INTRODUCTION:

Herbs and minerals are an integral part of the traditional system of medicine in many countries. Unani formulations containing metals are produced after various purification processes of metals like soaking, quenching titration, incineration, etc. Hence metals in the finished products do not possess toxicity. The compounds produced by the transmutation of base metal into noble ones along with the use of plant extracts are meant to eradicate the toxic effects of metals¹. Ayurvedic and Unani Tibb are the two most popular systems of medicine in this region where herbs, mineral products, and animal tissues are used in various dosage forms. Kushta is one of the dosage forms prepared in a very special manner, which is a blend of mineral (s) and herbal extract (s). These are claimed to be very effective but serious attempts have not been made to study and standardize this preparation on scientific parameters due to the lack of attention of the scientific community towards its standardization². WHO says standardization is a measurement for ensuring quality and is used to describe all measures that are taken during the manufacturing process and quality control leading to reproducible quality. Standardization is not an easy task as numerous factors influence their bio-efficacy and reproducible therapeutic effect³.

The selected drug for the study is Kushta-e-Baiza-e-Murgh prepared by using egg shells. Eggshell waste is fundamentally composed of calcium carbonate and has the potential to be used as raw material in the production of lime⁴. Recently researchers have utilized chicken egg shells as biofilter with a conducting polymer to enhance its sensing properties. Typically egg shells were used as bio fillers in poly aniline matrix to detect ammonia gas. The optimum ratio between eggshells and polyaniline could enhance their sensor measurement⁵. Eggshell is more than just a useful container for the nutritious yolk and egg white inside it. It is a prime example of nature’s packaging. These eggshells are beneficial for us too, Consisting of 95% Calcium carbonate (CaCO₃) making them very similar to our bones and teeth. It comes as no surprise that eggshells help to strengthen our bones and protect our tooth enamel. The shells are an amazing source of calcium just half of them provide you with your daily recommended calcium intake. It makes practical sense not only to feed the shells to your egg-laying chickens but you to utilize their unusual strength too⁶. Therefore, we can say that egg is principally a nutritious food. It is attributed to various medicinal properties in Unani medicine. Kushta-e-Baiza-e-Murghis widely used for the treatment of spermatorrhoea and Leucorrhoea. It was prepared in the laboratory by following the formula composition as mentioned in the National formulary of Unani Medicine⁷, Part-1, Government of India, Ministry of Health and Family Welfare (Department of Ayush), New Delhi, named as KBM(L) while two other commercial samples of Kushta-e-Baiza-e-MurghKBM(C-1) and KBM(C-2) were taken from local market Delhi and comparative physicochemical study has been presented.

Formula composition of Kushta-e-Baiza-e-Murgh:

The drug was prepared in the laboratory as per the formula given below in Table No. 1.

Table No. 1: Ingredients of Kushta-e-Baiza-e-Murgh

S. No.	Ingredients	Quantity
1	Post-e-Baiza-e-MurghMusaffa	100 gm
2.	Aab-e-Lemu	Q. S.

MATERIALS AND METHODS:

Kushta-e-Baiza-e-Murgh was prepared in the laboratory by following the Standards Operative Procedures (SOPs) guidelines, named KBM-L, and adapted the steps as given below.

Cleaning:

Hen’s eggs were purchased from the local market and boiled. These were cooled off and the shell was separated (See Figure 1)

Fig. 1

These separated shells of chicken eggs (Post-e-Baiza-e-Murgh) were cleaned by the process of Tasfiyah-Adviyah. In this method, the egg shells were crushed /disintegrated into small pieces and washed with lime and saline water (Namak ka solution) till the inner membrane was removed. The small pieces were again washed with clean water and dried⁸

Fig. 2

Method of preparation: The cleaned material was transferred into a china dish dipped (two-thirds) in lemon juice and taken out after the juice was absorbed with the material. The process was repeated three times as per SanatulTaklees⁹. Kushta was prepared as per SanatulTaklees Bayaz-e-Kabir with minor modifications that instead of using the cow dung cakes it was prepared in a muffle furnace because of ease of preparation and better temperature control. The peak temperature was 810^OC for 30 + 5 minutes, above 700^OC was maintained for 20+5 minutes and 600^OC was maintained for 40 + 5 minutes. After completing the process of heating, wait to cool down the whole content. The material was taken out from the furnace, triturated separately in mortar and pestle, and the finished product i.e. Kushta-e-Baiza-e-Murgh (fig. 3). It was preserved in an air-tight clean bottle.

