Development of Herbal Ayurvedic Formulation as Digestive Tablets, Evaluation of it’s Pharmaceutical, Pharmacognostic Parameters and Screening of its Antioxidant Potential

 

Vikas Sharma¹*, Chandana Majee¹, Rahul Kaushik², Shivani Saxena,¹ Salahuddin ¹,

Avijit Mazumdar¹

¹Department of Pharmaceutical Chemistry, Noida Institute of Engineering and Technology (Pharmacy Institute), Plot No. 19, Knowledge Park-2, Greater Noida, Uttar Pardesh-201306, India.

²Department of Pharmacognosy, Ram-Eesh Institute of Vocational and Technical Education, Plot No. 03, Knowledge Park-1, Greater Noida, Uttar Pradesh-201306, India.

 

ABSTRACT:

Herbal digestive tablets are meant for treating indigestion problems. The indigestion problem is one of the major problems of all (the) ages of human beings. As trends for eating fast foods is increasing, simultaneously the improper digestion also tends to increase. There are a number of digestive tablets in the market but in attempt to improve their taste the actual motto behind their use is masked. To combat the indigestion problems, in the present study an attempt has been made to formulate, develop and evaluate herbal digestive tablets. The formula of the digestive tablet has been decided after deep review of Ayurvedic formulary of India. The ingredients of this formulation have been procured from authentic sources. The wet granulation method was used to prepare the granules for punching the tablets. After preparation, the herbal digestive tablets were subjected to various pharmaceutical evaluations and quality control evaluations as per the guidelines from World Health Organization (WHO). The formulation was also subjected to antioxidant screening using Phosphomolybdenum method. The digestive tablets are obtained as light brown-colored round tablets with pleasant odour and spicy taste with an average size of 8mm and smooth edges. Maximum extractive value was observed as 34% in methanol with a total ash value of 10.16%. Other parameters reported as bitterness value- 0.69 units, volatile oil content-8%, loss on drying- 12.3%, swelling and foaming index of 0.27 and less than 100 respectively. The tablets showed a total antioxidant potential of 0.51mg/mg as Ascorbic acid equivalent. Tablets also pass various pharmaceutical evaluation parameters like hardness, friability, weight variation, and disintegration test. Herbal digestive tablets have very excellent taste due to less bitter drugs. The tablet formula can be applied to prepare large scale production of digestive tablets.

 

 

INTRODUCTION:

Indigestion is the problem held with pain in the upper abdomen, heartburn or by improper eating of foods. Indigestion can be a daily life problem for some old age groups of peoples. Herbal digestive tablets are designed for the treatment of the indigestion conditions as they contain the herbal medicaments. Herbal drugs are the popular form of traditional medicine and the high international demands due to their ease of availability, there lesser side effects.

 

These herbal formulations are the property or knowledge of the old ages of every home. Herbal drugs also have a huge holistic believes like Holi tulsi, Ficus tree etc.¹ Digestive tablets have become a popular demanding material now a days. Ayurveda shows that herbal digestive tablets can contain pomegranate seeds, cumin seeds, black pepper, dalchini, cardamom, asafoetida, salt, black pepper, sipernate, chhoti pippali, rock salt, ginger, black salt, and other excipients.

 

After careful examination of ingredients of various digestive tablets available in the market, a formula is designed for a herbal digestive tablet which along with tasty features is also able to provide digestion improvement benefits. In this study, proper formulation of herbal digestive tablets was carried out using the formula as given in the Table 1. The tablets are also evaluated for various pharmaceutical and pharmacognostic quality control parameters as per the guidelines from World Health Organization.²

 

MATERIALS AND METHODS:

Herbal digestive tablets contain the following ingredients:

Pomegranate³:

It consists of the fruits of the Punicagranatu bearing deciduous shrub which belongs to the Lythraceae type of family.  It provides helpful nutrients and supplements and nutrients to the body.

