crospovidone. Thus, it can be concluded that Holy basil seed could be used as a natural superdisintegrant in the formulation of fast dissolving tablets.

 

 

 

 

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Received on 05.04.2018          Modified on 17.05.2018

Accepted on 06.07.2018     ©A&V Publications All right reserved