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 Table of Contents  
Year : 2015  |  Volume : 7  |  Issue : 4  |  Page : 250-253  

Development of standard operating procedure and standardization of Habb-e-Banafsha Qawi-A Unani polyherbal formulation

1 Department of Biotechnology, Faculty of Science, Centre for Transgenic Plant Development, Jamia Hamdard, New Delhi, Uttar Pradesh, India
2 Department of Saidla, Aligarh Unani and Ayurvedic Medical College and A C N Hospital, Aligarh, Uttar Pradesh, India
3 Department of Pharmacognosy, Faculty of Pharmacy, Jamia Hamdard, New Delhi, Uttar Pradesh, India
4 Department of Ilmul-Advia, Faculty of Medicine, Jamia Hamdard, New Delhi, Uttar Pradesh, India

Date of Submission10-Apr-2014
Date of Decision01-Jan-2015
Date of Acceptance15-Feb-2015
Date of Web Publication23-Oct-2015

Correspondence Address:
M Z Abdin
Department of Biotechnology, Faculty of Science, Centre for Transgenic Plant Development, Jamia Hamdard, New Delhi, Uttar Pradesh
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/0975-7406.168019

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Introduction: Herbals drugs became a boon for mankind since ancient times and still are used worldwide for the treatment of various human ailments. The safety of alternative medicinal preparations has been questioned due to reports of unwanted side effects. To maintain the quality and efficacy of these drugs, it is essential to standardize them in order to avoid the use of substandard or adulterated medicines in the market. Unani system of medicine mainly focuses on the treatment by natural drugs. Habb-e-Banafsha Qawi is useful in a cough, catarrah, and coryza. Materials and Methods: Physiochemical constants of Habb-e-Banafsha Qawi were determined through organoleptic characters, macro- and micro-scopic characters, ash value, solubility, pH values. Chromatographic fingerprints were developed using thin layer chromatography method. Aflatoxin (AF) contamination, heavy metal, and pesticide residues were also evaluated by standard methods.Objectives: In the present study, an attempt has been made to develop standard operating procedure and physiochemical parameters to monitor the quality of a Unani poly-herbal formulation, Habb-e-Banafsha Qawi. Results: The tablets tasted sweetish bitter with a pleasant odor, water soluble and acidic in nature. Rfvalues were almost same in all the extracts. No AF, heavy metal, and pesticide residues were observed. Conclusions: It may be concluded that the values and chromatographic fingerprints obtained can be used for quality evaluation and to set new pharmacopoeial standards for the preparation of Habb-e-Banafsha Qawi.

Keywords: Habb-e-Banafsha Qawi, herbals drug, standard operating procedure, Unani poly-herbal formulation

How to cite this article:
Ali A, Sumbul S, Ahmad MM, Ahmad S, Kabir H, Abdin M Z. Development of standard operating procedure and standardization of Habb-e-Banafsha Qawi-A Unani polyherbal formulation. J Pharm Bioall Sci 2015;7:250-3

How to cite this URL:
Ali A, Sumbul S, Ahmad MM, Ahmad S, Kabir H, Abdin M Z. Development of standard operating procedure and standardization of Habb-e-Banafsha Qawi-A Unani polyherbal formulation. J Pharm Bioall Sci [serial online] 2015 [cited 2021 Jan 20];7:250-3. Available from:

According to an estimate of World Health Organization (WHO), around 85–90% of the world's population consumes traditional herbal medicines, directly or indirectly.[1] Despite of the rising popularity of herbal drugs, has also given rise to many reports describing adverse health effects due to variable quality, efficacy, and contents of herbal products as they are a class of pharmaceutical products and should meet the requirements of quality, safety and efficacy.[2],[3],[4] Adulterations with toxic compounds, contamination with pathogenic microbes or presence of natural toxins like aflatoxins (AFs) have been reported in herbal products and plants.[5],[6],[7],[8],[9],[10],[11],[12] In this regard, WHO had developed several guidelines for carrying out standardization procedures of raw herbal products to standardize certain herbal products.

