Studies Leading to Phyto and Physico-chemical Evaluation of an important Polypharmaceutical preparation (Syrup) "Sharbat Toot Siyah"- A Drug of Choice

 

Akhlaq Mustafa¹*, Umar Hussain², Anas Iqbal Alvi³, Ghazala Javed⁴, Asim Ali Khan⁵

¹Research Officer (Chemistry), Drug Standardization Research Unit,

Central Council for Research in Unani Medicine, Janakpuri, New Delhi.

²Senior Research Fellows (SRF), Drug Standardization Research Unit,

Central Council for Research in Unani Medicine, Janakpuri, New Delhi.

³Research Assistant (Chemistry), Drug Standardization Research Unit,

Central Council for Research in Unani Medicine, New Delhi.

⁴Research Officer Incharge (Unani),

Central Council for Research in Unani, Medicine, 61-65, Institutional Area, Janakpuri, New Delhi.

⁵Director General, Central Council for Research in Unani Medicine, 61-65, Institutional Area, Janakpuri, New Delhi.

 

ABSTRACT:

Unani system of medicines classifies its formulation into several classes e. g. Huboob, Sufoof, Qurs, Sufoofs, Majoon, Arq, Roughan, Kushta etc. Among the various dosage forms of Unani system of medicine Sharbat Toot Siyah is one of the renowned formulations whichis widely used for the number of ailments e.g. Pharyngeal pain and sore throat. The drug Sharbat Toot Siyah was in-house manufactured in three batches of different volumes of 500, 600 and 700ml following the SOP guidelines and underwent for the evaluation of secondary metabolite constituents Profile with Physico-chemical Standardization of this important poly-herbal formulation. The present work contributes to the development of standardization parameters of herbal drugs used in Indian system of medicine like morphological properties, viscosity, pH values, ash values, extractive values, alcohol soluble matter, specific gravity at 25ºC, refractive index, sugar quantity e. g. total sugar, reducing sugar and non-reducing sugar, qualitative and quantitative inorganic elements, phyto-constituents and thin layer chromatography. In preliminary phytochemical investigation, analysis showed the presence of organic constituent’s amino acids, tannins, phyto-sterols, flavanoids and glycosides in the extract while in the elemental analysis, calcium, magnesium, phosphorous etc. were found to be present. The study also includes quality control parameters e. g. aflatoxins, microbial load, pesticide residue and detection of heavy metals which are helpful to ensure the purity, safety and efficacy of herbal formulation. The results so obtained for the various Physico-chemical tests may be taken as standard values for future reference.

 

 

 

 

 

INTRODUCTION:

The origin of Sharbat is buried in antiquity. Traditionally its invention has been attributed to Pythagoras, the ancient Greek philosopher and mathematician. The incorporation of various ingredients into a Sharbat prolongs the life of medicinal ingredients. Also, due to the palatability of Sharbat, the patient is not reluctant to drink it. However, it must be born in mind that the Sharbat described here serves as a vehicle for appropriate medicines prescribed for the ailments¹. Sharbat is a popular drink in West and South Asia prepared from fruits or flower petals. It is sweet in taste. Popular Ashriba is made of one or more of the following: rose water, sandalwood, bael, gurhal, lemon orange mango pineapple and falsa. In the 12^th century, Ismail Jurjani mentioned in his book Zakhirah Khwarazm shahi different types of Ashriba in Iran, including gur, anar, sikanjbin, and so forth². It is a sweet aqueous liquid made by dissolving sugar, misri, honey, or jaggery in water. But according to Qarabadeen, Al-Qarabadeen, Bayaz-e-kabeer, Qarabadeen Majeedi, Qarabadeen Jadeedi, Sharbat is a sweet viscous liquid made by preparing the decoction or infusion of the ingredient drugs or by taking out the juice of the fruit which is then mixed with sugar and boiled to a required consistency (Qiwam) that is of one tar.The equivalent of Sharbat in a modern system is syrup. Syrups are concentrated, viscous, aqueous solutions of sugar substitute with or without flavors and medical substances^3-7. These are used to treat coughs and sore throats, relieve tickling and irritation of the throat, loosens phlegm, facilitate expectoration, and heal and soothe the throat and lungs. When purified water alone is used in making the solution of sucrose, the preparation is known as syrup or simple syrup if the sucrose concentration⁸ is 85%. Among the long list of numerous Sharbats; Sharbat Toot Siyah is also an important Unani formulation and has been given in Bayaz-e- Kabeer, Qarabadeen-e-Jadeedi and National Formulary of Unani Medicine, part-1. The action of the drug is anti-inflammatory, demulcent and therapeutically uses are acute hoarseness, tonsillitis, laryngitis bronchitis and catarrh⁹.

