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Journal of Drug Delivery and Therapeutics

Open Access to Pharmaceutical and Medical Research

Copyright  © 2024 The  Author(s): This is an open-access article distributed under the terms of the CC BY-NC 4.0 which permits unrestricted use, distribution, and reproduction in any medium for non-commercial use provided the original author and source are credited

Open Access   Full Text Article                                                                                                                                            Research Article

Formulation And Evaluation of Herbal Suspension by Using Factorial Design

Ijajahamad S. Rukadikar ¹*, Amol A. Dixit ², Dnyaneshwari T. Raykar ³, Rutuja R. Sakpal ⁴, Prasad S. Repal ⁵ 

Department of Pharmaceutics, Sarojini College of Pharmacy, Rajendranagar, Kolhapur- 416004 Maharashtra, India. 

 

 

INTRODUCTION

Herbal remedies play a crucial role in developing effective therapeutic agents and have demonstrated potential in combating various ailments. Research in herbal medicine faces challenges in identifying active compounds from plant sources. Authentication through research, whether on whole herbs or extracted compounds, is crucial. Herb-herb and herb-drug interactions, especially in polypharmacy, require increased awareness and study. This approach can lead to the re-evaluation and redesign of agents for enhanced efficacy and fewer side effects. The current imperative is to develop safer drugs for various disorders, driving interest in the pharmacological evaluation of plants used in traditional medicine systems ¹. Factorial design, a statistical research methodology, analyses how various variables interact within an experiment. In a full factorial experiment, multiple factors with discrete levels are considered, and all possible combinations of these levels across the factors are tested. This design, also known as a fully crossed design, allows researchers to explore the individual effects of components as well as their interactions on the response variable. Typically, factorial experiments involving only two levels, resulting in treatment combinations such as in a 2x2 factorial design, which examines four treatment combinations in total ².

MATERIAL AND METHOD

Materials

API :-

Synonym: golden apple, bael. Biological name is Agele Marmelos belonging to family Rutaceae. It contains phytochemicals such as coumarin, marmelos, ageline etc. It possess anti-inflammatory, anticoagulant property³.

Synonym: Tamal patri, tej patta. Biological name is Cinammomum tamala belonging to family Lauraceae. . It contains phytochemicals such as, procyanidin and cinnamaldehyde. It possess anti-hyperglycaemic property⁴.

Excipients:-

It is used as suspending agent, stabilizing agent.

It is used as emulsifying agent.

 It is used as preservative.

It is used as sweeting agent.

It is used as flavouring agent.

Method

Bay leaves and Bael leaves were obtained from the local nursery market in the Kolhapur region. All chemicals utilized were of analytical grade. The leaves were subsequently dried in the shade, grounded into a fine powder, and employed in the preparation of a suspension. Bay & Bael powder was passed through sieves size 100 mesh size individually. Both the powder drugs were mixed by trituration using mortar and pestle. Subsequently, powder mixture was passed through sieve size 100 mesh size. Further, water was added into the powdered drug mixture. Finally, sodium CMC, Tween 20 were added, followed by sodium benzoate, aspartame and lemon oil were added, along with final volume make upto 100 ml with water. The preparation was filled into amber colored plastic bottle and labelled it¹. 2² factorial design was used for identifying the optimum formulation

Factors- Factor A (conc. CMC), Factor B (conc. Tween20)

Levels- Low and high concentration².

Table 1: Factors and levels:

 

 

Table 2: Factor and combinations:

 

Table 3: Formulation table:

Sr. No.	Ingredients	Batch 01	Batch02	Batch03	Batch04
1	Bael Powder	1g	1g	1g	1g
2	Bay Powder	1g	1g	1g	1g
3	Sodium CMC	1%	2%	1%	2%
4	Tween 20	0.5%	1%	1%	1%
5	Sodium Benzoate	1g	1g	1g	1g
6	Aspartame	0.1g	0.1g	0.1g	0.1g
7	Lemon oil	1ml	1ml	1ml	1ml
8	Purified Water	q. s	q. s	q. s	q. s

 

 

Evaluation

 Morphological parameter of bael and bay leaves powder like Colour, odour, taste were evaluated.