 

Dalchini⁴:

It is a spice obtained from the inner bark of several tree species Cinnamomum verum plant belonging to the family Lauraceae. People use cinnamon as a supplement to treat problems with the digestive system, diabetes, loss of appetite, and other conditions. It has also been used in traditional medicine for bronchitis.

 

Cardamom⁵:

It consists of the fruits and seeds of the Elettaria cardamomum which belongs to the family Zingiberaceae. Cardamom is known for its own properties such as a very tasty flavoring agent, basics for medical formulations, for treating improper digestion or flatulence.

 

Asafoetida⁶:

The biological source of the asafoetida is the completely dried latex or  (gum oleoresin) excluded from the rhizomes or the taproot of several species of herb Ferula foetida This spice is used as a digestive aid, in food as a condiment and in pickles.

 

Kala and safed jeera⁷ :

It consists of the dried seeds of the Cuminum cyminum which belongs to the family Umbelliferae. Cuminum cyminum contains many types of phytochemicals that have numerous activities such as antioxidant nature, carminative action and anti-flatulent activity.⁸

 

Kala Namak⁹:

It is also named as black Himalayan salt. Kala Namak is a complex of the mineral compound having the strong sulfur content. This is a very rich source of iron which helps to treat the gas/ acidity problems,flatulence and indigestion. Black Fermented Himalayan Salt is used as a laxative and digestive aid.

 

Kali Mirch^10,11:

It consists of the spice of the Piper nigrum which belongs to the Piperaceae. It is believed to have good flavoring, spicing activities with aids to the best properties of the digestion.

 

Chhoti pippali¹⁰:

It consists of the Piper longum which is said to treats the digestion problems. According to Ayurveda, it provides the best digestion power to organize health.

 

Ginger¹⁰:

ginger consists of the rhizomes of the zingiber officinalis which belongs to the Zingiberaceae family. It is believed to have good carminative and digestive properties. It also treats the bloating properties.

 

Amla¹²: 

It is also known as the Indian gooseberry. It consists of the fruits of the embellica officinalis. It provides the nutrition and antioxidant properties to the formulation. Amla contains tannins and a very good rich source of antioxidants.

 

Other Ingredients:

Complete Formula is depicted in table no.1

 

Table 1: Ingredients used for manufacturing of herbal digestive tablets and their probable use

 

METHOD:

Tablets were prepared using the wet granulation method of tablet formation.  Granules were prepared for the proper flowing of the powderous material. Tablets are being formulated by compressing the powdered drug or the granules using the tablet punching machine. Tablets are punched in the tablet punching machine by pressing granules in die with the help of lower and upper punch. Tablet formation takes place by the combined pressing action of two punches of the punching machine i.e.  (lower and upper) and a die. Then, the pharmaceutical and pharmacognostic evaluations were performed as per the Indian pharmacopeia and WHO guidelines respectively.

 

PHARMACEUTICAL EVALUATION OF DRUGS:

Following evaluation parameters were followed for the quality control determination of the herbal digestive tablets:

 

Direct observation test:

Using the direct observation test we can determine the morphological characters of the prepared tablets. We can observe color, odor, taste, size, shape and the actual diameter of the prepared tablets. The Organoleptic observations of the tablets were presented in Table 2.

 

Friability:

Tablets in a specifies amount kept on a ROCHE FRIABILATOR and the number of tablets crushed was weighed. We can define friability as the percentage of weight that is lost by testing tablets due to the powered/mechanical action during the test in Roche friabillator. then, the Tablets are pre-weighed and after weighed then friability can be expressed as a % loss on pre-test tablets weight. For herbal digestive tablets, it was found to be 0.465 %.

 

Hardness:

Hardness is defined as the force required breaking a tablet from on its diameter. Hardness is important for testing the strength of the tablets. The hardness test of the tablets evaluation helps us to check the strength of the tablet material, binder etc. It is measured using the Monsanto tester and Pfizer.  

