INTRODUCTION:

The most prevalent chronic inflammatory respiratory disease, asthma is characterised by symptoms such as constriction of the airways, coughing, dyspnoea, chest tightness, and recurrent wheezing episodes Approximately 300 million people worldwide are suffering by this pervasive illness.

Patients are looking for alternative asthma treatments because current therapy is not effective for treating asthma at all stages and can have some adverse effects also. Varieties of plants documented in traditional system of medicine which gives symptomatic relief and prevent exacerbation of asthma. Several plants have been found to contain a variety of pharmacological actions targeting stabilization of mast cell bronchodilatory, anti-inflammatory, anti-allergic, anti-spasmodic, immunomodulatory activities besides inhibition of some mediators such as leukotrienes, lipoxygenase, cyclooxygenase, platelet activating, phosphodiesterase and cytokine, in the management of asthma.^1-2In the present work, we had prepared herbal anti asthmatic tablet lozenges using Sarcostemma brevistigma stems, Piper longum fruits and Solanum xanthocarpum herb. These plants are mentioned in Ayurveda as anti-asthmatic herbs and also found potentially active in experimental models of bronchial asthma.^2-4Although there are several dosage forms for treating asthma in the market, such as syrups, tablets, and inhalers, new dosage forms that work locally and effectively are still needed. Lozenges are a type of solid dosage form that contains the medication in a base that has been sweetened and flavoured. Its major purpose is to extend the action of the medication and is meant to be released gradually into the mouth.

MATERIAL AND METHODS:

Plant materials:

The dried fruits of Piper longum and herb of Solanum xanthocarpum were procured from the local area of Ahmedabad. Fresh aerial parts of Sarcostemma brevistigma were collected from Jamnagar.

Preparation of plant extracts:

70% v/v ethanol was used to extract the powdered crude drugs thoroughly, and the mixture was then filtered. After evaporation of the filtrate, the percentage yield of hydro-alcoholic extract was computed.

Formulation of compressed tablet lozenges^5-6:

The granules were prepared using the wet granulation process in accordance with the provided formula (Table 1).

Table 1. Formula for one tablet lozenge (750 mg)

Ingredients	Weight of ingredients per lozenge (750mg)	Percentage of ingredients
Solanum xanthocarpum extract	75mg (25mg × 3)	10%
Sarcostemma brevistigma extract
Piper longum extract
β-cyclodextrin	150 mg	20%
Mannitol	112.5 mg	15%
Sucrose	259.8 mg	34.64%
Sucrose syrup	113.7 mg	15.16%
Methyl paraben	0.375 mg	0.05%
Propyl paraben	1.125 mg	0.05%
Menthol	7.5 mg	1%
Magnesium stearate	7.5 mg	1%
Talc	7.5 mg	1%
Coating using (6%) alcoholic Shellac solution

A precise weight of the herbal extracts, sucrose, and mannitol were combined by trituration, and the mixture was then granulated with the use of sucrose syrup as a binder. The wet mass was further sieved, and the resulting granules were dried for four to five hours at 40°C. Subsequently, 10% fines were added and the dried granules were passed through sieve again. After adding the remaining β-cyclodextrin, magnesium stearate, propyl and methyl parabens, talc, and menthol, the dried granules were combined and compressed using a rotary punching machine. The approximate weight of each tablet lozenge was 750mg.

Evaluation of the granules and lozenges^6-9:

Angle of repose, bulk density, tapped density, hausner's ratio, and Carr's index were assessed for the granules, whereas general appearance, hardness (crushing strength), friability, disintegration time, and weight variation were measured for the lozenges.

The amount of total alkaloid¹⁰ and total phenolic¹¹ in herbal extracts, mixtures, and formulations was estimated.

Stability study of formulation¹²:

Accelerated stability testing was conducted for three months and one year at a temperature of 40±2ºC/75% RH in accordance with International Conference on Harmonisation (ICH) standards. In this study, various conditions such as high temperatures, harsh light, and mechanical stress are applied to potential formulations in order to identify which formulation provides the active components with the best stability. Tests were carried out a year later to assess the lozenges and determine the total amount of phenolic and alkaloid content.

Estimation of piperine in formulation by HPTLC Method:

High performance thin layer chromatography (HPTLC) was performed on 10cm × 10cm precoated silica gel 60 F₂₅₄ plates (E. Merck, Germany). Samples were put to the plates in bands that were 6mm broad and 12.2mm apart using Camag Linomat V applicator (Muttenz, Switzerland) equipped with a 100microlitre syringe (Camag, Switzerland). In a Camag twin-trough glass chamber (10 × 10cm), linear ascending development was carried out at room temperature using mobile phase vapour (toluene, ethyl acetate, and diethyl amine, 5: 4: 1 v/v/v). Following plate drying, the Camag TLC scanner was used to scan the plate in absorption mode at 330nm using WinCATS software (version 1.4.3.6336) with slit dimensions of 6.00 × 0.45mm. In a volumetric flask, precisely weighed 5mg of piperine were dissolved in 10 ml of methanol to make a stock solution. The stock solution was diluted with methanol to prepare standard solutions for calibration; the concentrations were set so that piperine levels ranged from 250 to 1250ng. Peak area (Y axis) against concentration (X axis) was plotted to create a calibration curve.