1. Phytochemical evaluation⁵
2. Alkaloid Test: 

The extract was mixed with diluted HCl and Mayer’s reagent to detect alkaloids.

The extract was combined with 5 ml of distilled water and Benedict’s reagent to identify carbohydrates.

1. Saponin Test: 1 ml of the extract was mixed with 20 ml of distilled water and shaken for 15 minutes to test for saponins.
2. Phenol Test: 

The extract was treated with 10% lead acetate to detect phenols.

The extract was combined with 10% lead acetate to determine the presence of tannins.

Few drops of NaOH were added to the extract, followed by HCl, to identify flavonoids.

a) Sedimentation Volume: 

The sedimentation volume is the ratio of the ultimate height (Hu) of the sediment to the initial height (Ho) of the total suspension as the suspension settles in a cylinder under standard condition⁷.

b) Redispersibility: 

The suspension was allowed to settle in measuring cylinder. The mouth of the cylinder was closed and was interred through 1800 and the number of inversion necessary to restore a homogeneous suspension was determined⁷. 

c) Rheology 

The time required for suspension sample to flow through a pipette was determined the apparent viscosity of suspension and can be calculated using the following equation⁷.

 Flow rate = volume of pipette (ml)/ Flow time 

d) Viscosity 

The viscosity of the sample was determined at room temperature using Brook field viscometer⁶.

 e) pH

The pH of suspension was determined using pH meter⁶.

 f) Crystal growth

 Stability of suspension will also reduce because of crystal growth, which usually occurs from temperature variation during storage and form broad particle size distribution. Crystal formulation was determined at room temperature (RT) ⁶.  

RESULT AND DISCUSSION

The morphological tests revealed distinct characteristics for Bay and Bael powders. Bay powder exhibited a greenish-brown color, an aromatic odor, and a slightly spicy taste. In contrast, Bael powder displayed a yellowish-brown color, was odorless, and had a slightly bitter taste. These differences highlight the unique properties of each herb, which are essential for their identification and potential therapeutic applications. Understanding these sensory attributes is crucial for ensuring the correct use of each herb in polyherbal formulations. Results are mentioned in table 6.

 

 

Table 6: Morphological Test

 

 

The phytochemical evaluation of Bay and Bael extracts indicated the presence of alkaloids, saponins, phenols, tannins, and flavonoids, while carbohydrates were absent. These compounds are crucial for the formulation's therapeutic efficacy and stability. Alkaloids offer anti-inflammatory and antimicrobial benefits. Saponins enhance bioavailability and possess antimicrobial and immune-boosting properties. Phenols provide antioxidant protection, improving formulation stability. Tannins aid in wound healing and exhibit anti-inflammatory effects. Flavonoids contribute antioxidant and anti-inflammatory benefits. Understanding these phytochemicals ensures that the formulation is both effective and stable, supporting its therapeutic claims and overall efficacy Results are mentioned in table 7.

 

 

Table 7: Phytochemical Evaluation

Sr. No.	Test	Observation		Inference
		Bay	Bael	Bay	Bael
1.	Alkaloid: Extract + dil. HCl + Mayer’s reagent	Yellowish or orange insoluble pigment	Yellowish or orange insoluble pigment	Alkaloid present	Alkaloid present
2.	Carbohydrate: Extract + 5ml distilled water + Benedict’s reagent	No ppt	No ppt	Carbohydrate  absent	Carbohydrate  absent
3.	Saponin: 1ml Extract + 20ml distilled water + Shake for 15 minutes	Formation of Foam	Formation of Foam	Saponin  present	Saponin  present
4.	Phenol: Extract + 10% lead acetate	White ppt	White ppt	Phenol  present	Phenol  present
5.	Tannin: Extract + 10% Lead acetate	White ppt	White ppt	Tannin  present	Tannin  present
6.	Flavonoid: Extract +Few drops on NaOH + Few drops of HCL	Yellow color disappears on addition of HCl	Yellow color disappears on addition of HCl	Flavonoid  present	Flavonoid  present

 

 

Herbal suspension was prepared, and stability parameters were evaluated.