 

Formula composition of this preparation contain only one ingredient of plant origin, is Toot Siyah known as mulberry in English and belongs to the genus Morus of the family Moraceae. It is an economically important plant being used for sericulture, as it is the sole food plant for the domesticated silkworm, Bombyxmori. The genus Morus, is widely distributed in Asia, Europe, North America, South America, and Africa, and is cultivated extensively in the eastern, central, and southern Asia for silk production. Mulberry is found from temperate and sub-tropical regions of the northern hemisphere, as well as in the tropics of the southern hemisphere, because it can grow in a wide variety of climatic, topographical, and soil conditions. They spread throughout all regions from the tropics to the sub-arctic and from sea level to altitudes as high as 4000m. Mulberries are grown at considerably higher altitudes in the Himalaya-Hindu Kush region, while in Pakistan they are widely cultivated in northern regions^10-13. The drug has an antioxident, Neuroprotective, Antitumor, Immunomodulative activities¹⁴.

 

Keeping in view the importance and popularity of this effective Unani medicine among the masses an attempt has been made here to develop the physico-chemical standards of Sharbat Toot Siyah by doing the study on three different batch sizes of different volumes 500 ml, 600 ml and 700 ml for a consistent chemical profile or simply a quality assurance program for production and manufacturing of herbal formulations.

 

Formulation composition:

As per the formula given in the following table the drug was prepared

 

Table no. 1: Ingredients of Sharbat Toot Syiah

 

MATERIALS AND METHODS:

The crude drug was procured from the local market, Delhi and authenticated by Pharmacognosy section of Drug Standardization Research unit, New Delhi. Three batches of the poly-herbal formulation were prepared as per the formula mentioned in National Formulary of Unani Medicine Part-I, 2006.

 

Method of preparation:

For preparation of the drug, take the ingredient of pharmacopoeia quality. Clean the ingredients and make fine powder. Mix ingredient numbered 2 in ingredient number 1. Boil the mixture till the required consistency is reached. Filter the mixture and store in a sterile container and make it air-tight to protect from light and moisture. Three batches of different volumes; 500ml of batch-1, 600 ml of batch-2 and 700 ml of batch-3 were prepared as per the composition given above in table no. 1.

 

RESULTS:

Organoleptic characters:

Sharbat Toot Siyah is a liquid compound preparation with dark brown color sweet in taste and characteristic odor is given as in table no. 2.

 

Table No. 2: Organoleptic Characters

 

 

Physico and phyto-chemical evaluation of Sharbat Toot Siyah:

Physico-chemical parameters e.g. total ash, acid Insoluble ash, alcohol and hexane soluble extractives, pH values of 1 and 10% aqueous solution along with the specific gravity, viscosity, refractive index, sugar estimations were carried out as per the method described in WHO guidelines¹⁵.

Table no. 3: Quantitative physico / phytochemical constants of Sharbat Toot Siyah

Values are expressed as mean % ± SD except pH values are expressed as mean ±SD

 

The study of quantitative estimation of organic constituents present in the drug was conducted as per described method¹⁶. For the chromatographic estimations, thin layer chromatography of ethanol extract was carried outand data of Rf values are reported by following the method on percolated Silica gel 60 F264 TLC plates^17-19.

 

Table no. 4: Qualitative analysis of phyto-constituents in alcoholic Extract of Sharbat Toot Siyah

Indications: (-ive) Absent and (+ive) Presence of constituent

 

 

Quality Control Parameters of Sharbat TootSiyah:

In order to ensure the quality of drug, the modern techniques and standard methods were adopted. The parameters such as microbial load and heavy metals were carried out as per WHO guidelines. Aflatoxins and Pesticide residues carried out by standard method²⁰. The heavy metals viz. lead, cadmium, mercury and arsenic were found to be negative while aflatoxins B1, B2, G1, and G2 were not detected and pesticide residues DDT and Endosulfan were found below the limit of quantification. Other parameters like estimation of microbial load viz. Escherichia coli and Total Fungal count (TFC) were not detected while Total Bacterial load and Salmonella spp were found to be within the permissible limit as stated by W H O.