Habb-e-Banafsha Qawi is composed of Banafsha (viola odorata Linn.), Turbud (Marsdenia tenacissima), Rubb-us-Soos (extract of Glycyrrhiza glabra Linn.) and Kateera (Cochlospermum religiosum). It is an important Unani compound drug useful in a cough, catarrah and coryza. In our present study, attempts have been made for standardization of Habb-e-Banafsha Qawi following the scientific parameters including organoleptic characters, macro- and micro-scopic characters, thin layer chromatography (TLC), ash value, solubility, pH values, AF contamination, heavy metal, and pesticide residues.

   Materials and Methods Top

Plant materials

Ingredients, after collection from the market (M/S Shamsi Dawakhana, Ballimaran, Delhi), were authenticated by Dr. H.B. Singh, Department of Herbarium and Museum, NISCAIR, New Delhi.

Preparation of tablets

The crude drugs were grounded into powdered depending upon the plant. The powder was then, passed through the mesh sieve number 100 (British Standard Sieve, BSS). The tablets were prepared on the instructions of National Formulary of Unani Medicine.

Organoleptic characters

These parameters were determined through observations made by a panel of 10 healthy individuals (five male and five female) in the age group of 22–30 years.

Macro and micro-scopic evaluation

The shape, texture, and consistency of the tablet were measured through touch and visual observations while weight and dimensions were measured using microbalance and vernier calipers, respectively.

One tablet of Habb-e-Banafsha Qawi was crushed, powdered and stained with safranin. A mount was prepared to examine under compound microscope and structures were then drawn.

Thin layer chromatography

Extracts were prepared by extracting 2.0 g of powdered tablet with 40 ml each solvent (ethanol, petroleum ether [60–80°C] and chloroform), respectively for 6 h at 37°C with 60 rpm. A volume of 2 µl of each sample solution was applied to the absorbent layer of silica plate by sample applicator (Linomat 5, CAMAG, Switzerland). The TLC plate was then, developed with 50 ml of mobile phase consisting of toluene:ethyl acetate (8:2 v/v) for 90 min. The developed plate was derivatized with anisaldehyde reagent (Abs. ethanol:glacial acetic acid:conc. H2 SO4:anisaldehyde in the ratio of 85:10:5:0.5, v/v/v/v). The plate was photographed and scanned using Wincats 3.00 software (CAMAG, Switzerland), at the visible range at 550 nm.

Determination of ash values

Total, acid insoluble and water soluble ash

The powdered drug 5.0 g was incinerated in a silica crucible in a muffle furnace at a temperature not exceeding 450°C. The samples were ignited until the color changed to white, which was then cooled and weighed to get the total ash content. The process was repeated until a constant weight was obtained, and the percentage of ash was calculated.

The total ash obtained was boiled with 25 ml dilute HCl (10%) for 5 min on a water bath. The insoluble ash was collected on Whatman Filter Paper and washed with hot water and ignited at a temperature not exceeding 450°C and the percentage of acid insoluble ash was calculated.

The same amount of total ash was dissolved in MilliQ water (25 ml) and boiled for 5 min on a water bath. The remaining process was repeated as performed with acid insoluble ash and percentage of water soluble ash was calculated.

Solubility studies

Solubility in alcohol, water, and petroleum ether

The powdered tablet 5.0 g was dissolved in 100 ml of absolute ethanol (99%), water (MilliQ) and petroleum ether (60–80°C), respectively, in a closed flask and shaken at regular interval for 24 h and then allowed to stand for 18 h at room temperature. It was then filtered through Whatman Filter Paper. The insoluble sample along with filter paper was kept in an oven at 105°C for drying until a constant weight was obtained. The percentage of alcohol-soluble, water-soluble and petroleum ether soluble matter was calculated by following formula:

pH values

The powdered drug (1.0 g) was dissolved in MilliQ water to make 1.0 and 10% w/v solutions and pH was measured on the digital pH meter (Orion, USA).