Physical Evaluation

Pharmaceutical formulations, such as suspensions, often require preservatives, coloring, flavouring agents, and other additives. It is essential to demonstrate the necessity of adding a preservative at the appropriate level, as well as its physical and chemical compatibility with other ingredients in the medicinal product. Various excipients were used to enhance the stability of formulation. Sodium CMC improved viscosity and stability. Tween 20 was as a surfactant / suspending agent Sodium Benzoate was used as a preservative. Lemon oil and aspartame were used as flavouring and sweeting agent respectively. Physical evaluation parameters are enlisted in table 8.

 

 

Table 8: Physical Evaluation Parameters 

Sr. No.	Parameter	Batch 01	Batch 02	Batch 03	Batch 04
1.	Nature	Liquid	Liquid	Liquid	Liquid
2.	Colour	Greenish	Greenish	Greenish	Greenish
3.	Odor	Pleasant	Pleasant	Pleasant	Pleasant
4.	Taste	Sweet	Sweet	Sweet	Sweet

 

 

Formulation study

Formulation studies for all the batches were performed. From amongst all the batches, batch 02 demonstrated a sedimentation volume of 1.3, signifying a stable suspension; a flow rate of 5 ml/26.60 sec, indicating smooth fluidity; and a viscosity of 61.4 cp, which strikes an ideal balance between thickness and ease of handling. The pH remained consistently neutral at 7 ± 0.57, and no crystal growth was observed, suggesting good long-term stability. The factorial design clearly showed that the high concentrations of sodium CMC and Tween 20 used in Batch 02 resulted in the most effective formulation properties. Hence, batch 02 was found to be optimum.

 

 

Table 9: Formulation study

Sr. No.	Parameter	Batch 01	Batch 02	Batch 03	Batch 04
1.	Sedimentation volume	1.9	1.3	0.8	0.64
2.	Flow Rate	5ml/4.30sec	5ml/26.60sec	5ml/50.40sec	5ml/1.30min
3.	Viscosity	58.6 cp	61.4 cp	59.8 cp	63.3 cp
4.	pH	7+ 0.57	7+ 0.57	7+ 0.57	7+ 0.57
5.	Crystal growth	No	No	No	No

                    

 

CONCLUSION

The development and evaluation of the polyherbal suspension demonstrated that adherence to World Health Organization guidelines is essential for creating effective and stable herbal products. The study highlighted the importance of using various excipients to enhance the formulation's stability. 

Using a 2² factorial design to optimize the formulation, batch 2 emerged as the optimal combination of low and high concentrations of CMC and Tween 20. This batch exhibited excellent redispersibility, appropriate sedimentation properties, and optimal viscosity. The stability parameters and pH levels remained suitable, with no significant changes in the physicochemical and organoleptic properties of the suspension.

Overall, the study successfully demonstrated the necessity and compatibility of the selected additives and excipients, leading to a stable and effective polyherbal suspension. This approach can serve as a model for future developments in herbal product formulations.

 

 

 

Conflict of Interest: The authors declare no conflict of interest.

Authors’ Contribution: All authors contributed to the preparation of the manuscript. All authors read and approved the final manuscript.

Funding: None.

Acknowledgment: We are grateful to Guide Mr. Amol A. Dixit Head of Department Pharmaceutics Department, Sarojini College of Pharmacy, Kolhapur for providing the necessary guidance chemicals and Facilities for completing the project work.

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