Table no. 5: Quantitative estimations of Heavy Metals

 

Table no. 6: Analysis of Microbial Load

 

Table no. 7: Estimations of Aflatoxin

 

Table No. 8: Analysis of Pesticide Residues

Indications: LOQ (Limit of quantification)

 

Chromatographic estimation Sharbat TootSiyah:

Thin Layer Chromatography:

Extract of ethanol of the drug was spotted on a pre-coated silica gel 60 F254 TLC (Merck Germany) as an absorbent and developed the plate using Toluene: Ethyl acetate (8: 2) as a mobile phase. After developing, plate was dried and studied under UV 254nm, 366nm as well as spraying with 2% Vaniline - Sulphuric acid followed by heating at 105⁰C for 10 minutes in an oven²¹and reported the Rf values of spots observed as given in the following table no 9.

 

Table No. 9: Chromatographic Evaluation

 

Qualitative elemental analysis:

Sodium (+), Potassium (+), Calcium (+),

Phosphorous (+), Iron (+), Zinc (+) and

Magnesium (+),

 

DISCUSSION:

As per the formula composition, the polyherbal preparation contains only two ingredients. Out of these one belongs to the plant origin drug (Fruits part of M. indica L.) and other one is crystalline sugar or sucrose in solid form. Organoleptic characteristics of the compound formulation indicate that compound preparation is in liquid form with dark brown color sweet in taste and characteristic odor (Table no. 2). Phytochemical screening of the chemical constituents present in the drug revealed that it contains glycosides, steroids, phenolic, tannins, proteins, flavonoids, amino acids, carbohydrate and alkaloids while the saponins was found to be negative. These phyto-constituents so obtained during the analysis has been shown in (Table no. 4) for finished product. In quantitative estimation for physico-chemical constants & phytochemical constituents, three experiments were conducted for each sample and their % mean values of total phenolic, tannins, and alkaloids are mentioned in (Table no. 3). Estimation of ash values are an important criterion for judging the impurities along with identity and quality of the drugs because adulteration of herbal drugs with unwanted materials like earthy matters resulting in higher ash percentage. The values of the total ash indicate that the amount of mineral matter present in the drug. Water and alcohol soluble extractives indicate the presence of inorganic content while alcoholic soluble extractives indicate the extraction of polar constituents. The pH values of all three samples were found to be acidic. In quality control parameters the bacterial load & total fungal count of microbial studies, aflatoxin, heavy metals and pesticide residues were either within the permissible limits or found to be absent as shown in Table nos. 5, 6, 7 and 8 respectively. At the same time bacteria Enterobacteriaceae, Salmonella spp, Escherichia coli, Staphylococcus aureus, were found to be absent which shows that the drug is not the carrier of these organisms. TLC profile of ethanol extracts can be seen in table              no. 9.

 

CONCLUSION:

The presented result of phyto-chemical screening, physico-chemical, and other parameters of all three batches of different volumes of polyherbal formulation Sharbat Toot Siyah shows that there is no variation in the batch to batch consistency of the drug. Besides, the study of quality control parameters revealed the absence of microbial load, heavy metals, aflatoxins and pesticide residue contamination which shows that the drug possesses non-toxic effect and genuine raw drug material were added in the formulation and role of these analyzed parameters are very helpful to minimize the chances of adulteration of this useful drug. Therefore it can be concluded that evaluated data presented here will be much useful and serve not only to assess the quality of drug but also will reflect the purity of ingredients used in the drug.

 

ACKNOWLEDGEMENT:

The authors are grateful to the Director General, Central Council for Research in Unani Medicine for providing necessary facilities. The authors are also thankful to DSRI Ghaziabad for providing the necessary assistance.

 

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Received on 10.09.2022       Modified on 02.11.2022

Accepted on 18.12.2022   ©AandV Publications All Right Reserved