 

Disintegration test:

The disintegration test of digestive tablets was performed as per USP where the disintegration apparatus fitted with 6 glass tubes with a 10 number mesh at the bottom, each tube is 3 inch long was used. This arrangement of 6 tubes is placed in a distilled water which is maintained at 37^oC±2^oC, in 1 liter vessel. This system is made to move up and down through a distance of 5 to 6 cm at a frequency of 28 to 32 cycles per minute. DT was found to be 12 minutes.

 

Weight variation test:

As per Indian Pharmacopeia 1996; we took a group of 20 tablets and went for the weighing balance. Content uniformity is an important tool in the formation of quality full tablets. Sample passed the test.

 

PHARMACOGNOSTIC STANDARDIZATION:

Obviously, herbal tablets are formulated using herbs and herbs relates to the soil, environment etc. but we have to produce the best quality medicine also by that we can export our medications to the international market. So, quality control parameters as per the WHO guidelines² were performed using the following instruments:

 

1. Macroscopic characters²:

The herbal digestive tablets formulation was observed for color, odor, taste, size, shape, touch and fracture^.13 These are the characteristics that can be measured using direct visualization. The observations and the outcomes for the morphological characters were recorded in Figure 1 and Table: 2.

 

Figure:1 Visual Characterization of developed Tablets

 

2. Microscopic characters²:

This method is used for the identification of drugs on the cellular level. It is used to determine the structure of organized drugs by their histological characters. It includes the whole, certain parts of powdered crude drugs.¹⁴ These tablets were crushed, finely powdered and then used for the identification of the microscopical characters. The observations (after the treatment with the drops of water, safranine and glycerol) have shown the presence of cork cells, Ca oxalate crystals, lignified cells and fibres.

 

3. Foreign matter²:

Foreign matter is the extra matter or exogenous substance in our product. Herbal matters should be free from the soil parts, animal wastes or exudates, etc. this matter can be determined using the visual inspection method, sieving method and the magnifying lance detection method. In visual inspection, we short out the matter by viewing it by direct naked eyes. 6X and 140X lances are used to detect foreign matter of drugs. In the sieving method; 250 no. sieve is used to sort out the foreign matter^.15 Weigh the portions of this isolated/ sorted foreign matter should be within the range of  0.05 g. The calculated content of each group in grams per 100 g of air-dried sample.

 

4. Extractive value²:

The powdered herbal digestive tablets were subjected to the hot and successive. Extractive values are helpful to have an idea about the solubility, chemical moiety or chemical composition of the herbal drugs. We also can determine the solubility criteria of herbal drugs. Powdered herbal material of the drug (4g) was packed in the heating apparatus i.e. Soxhlet apparatus separately for each type of solvent like petroleum ether (60⁰-80⁰ C), chloroform, methanol, hydro alcohol (50:50) and water and extraction was carried out for 6 hr. Each extract was evaporated to dryness at their respective boiling points and constant extractive values were determined. Results are depicted in table 2.

 

5. Ash value²:

Ash value indicates the total inorganic compound or matter present in the drug. This was determined using the muffle furnace.

 

5.1. Total ash²:

Around two to four gm of herbal plant material was weighed and it is drawn to a pre-ignited silica crucible. The weighed material is spread in the bottom of silica/ platinum crucible and kept for ignition in muffle furnace at 550-620 °C temperature^.16 After 3 to 4 hours switch off the apparatus and let it cool down to the room temperature. The use of desiccators for the cooling of crucibles can also be applied.  The ash should be completely white in color that indicates the carbon-free material. The collected ash is taken, weighed and the amount of total ash is calculated.

 

5.2. Acid insoluble ash²:

Ash obtained from the silica crucible is weighed and subjected to the determination of AIA. Ash was boiled with 25ml dilute HCl (6N) for five minutes. The insoluble matter collected on an ashless filter paper, and washed with hot water for the removal of acidic residues and ignited at a temperature not exceeding 450^oC to a constant weight and the data was recorded.