Five tablet lozenges were weighed, ground up, and mixed with 100% alcohol. After evaporation of the sample solution, methanol was used to reconstitute the leftovers. For estimating, 5 µl of this solution was employed and the amount of piperine in the plant was determined using the peak area values of the standards and the sample.

The method was validated in terms of linearity, interday precision, intraday precision, repeatability, accuracy, specificity, limit of detection and limit of quantification. The validation of the analytical method was conducted using guidelines from the International Conference on Harmonisation.¹³

RESULT AND DISCUSSION:

The yield of hydro-alcoholic extracts of Solanum xanthocarpum, Sarcostemma brevistigma and P_{iper longum}_{were observed to be 11.36, 9.49 and 16.85%
w/w respectively.}Compared to other herbal medications, Solanum xanthocarpum extract has the greatest total alkaloid and total phenolic content (Table 2). The tablet lozenges have elliptical shape and light yellowish brown colour with smooth texture and aromatic odour. Tables 3 and 4 provide the granules and lozenges' evaluation parameters. The formulation exhibited good stability over elevated temperature and humidity condition as degradation rate of active constituents was slow during stability study (Table 5).

Table 2. Total alkaloids & total phenolic content of herbal extracts

Herbal drugs	Total alkaloid content (%w/w)	Total phenolic content (%w/w)
Solanum xanthocarpum	0.529	0.802
Sarcostemma brevistigma	0.078	0.661
Piper longum	0.244	0.592
Extract mixture	2.491	5.819

Table 3. Evaluation parameters of granules

Evalution Parameter	Result ± SD (n=3)
Bulk density (gm/ml)	0.617 ± 0.0016
Tapped density (gm/ml)	0.656 ± 0.0021
Hausner’s ratio	1.063 ± 0.0017
Carr’s index	5.942 ± 0.15
Angle of repose	28.86 ± 0.46

Piperine was detected in the extract by TLC analysis at R_f0.32, precisely matching the reference standard (Figure 1). It was observed that the formulation contained 0.012±0.006% w/w of piperine. The measured LOD and LOQ were 52.25ng/spot and 158.34ng/spot, respectively. The intraday and interday coefficient of variation for piperine varied from 0.21 to 1.15% and 0.51 to 1.53% respectively. %R.S.D. for repeatability of measurement of peak area and repeatability of sample application were found to be 0.30% and 0.77% respectively. Average % recovery for method reported was 99.98. Since the method effectively resolved the component even in the presence of other phytoconstituents in the sample, it is specific to piperine.

Table 4. Evaluation parameters of tablet lozenges

Evaluation Parameter	Result _{± Sd (N=3)}	Result (After 1 Year Stability Study)
_W_{idth (mm)}	_{9.89 ± 0.29}	_{9.89 ± 0.29}
Length _(mm)	_{15.13 ± 0.07}	_{15.13 ± 0.07}
Thickness _(mm)	_{6.13 ± 0.01}	_{6.13 ± 0.01}
Average weight _(mm)	_745.99	_745.99
Hardness test (n)	_{243.33 ± 4.16}	_{251.25 ± 3.2}
Disintrgration time (min.)	_{16.10 ± 0.51}	_{16.58 ± 0.45}
Friability (%)	_{0.159 ± 0.003}	_0.995
Total alkaloid content (%w/w)	_0.2576	_0.2415
Total phenolic content (%w/w)	0.5849	0.5743

Table 5. Stability study and estimation of total alkaloid and total phenolic content

Time	Total alkaloid content per lozenge (mg)	% Remained	Total phenolic content per lozenge (mg)	% Remained
Initial phase	1.932	100	4.387	100
1^st phase (1^st month)	1.924	99.48	4.353	99.90
2^nd phase (3^rd month)	1.916	99.17	4.342	98.97
3^rd phase (1 year)	1.903	98.55	4.325	99.26

Calibration curve of standard piperine

Densitometric chromatogram

HPTLC Chromatogram

Figure 1. HPTLC study of piperine

CONCLUSION:

Herbal anti asthmatic lozenges were formulated and evaluated for various parameters in the current investigation. The prepared lozenges were shown to be stable while stored in strips, according to a one-year physical stability testing. The formulation may be used as an alternate medication for respiratory conditions including asthma.

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Received on 26.04.2024 Revised on 17.07.2024

Accepted on 19.10.2024 Published on 18.04.2025

Available online from April 22, 2025

Asian J. Res. Pharm. Sci. 2025; 15(2):113-116.

DOI: 10.52711/2231-5659.2025.00017

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