Aflatoxin contamination

AFs were extracted from a ground sample with 70% methanol. The extracted sample and enzyme-conjugated AF were mixed and added to the antibody-coated microwell. Further steps were performed according to the manufacturer's instructions. The plate was measured optically with an absorbance filter (OD450) and a differential filter of 630 nm and were compared with the standards.

Determination of pesticide residue

The organochlorine pesticides were determined by extracting each sample in Soxhlet apparatus and the extract were run under gas chromatograph (GC) instrument. Identification and quantification were accomplished using known amount of external standard procured from Sigma-Aldrich. However, the determination of organophosphorus pesticides was performed by GC-mass spectrometry.

Analysis of heavy metals

Heavy metals were estimated by following the protocols of Rai et al[13] Analysis was done by using atomic absorption spectrophotometer (AAS; Perkin Elmer 5000). Qualitative and quantitative analysis of mercury and arsenic was performed following the Espinoza et al. on X-ray fluorescence spectrometer and AAS, respectively.[14]

   Results and Discussion Top

The standardization and quality control of herbal materials by use of modern science and technology is vital. The organoleptic evaluation was done for identification of sensory characteristics like color, odor, taste, size. Extractive values are an approximate measure of the amount of a certain constituent or a group of certain constituents; the drug contains.[15] Organoleptic characters, macroscopic characters, ash values, solubility, and pH values were determined, and their observations are depicted in [Table 1]. The tablets appeared in the form of solids with muddy color. The odor was found to be pleasant and tasted sweetish bitter [Table 1]. Microscopic examination of powdered drug specimen, as depicted in [Figure 1], revealed that it was mainly comprised of fragments of fibers, single or in group (Banafsha, Turbud). In a surface view, epidermal cells of Banafsha were observed which were made up of small rectangular to squarish cells. Fragments of vessels either single or in a group with simple pits were seen (Turbud). Total, acid insoluble and water soluble ash values were found to be 1.70 ± 0.60, 1.03 ± 0.31 and 0.83 ± 0.16%, respectively. Like most of the other tablets, Habb-e-Banafsha Qawi tablets have the highest solubility with water (9.16 ± 0.04%) followed by petroleum ether and ethanol. The pH values of both (1.0 and 10%) solutions of tablets were found to be acidic in nature. Chromatographic fingerprinting techniques are most significant methods which can be used for the routine herbal drug analysis and for quality assurance. By analytical techniques such as TLC, HPLC, and HPTLC, the presence of compounds can also be assessed and quantified to a certain limit.[16],[17],[18] The details of TLC fingerprint of Habb-e-Banafsha Qawi are shown in [Figure 2] and [Table 2]. Contamination with AFs and pesticides can alter the quality, safety, and efficacy of herbal drugs. No AFs (B1, G1) were found. However, traces of B2 and G2 were found in the tablets which were less than the permissible limit. Contamination by heavy metals and pesticides in herbal remedies can pose clinically relevant dangers for the health of the user and should therefore, be in limit.[19] However, during the experiments, no heavy metals and pesticides were observed.
Table 1: Physiochemical constants of Habb-e-Banafsha Qawi

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Figure 1: Microscopic structure of Habb-e-Banafsha Qawi: Fibers (a), vessels (b) and epidermis (c)

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Figure 2: Thin layer chromatography fingerprint of Habb-e-Banafsha Qawi. L1–L2: Ethanolic extract, L–L4: Petroleum ether extract, L5 and L6: Chloroform extract

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Table 2: Rf values of Habb-e-Banafsha Qawi

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   Conclusion Top

Development of standard operating procedures and standardization is of great significance in assuring the quality, authenticity, as well as purity and thereby, the efficacy of the drug. Standardization of drugs is essential to provide good quality drugs of high efficacy and potency. Such kind of standardization studies are needed in promoting the safe use of genuine drugs, thus contributing to human health. The methods used for standardization help in identification and collection of genuine drugs and discard of the adulterated, spurious drugs. Besides the economic loss, the use of the sub-standard drug can cause serious adverse effects in the body. Thus, it can be concluded that the parameters studied in the present work may be used for quality evaluation and results obtained could be used to lay down a new set of pharmacopoeial standards for the preparation of Habb-e-Banafsha Qawi to obtain optimal efficacy of the medicine.