 

5.3. Water-soluble ash²:

The collected ash is taken and it is dissolved in the specific amount of distilled water. Then, filter the above mixture and the insoluble ash is collected on ashless paper. Then, ignite it on 470 C to a constant weight. Now water-soluble ash can be calculated by a formula and recorded in table no. 2.

 

Table 2: Standardization parameters and observed values

 

6. Fluorescence analysis²:

Analysis of the drug-using fluorescence activity is as reported procedure. Initially, powder was observed as such in daylight and then under UV light and its high and short wavelengths and then subsequently, the powder sample was treated with distilled water, 1N NaOH in water, 1N NaOH in methanol, 50% Nitric acid, 50% HCl, conc. H₂SO₄, acetone, conc. HCl and chloroform and other chemicals. The resultant colors were observed under ultraviolet light and have shown color variations.

 

7. Phytochemical evaluation^2-3

The phytochemical evaluation includes the testing of all the chemicals of phyto origin to check their class whether they are alkaloids or glycosides etc. The petroleum ether (60°-80°C), chloroform, acetone, methanol and water extracts of the plant material were subjected to primary phytochemical screening or testing for the identification or detection of phytocomponents and shown in table no. 3.¹⁷

 

Table 3: Phytochemical screening of various extracts of the digestive tablets

*PE: Petroleum ether; C: Chloroform; EA: Ethyl acetate; M: Methanol; A: Aqueous; HA: Hydroalcohol (50:50%)

 

8. Loss on drying²:

Method:

2g of air-dried material (uniformly crushed tablets) was placed in a previously weighed and dried petri dish. The sample was dried in an oven at 100°-105°C for 3 hours and weighed. It was dried kept in Hot air oven at 100°-105°C for 3 hrs and weighed. It was again kept at the same temperature and weighing was repeated at an interval of 1 hr. 2 consecutive weighing readings not differing by more than 5mg. The loss of weight (moisture loss) in mg/g of plant dried material was calculated and the data was recorded.

 

9. Determination of bitterness value²:

Preparation of Standard solutions:

10µg/ml stock solution of Quinine HCl was prepared in drinking water. 9 different dilutions were prepared.¹⁸

 

Stock and diluted quinine hydrochloride solutions:

The stock solution of the test drug was prepared by dissolving 1g powdered drug in 100ml drinking water. 9 different dilutions were prepared.

 

Method followed:

Rinse the mouth with safe and clean, drinking water, 10ml of the maximum diluted solution was tasted for bitterness for 30 secs. In case of a delayed sensation waiting period of 1 min. was followed. After a gap of 10 min. the next higher concentration was tested in a similar manner. Threshold bitter concentration is the minimum concentration on which a diluted sample starts to provide a bitter sensation after 25- 30 seconds.

 

10. Determination of swelling index²:

The swelling index is the volume in ml taken up by the swelling of 1g of plant material under specified conditions. Plant material will get swell only when it contains the matter which can absorb water like mucilage etc.¹⁹  Its determination is based on the addition of water for plant material. By the Use of a 25ml glass measuring cylinder, the content was shaken continuously for 1 hour and then allow it to stand for a required period of time. The volume of the mixture (in ml) was then read and recorded in Table 2.

 

11. Determination of foaming index²:

Preparation of decoction:

1g of accurately weighed coarse powder was transferred into a 500ml conical flask containing 100ml boiling water and maintained at moderate boiling for 30 minutes. The resulting solution was cooled and filtered into a 100ml volumetric flask and sufficient water was added to make up the final volume.

 

The decoction was poured into a 10 stoppered test tubes (height 16cm, diameter 16mm) in successive portions ranging from 1ml-10ml and final volume was adjusted to 10 ml with distilled water^.[13] Tubs were stoppered and shaken lengthwise for 15 sec (2 shakes/sec). Further, the tubes were kept without disturbance for 15 min. and the height of foam was measured. The results have given in table no.2.