Fig. 3.

Physico-chemical Parameters:

The parameters that were carried out in the prepared kushta are organoleptic properties, preliminary test, bulk density, tapped density, Hausner’s ratio, Carr’s compressibility index, pH values, loss in weight on drying, Total ash, Acid insoluble ash, water soluble ash, distilled water, acid-soluble extractives and loss in weight on drying at 105⁰C. Some qualitative and quantitative estimations of calcium and chloride were also performed. The tests were made in triplicate to report the mean values.

Preliminary Tests/General Tests: The following basic tests were carried out which have been performed by ancient physicians in Unani/Ayurveda systems of medicine for a long time¹⁰.

(a) Floating test: Kushta should float on the surface if it is sprinkled in a small quantity on the surface water.

(b) Fineness Test: Kushta should enter in the lines of fingers if a small quantity is rubbed between them.Fig. (4)

Fig. 4

(c) Loss of metallic luster: No metallic luster should be observed if visually examined in the presence of sunlight.

(d) Loss of metallic state: No traces of the sample should permanently stick on a silver sheet indicating no alloy formation has been taken after heating a very thin silver sheet to red for about five minutes and then allowed to cool. A metallic state can be checked by Maul Hayat.

(e) Wall Stick Test: Ideal kushta should stick upon throwing on a wall.

(f) Fuming Test: When it is put on fire it should not produce fumes.

(g) Tastelessness: No Taste should be observed in a Kushta.

Physicochemical evaluation: During the analysis, the following parameters were tested:

Bulk density and Tapped density¹¹

2gm of KBM was placed in a 10ml measuring cylinder and initial volume was noted. Then it was tapped and the tapping was continued until no further change in volume was observed. Bulk density and Tapped density were calculated and obtained in the following way.

Bulk Density= weight of the powder/Volume of the packing

Tapped density= weight of the powder/Tapping Volume of the packing

Hausner’s ratio¹²: This was calculated by the following formula.

Hausner’s ratio = Tapped Density/Bulk Density

Carr’s Compressibility index¹³: It was calculated from the equation as shown below.

Carr’s index = Tapped density – Bulk Density/Tapped Density x 100

Sedimentation Time: For the determination of sedimentation a 1.0 percent suspension was placed in a 10ml measuring cylinder and the time taken for complete sedimentation was noted.

Chemical analysis:

Determination of pH values and loss of weight on drying¹⁴

pH of 1% and 10% aqueous solution: 1% and of 10% Solution of a drug should be prepared in distilled water and filtered with the Whatman paper and pH can be measured with a digital pH meter.

Loss of weight on drying: One gram of a drug should be taken, and spread uniformly and thinly in a shallow Petri dish. It should be heated at a regulated temperature of 105^oC, cooled in the desiccators, and weighed. The process should be repeated many times till two consecutive weights are found constant. The percentage loss in weight can be calculated with respect to the drug.

Ash values^15-16

Total ash: About 1gm of drug, should be weighed accurately, in a previously ignited and tarred silica crucible. The material should be spread in an even layer and ignited by gradually increasing the heat to 500-600 °C until it is white, indicating the absence of carbon. Ash should be cooled in desiccators and weighed. The content of total ash can be calculated in terms of the air-dried drug.

Acid insoluble ash: The total ash should be boiled with 25ml of dilute hydrochloric acid for 5 minutes. The solvent should be filtered through the ashless filter paper and the insoluble matter collected on the filter paper was washed with hot water and ignited to a constant weight. The residue should be allowed to cool in the desiccators for 30 minutes and weighed without delay. The percentage of acid-insoluble ash can be calculated with reference to the air-dried drug.

Water soluble ash: The total ash should be boiled with 25 ml of distilled water for 5 minutes. The insoluble matter should be collected on an ash-less filter paper, washed with hot water, and ignited. The weight of insoluble ash should be subtracted from the weight of the total ash, giving the weight of the water-soluble ash. The percentage of water-soluble ash can be calculated with reference to air-dried drugs.