AA and SS are thankful to CCRUM for the providing fellowships.

Financial support and sponsorship


Conflicts of interest

There are no conflicts of interest.

   References Top

WHO Report. WHO/EDM/TRM/2002. Geneva: World Health Organization; 2002. p. 19.  Back to cited text no. 1
Calixto JB. Efficacy, safety, quality control, marketing and regulatory guidelines for herbal medicines (phytotherapeutic agents). Braz J Med Biol Res 2000;33:179-89.  Back to cited text no. 2
Capasso F. The medicinal plants in our time. Boll Chim Farm 1986;125:322-7.  Back to cited text no. 3
Taylor DA. Botanical supplements: Weeding out the health risks. Environ Health Perspect 2004;112:A750-3.  Back to cited text no. 4
Boullata JI, Nace AM. Safety issues with herbal medicine. Pharmacotherapy 2000;20:257-69.  Back to cited text no. 5
Elvin-Lewis M. Should we be concerned about herbal remedies. J Ethnopharmacol 2001;75:141-64.  Back to cited text no. 6
Martins HM, Martins ML, Dias MI, Bernardo F. Evaluation of microbiological quality of medicinal plants used in natural infusions. Int J Food Microbiol 2001;68:149-53.  Back to cited text no. 7
Kneifel W, Czech E, Kopp B. Microbial contamination of medicinal plants – A review. Planta Med 2002;68:5-15.  Back to cited text no. 8
Reif K, Metzger W. Determination of aflatoxins in medicinal herbs and plant extracts. J Chromatogr 1995;692:131-6.  Back to cited text no. 9
Halt M. Moulds and mycotoxins in herb tea and medicinal plants. Eur J Epidemiol 1998;14:269-74.  Back to cited text no. 10
Efuntoye MO. Mycotoxins of fungal strains from stored herbal plants and mycotoxin contents of Nigerian crude herbal drugs. Mycopathologia 1999;147:43-8.  Back to cited text no. 11
Elshafie AE, Al-Rashdi TA, Al-Bahry SN, Bakheit CS. Fungi and aflatoxins associated with spices in the Sultanate of Oman. Mycopathologia 2002;155:155-60.  Back to cited text no. 12
Rai V, Agarwal M, Khatoon S, Rawat AK, Mehrotra S. Estimation of Co and Mn in some medicinal plants. Bull Environ Contam Toxicol 2001;66:427-32.  Back to cited text no. 13
Espinoza EO, Mann MJ, Bleasdell B, DeKorte S, Cox M. Toxic metals in selected traditional Chinese medicinals. J Forensic Sci 1996;41:453-6.  Back to cited text no. 14
Afaq SH, Tajuddin, Siddiqui MM. Standardization of Herbal Drugs. Aligarh: AMU Press; 1994. p. 1-24.  Back to cited text no. 15
Wallis TE. Textbook of Pharmacognosy. 5th ed. New Delhi, India.: CBS Publishers and Distributors Pvt. Ltd; 2001. p. 563.  Back to cited text no. 16
Huie CW. A review of modern sample-preparation techniques for the extraction and analysis of medicinal plants. Anal Bioanal Chem 2002;373:23-30.  Back to cited text no. 17
Sapna SK, Ravi TK. Approaches towards development and promotion of herbal drugs. Pharmacogn Rev 2007;1:180-4.  Back to cited text no. 18
World Health Organization. Basic Tests for Drugs, Pharmaceutical Substances, Medicinal Plant Materials and Dosage Forms. Geneva: World Health Organization; 1998.  Back to cited text no. 19


  [Figure 1], [Figure 2]

  [Table 1], [Table 2]


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