 

12. pH determination²:

the pH of the following drug solutions was determined by using previously calibrated pH meter and recorded in common table 3.

a.     1% w/v solution of drug in water

b.     10% w/v solution of drug in water

 

13. Antioxidant activity²⁰:

Phosphomolybdenum Assay- Total Antioxidant Activity

The total antioxidant activity of methanolic extract of herbal digestive tablets was calculated spectrophotometrically by Phosphomolybdenum assay using the method given by Prieto et al, 1999; Singleton VL and Rossi JA, 1965 with slight changes.¹⁴ The assay is based upon the reduction of Mo(IV) to Mo(V) by the extract or standard drug and subsequent formation of a green phosphate Mo(V) complex at acidic pH.

 

Procedure:

1.     0.3 ml of methanolic extract of herbal digestive tablets powder in the concentrations of 50-500 mg/ml in methanol was mixed with 2.7ml of Phosphomolybdenum reagent which is a combination of 28mM Sodium phosphate and 4mM Ammonium molybdate in 0.6 M Sulphuric acid) in capped test tubes.

2.     The reaction mixture was then incubated on a water bath for 100 minutes at 95 0 C. After incubation, allow the test tubes to cool at room temperature and the absorbance of the dilutions was measured using a UV spectrophotometer at 695 nm.

3.     The Blank contains 0.3ml methanol in place of drug along with phosphomolybdenum reagent.

4.     The percentage Total Antioxidant Capacity (TAC) can be calculated using formula and mentioned in the table 4.

5.     Make a standard curve. y = absorbance and x = concentrations of Ascorbic acid in mmol/mL. Find the regression line equation.

6.     Express the total antioxidant capacity as mmol equivalents of Ascorbic acid per gram of dried extract.

7.     Make a standard curve. y = absorbance and x = concentrations of Ascorbic acid in µg/mL. Find the regression line equation in figure 2.

8.     Calculate the mean and standard deviation of the parallels.

9.     TAC was expressed as Ascorbic Acid Equivalent (AAE) in µg/mg.

 

Figure: 2 : Total antioxidant potency of formulation is recorded in graphical representation in figure 6.

 

14. Micro-organism determination²:

Herbal digestive tablets were powdered and extracted and then, the suspension like matter is obtained which contains 1 ml of plant material which will be examined.  Now, it is properly mixed and incubated at 35-38°C for 2 days. Then, subcultures are prepared on a plate containing agar media and again incubated at 35-38°C for 2 days. If there is not any growth; of microorganisms is under detection, then the material qualifies the test. If there is the growth of colonies of Gram-negative rods, viewing it with a greenish color. The next following method may also be used.  Place 1 to 3 drops of a new fresh prepared 0.01 g/ml solution of N, N', N'- tetramethyl-phenylene diamine dihydrochloride R on filter-paper and apply it on a suspected colony;  the test will be positive if a purple color is obtained within 4-10 seconds.  The plant material qualifies the test if the cultures of the same do not appear.  The results for the given test is recorded in table 2.

 

15. Thin layer chromatography²:

Thin-layer chromatography is a widely used method of the pharmaceutical sector that is used for the identification, separation and qualification of the pharmaceutical/chemicals. In the TLC process two mediums are used that are named as stationary phase and mobile phase.A specific type of finely grounded silica gel or alumina is taken. It is well dispersed in water to form a slurry and this slurry is used to prepare the slides or stationary phase. It should be properly spread as a very thin layer having less than 0.25 mm thickness^.21 Additionally, the specific type of binders is also mixed with silica to provide its adherence to sticking over the glass slide e.g. silica-G. this G- indicates for the gypsum. In so many cases, a fluorescent powder is also mixed in the stationary phase to simplify the visualization in UV Fluorescence. The mobile phase consists of the chemicals or a group of chemicals that are used to run the TLC plates. The constituents of the sample run above side by the flow of the mobile phase.  We used methanol, water as aqueous phase, chloroform toluene, petroleum ether as the solvent. we also used the mixtures of the above chemical reagents. The results of the TLC are mentioned below in the respected table 4.