Determination of extractive values in water and acid¹⁷:500mg of a drug should be accurately weighed in a glass stopper conical flask. 100ml of water/acid should be added and it should be shaken well and allowed to stand for 1hour. 25ml of the filtrate should be transferred to a tarred flat-bottom dish and evaporated to dryness in a water bath then dried at 105°C for 6hours. Then it should be cooled in the desiccators for 30 minutes and then weigh the soluble matter present in the drug.

Estimation of calcium¹⁸

Weigh about 0.5gm of the drug, suspend it in about 100 ml water in a beaker, the heat nearly to boiling, then with stirring add sufficient hydrochloric acid to dissolve the salt. Add 2gm of Ammonium oxalate dissolved in 150 ml of water and further add the ammonia solution (1:1) dropwise with stirring, until a slight excess is present (Methyl indicator). Keep it warm with continuous stirring. A granular precipitate is obtained. Now allow the mixture to cool and decant the supernatant liquid through an ashless filter paper; wash the precipitate two or three times with water until it becomes free from chloride. All the air-dried content with the filter paper is transferred to the already weighed silica crucible and placed into an electric muffle furnace (Temperature about 1000⁰C) for ignition and weighed as calcium oxide. Finally, the percentage of calcium is calculated in terms of the drug taken.

RESULTS:

All values are reported here as the mean of three or more observations.

Table No. 2: Organoleptic Properties

S. No.	Properties	Results
		KBM (L)	Commercial samples
		KBM (L)	KBM (C-1)	KBM (C-2)
1.	Color	White	Off - White	White
2.	Odor	Odourless	Odourless	Odourless
3.	Taste	Lime like	Lime like	Lime like
4.	Touch	Smooth	Smooth	Smooth
5.	Appearance	Lustreless	Lustreless	Lustreless

Table No. 3: Preliminary Test of KBM

S. No.	Tests	Results
		KBM (L)	Commercial samples
		KBM (L)	KBM (C-1)	KBM (C-2)
1.	Floating	Positive	Positive	Positive
2.	Fitness	Positive	Positive	Positive
3.	Wallstick	Positive	Positive	Positive
4.	Finger	Positive	Positive	Positive

Table No. 4: Physicochemical parameters analysis

S. NO.	Parameters analyzed	Results (Mean ± SEM) %
		KBM (L)	Commercial samples
		KBM (L)	KBM (C-1)	KBM (C-2)
1	Bulk density (gm/ml)	0.51± 0.00	0.50± 0.00	0.52± 0.00
2.	Tapped Density (gm/ml)	0.92± 0.02	0.90± 0.00	0.97± 0.00
3.	Hausner’s ratio (HR)	1.80 ± 0.02	1.8 0± 0.02	1.86 ± 0.03
4.	Compressibility index	44.50 ± 0.08	44.44± 0.05	46.39 ± 0.04
5.	(a)pH 1% Aqueous solution (b)pH 10% Aqueous solution	11.98 ± 0.00 12.10 ± 0.00	12.10± 0.00 12.10± 0.00	11.99 ± 0.00 12.00 ± 0.00
6.	Loss on Drying at 105⁰ C	0.010± 0.00	0.015± 0.00	0.019 ± 0.00
7.	Total ash (%)	98.80 ± 0.06	98.50± 0.02	95.50± 0.05
8.	Acid Insoluble ash (%)	0.25 ± 0.02	0.95± 0.00	1.50 ± 0.00
9.	Water soluble ash (%)	3.50 ± 0.02	4.50± 0.03	6.50 ± 0.00
10.	Water insoluble ash	95.50 ±0.01	93.5 ± 0.02	92.5 ± 0.02
11.	Extractive values (%) a) Distilled water b) 1N Hydrochloric acid	5.80 ± 0.17 98.60 ± 0.25	4.50 ± 0.00 99.05± 0.00	6.50 ± 0.02 94.50 ± 0.0
12.	Estimations (%) (a) Calcium (Ca) (b) Chloride (Cl)	35.70 + 0.12 0.27± 0.02	31.20 ± 0.00 0.43± 0.03	39.50 ± 0.20 0.35 ± 0.10
13.	Sedimentation time (min.)	5.7 ± 0.00	5.2 ± 0.00	6.00 ± 0.00