 

Table 4: Rf values of various TLC chromatograms using different extracts of digestive tablets.

 

16. Volatile oil determination²:

Volatile oils are called with their name due to their volatile nature. Many times, they can be termed as aromatic oils that indicate the specific aroma in the volatile oils. Volatile oils can be characterized by their odor, oily appearance and their ability to volatile at higher or room temperature. Volatile oil determination can be carried out by the steam distillation method. The distillate is collected in a pre-graduated tube and the aqueous phase is allowed to recirculation in the distillation assembly.²² The volatile oil was extracted using the Clevenger apparatus. Our herbal digestive tablets contain the light oil so light oil Clevenger apparatus was used. in the first step, 25 gm of the drug was boiled in 220 ml of water and 30 ml of glycerol with the addition of porshulin chips. It was boiled for 3.5 hours with the regular inspections. The temperature of the outgoing water was maintained. It should not be more than the 25-degree C. after 3.5 hours assembly was settled off and then 2.01 ml of oil was collected. ²³

 

RESULT:

Herbal Digestive Tablets was prepared as per the World Health Organization. Pharmacognostic evaluation Organoleptic Characteristics are recorded in Table 2 and presented in figure 1. Results of determinations of foreign matter, extractive value, ash value, water soluble ash, acid insoluble ash, loss on drying, bitterness value, swelling index, foaming index, pH, total antioxidant potency, microorganism determination and volatile oil determination are recorded in common table 2. Correlation of total antioxidant potency is correlated with ascorbic acid shown in figure 2. Records of phytochemical screening are maintained in table 3. Thin layer chromatography outcomes are depicted in table 4.

 

DISCUSSION:

The result from extractive values shows that Digestive tablets powder was having a maximum extractive value of 34% in Water and a minimum extractive value of 18% in Petroleum ether indicating a large number of phytoconstituents in the aqueous extract. A high level of ash values such as total ash 10.16%, acid depicts the increased amount of inorganic matter is present in formulation. A low moisture level was observed in Digestive tablets powder as a loss on drying 12.3% which helps to prevent the deterioration of the drugs. The microscopic study revealed the presence of parenchymatous cells, stone cells and calcium oxalate crystals in the tablet powder under the aqueous, glycerol and dil. Safranine phase. The foaming index was found to be less than 100 whereas the swelling index of 0.31 was observed in the tablets powder. Being less bitter drug shows a bitterness value of 0.69 units and a pH of 3.8 (10%). As the microbial contaminants the colony-forming unit was found to be 0.3 CFU/ml. The volatile oil was found to be light volatile oil and its quantity was 2.01 ml. The antioxidant potential of the formulation was found to be 0.51 mg/mg with reference to the ascorbic acid. The better antioxidant power inhibits the free radicals in the body which helps to be healthy by lowering the stress with an increasing aid towards better digestion.

 

CONCLUSION:

The various Pharmacognostic evaluation, physicochemical, and phytochemical standards thus obtained from this study will help in establishing the identity, purity, quality, safety, and efficacy of herbal digestive tablets. Results and the discussion forum represent that the formulation contains a good amount of inorganic content, low moisture content, aqueous soluble contents and more antioxidant potency tells that the formulation helps to improve the digestive and also provide the antioxidant effect. The antioxidants can help to improve the immune system of the body which helps to fight against many diseases. If one takes much oily foods or the increased content of free radicals then this formulation may improve the digestion of the human body as well as decrease the free oxidants of the body. The standards prepared by us can be used by many pharmaceutical industries or laboratories involved in research practice, manufacturing and the production of the herbal formulations or herbal digestive tablets to control/ manage the efficacy and quality of the herbal products; which helps in proper maintenance of the batch-to-batch consistency by which maximum therapeutic efficacy of product can be achieved.

 

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Accepted on 09.12.2020           © RJPT All right reserved

Research J. Pharm. and Tech 2021; 14(11):5849-5855.