Table No. 5: Qualitative Elemental Analysis

S. No.	Elements	Results
		KBM (L)	Commercial samples
		KBM (L)	KBM (C-1)	KBM (C-2)
1.	Sodium	Present	Present	Present
2.	Potassium	Present	Present	Present
3.	Calcium	Present	Present	Present
4.	Magnesium	Present	Present	Present
5.	Chloride	Present	Present	Present
6.	Carbonate	Present	Present	Present

DISCUSSION:

Morphologically the color of KBM-L was found to be white, smooth to the touch in texture, odorless, and lime-like in taste (Table No 2) while two other classical samples KBM(C-1) and KBM(C-2) have the same characteristics except a minor color difference in KBM(C-1) which was off-white in color instead of white. Some more classical tests like floating, fineness, and thumb finger test (fig. 4) were found to be positive (Table 3) in both commercial and lab-made samples which indicates that prepared kushta is perfect as per classical parameters. As far as scientific parameter is concerned as given in Table no. 4, the values of density (bulk and tapped density) were reported in gm/ml, these largely depend upon particle shape which is inversely proportional to particle size i.e. High value indicates a very small particle size¹⁹. The mean values of bulk and tapped density were 0.51 gm/ml and 0.92gm/ml respectively in the lab-made sample and this value was very close to KBM(C-1) but slightly differing from 0.52 and 0.97gm/ml in the case of KBM(C-2). The values of the Hausner ratio and Carr’s compressibility index were found to be 1.80 and 44.50 for the lab-made sample while 1.80 and 44.44 for KBM (C-1) and 1.80 and 46.39 for KBM (C-2). A value of more than 1.2 of the Hausner ratio indicates poor flow properties²⁰. Sedimentation time of KBM(L), KBM(C-1) and KBM(C-2) was 5.7, 5.2 and 6.00 minutes respectively. The pH values of 1% and 10% aqueous solutions for KBM(L) / KBM(C-1)/ KBM (C-2) were 11.98 and 12.10/ 12.10 and 12.12/11.99 and 12.00 respectively (Table no. 3) which are very similar. It is mentioned that most of the kushtajat are alkaline²¹. It may also be concluded that the finished product was a content of metallic oxide i.e. Calcium oxide (CaO) as the fact that pH value of water solutions of metal oxide is basic ²². The mean percentage of loss of weight on drying was negligible as prepared kushta showed very little weight loss on drying. It could be assumed that the finished product was devoid of water and organic matter. The mean percentage of total ash, acid-insoluble ash, water-soluble ash, and water-insoluble ash was 98.80, 0.25 3.50 and 95.5% respectively while samples KBM(C-1)/ KBM(C-2) have these values 98.5, 0.95, 4.50 and 93.5 / 95.50, 1.50, 6.50 and 92.5% respectively. High ash value shows the presence of very high inorganic content. In case the extractive value of kushta is performed to extract outorganic matter if present²³. The mean percentages value of water and 0.1N hydrochloric acid soluble extractives was21.80and 98.6in KBM (L), in KBM(C-1) is 24.50 and 99.05 while it was 26.50 and94.50 % in KBM (C-2) respectively. These extractive values help in determining adulteration and are an index of the purity of the drugs. Less extractive value again confirms that kushta was prepared properly and the finished product was free from organic material. The analytical results of KBM (L) /KBM(C-1)/ KBM (C-2) showed that the mean % of Calcium and chloride was 35.70 and0.27 /31.20 and0.43 and 39.50 and 0.35 respectively. It indicates that the majority of the kushta was in oxide form. Qualitative tests of inorganic elements showed the presence of Calcium, Magnesium, Sodium, Potassium, Carbonate, and Chloride as mentioned in Table No. 5.

CONCLUSION:

Despite the great advancement of technologies scientific work on this type of preparation like Kushta has been lacking which may be due to a lack of proper communication among traditional healers, clinicians, and scientists due to the difference in concept, etc. The comparative data of physical findings obtained in the present study is a trial between market samples and the product produced in the laboratory by using the Standard Operative Procedures (SOP). However, testing on classical parameters and most of the other physical parameters for both lab-made and commercial samples were found to be positive and values are in range. No difference was found regarding the presence of inorganic elements during the qualitative analysis but in the case of the quantitative estimations, considerable variation has been found. These studies indicate going ahead towards the comparative clinical evaluation of the samples by way of doing the pharmacological studies which are needed to be done on KBM to convince the conventional society regarding the safety and high efficacy of Unani compound formulation.

With all that, it may be concluded that the study provides valuable information regarding the methods of preparation and physicochemical parameters on classical as well as modern techniques which may be taken as standard parameters for future reference. In other words, it can be stated that the physicochemical methods adopted herein to evolve the standards, will serve as a tool to control the quality of the drug in the manufacturing sector and in dispensing pharmacy.

REFERENCES:

1. Prakash VB. Importance of metallic micronutrients in Ayurveda conference on Health and Environment. 1988. 1-8

2. SaeedM, Hamdard Pharmacopeia Karachi, Hamdard Academy 1969: 222-234

3. Rasheed A, Marri A, Naik MM. Standardization of Bhasma; Importance and Prospects, Journal of Pharmacy Research. 2011; 4(6): 1931-33

4. Ferraz Edwardo Gamelas, Jose A.F. Coronado Joao, MonteiroCarlos; Roche Fernando (2018-09-03) “Eggshell waste to produce building time” Calcium oxide reactivity, Industrial Environmental and economic implications. Materials and Structures. 51(5) der: 10.1617/511527-018-1243

5. Mazlam N. A Sapari J. M, Sambasevam K.A. Synthesis and fabrication of polyaniline eggshell composite in ammonia detection. Journal of Metals Materials and Minerals. 30(2): 50-57.

6. Magyar Cheryl. Ruralsprout.Com/egg shell-uses/ Published on June 27, 2019

7. National Formulary of Unani Medicine, Part I, CCRUM, Ministry of AYUSH, Govt of India; Pp69.

8. Kabiruddin, M. Bayaz-e-Kabir Part I, Sheikh Jamia, JamiaTibbiya Delhi; P 113

9. Hafeez, A.Sanatul Taklees, CCRUM, Ministry of AYUSH, Govt of India; P 234-235

10. Kabiruddin, M. Bayaz Hakim-e-Kabir, Part III, Hyderabad, Hikmat Book Depot; Deccan 24-25, 119

11. Farzana A, Halder S, Shuma ML and Rouf AS. In vitro Release Kinetics study of Indomethacin 2HR. Metrix Tablet from Methoeel K4M CR and Methoeel K15M CR. Jour of Biomed and Pharm Res. 2012; 13: 60:65

12. Nuka R Potu AR and Nandan R. Formulation Development and In-vitro evaluation of Ramipirill Micropellets. World Journal of Pharma Res. 2012; 2(1): 76-87

13. Ramanjaneyeulu M, Kumar MS, Bairam R, and Babu PM. Formulation and evaluation of oral disintegrating Release Dosage form containing meclizine HCl. International Journal of Pharma. 2012; 2(4): 717-726

14. Anonymous. Physicochemical Standards of Unani Formulations, CCRUM, New Delhi. 2006: 142-145

15. Anonymous. The Unani Pharmacopeia of India, Ministry of AYUSH, Govt of India, New Delhi. Vol 2, 2007:116

16. Anonymous. The Ayurvedic Pharmacopeia of India, Ministry of AYUSH, Govt of India, New Delhi. Vol2, 2008:160

17. Anonymous. Quality Control Methods for Herbal Materials; Geneva, 2011:31

18. Caven, R. M; Quantitative Chemical Analysis and Inorganic preparations, 2nd edition, Blackie and Son Limited, London Glasgow, 1962, pp. 44-45

19. Lechman L, Liberman HA, and Kanug JL. The Theory and Practice of Industrial Pharmacy, Verghese Publishing House, Mumbai, 1991: 316

20. Sultan EM. Aultons Pharmaceutics, Churchill Livingstone, London. 2009: 176-178

21. Qasmi IA. Kitabul Taklees, Aligarh Muslim University, Aligarh, YNM: 36.

22. Pant KM, Gill JS, Jain SC, Joshi R, Agarkar SC, and Srivastava VP. Science and Technology Class 8th Publication Department NCERT New Delhi

23. Rasheed A, Marin A, and Naik MM. Standardisation of Bhasma; Importance and Prospects. Journal of Pharma Res. 2011; 4(6): 1931-1933

Received on 28.02.2024 Modified on 23.03.2024

Asian J. Pharm. Tech. 2024; 14(2):113-118.

DOI: 10.52711/2231-5713.2024.00020