Syeda Nishat Fathima*1, S. Vasudeva Murthy2
1Department of Pharmacy, Shri Jagdishprasad Jhabarmal Tibrewala University, Rajasthan, India
2Department of Pharmacology, Jayamukhi College of Pharmacy, Warangal, Telangana, India
Address for Corresponding Author:
Syeda Nishat Fathima,
Department of Pharmacy, Shri Jagdishprasad Jhabarmal Tibrewala University,
Rajasthan, India
Abstract
Objective: The purpose of the present study was to explore the Pharmacognostic parameters for standardization of Rosa Damascena Petals. Material and Methods: The flowers of Rosa Damascena were authenticated and shade dried. Petals were separated and powder characteristics, behavior and fluorescence analysis, physiochemical assessment, and micrometric investigation were carried out. Dried Rosa Damascena Petals powder was then extracted with aqueous, alcoholic, chloroform, petroleum ether, and ethyl acetate solvents and their extractive values were calculated. The phytochemical screening of Rosa Damascena petals was carried on all the different extracts. Results: The characteristic macroscopic and microscopic features of petals include the presence of wavy epidermal cells, spirally thickened xylem vessels, lignified fibers, oil globules, prismatic, acicular clusters and conglomerate crystals of calcium oxalate and Cystoliths crystals of calcium carbonate. Powder showed characteristic fluorescent property when treated with different reagents. Physicochemical exploration showed values for moisture content, moisture sorption capacity, ash values and extractive values which are within the limits of World Health Organisation standards for the crude drug from medicinal plants. Micromeritic analysis of petal powder reveals good flowability. Aqueous and ethanolic extractive values were found to be higher when compared to that of extractive values of chloroform, petroleum ether, and ethyl acetate. Preliminary Phytochemical exploration indicated the presence of carbohydrates, glycosides, alkaloids, flavonoids, amino acids, and triterpenoids. Conclusions: The current research would be useful in order to supplement the information regarding pharmacognostical characteristics, physiochemical evaluation, micrometric analysis and phytochemical exploration in Ayurvedic system of medicine for its identification.
Keywords: Rosa damascena, physicochemical evaluation, fluorescence analysis, phytochemical screening
Introduction
Nowadays there is a renewed interest in drugs of natural origin merely because they are deliberated as green medicine and green medicine and are thought to be safe. In addition to safety, tolerability, effectiveness, expenditures especially in long-term treatment, serum drug monitoring etc. are other limitations with synthetic drugs. The advantage of drugs from natural sources is their easy availability, economic and less or no side effects but the disadvantage is that they are the victims of adulteration. The more effective the natural drug more is its demand and the chances of non-availability upsurges. To meet with the growing demand, the natural drugs are easily adulterated with low-grade material. Pharmacognostic studies ensure that the drug identity lays down standardization parameters which will help and prevents the adulterations. Such studies will help in authentication of the plants and ensures reproducible quality of herbal products which will lead to safety and efficacy of natural products (Sumitra, 2014).
Rosa damascena, known as Damask rose, a perennial bushy shrub, is the most famous ornamental plant and the holy ancient herb with novel applications belonging to the Rosaceae family. Its vernacular names include Gulab in Hindi, Gulaabi poovvu in Telugu and Mahakumari or Satapatri in Sanskrit. Chemical composition revealed the presence of Citronellol, geraniol, nerol, phenyl ethyl alcohol; nonadecane, nonadecene, eicosane, heneicosane, tricosane, a-guaiene, geranyl acetate, and eugenol have been reported (Mohaddese, 2016). The most beneficial effects of R. damascena in ancient medicine are including treatment of abdominal and chest pain, strengthening of the heart, treatment of menstrual bleeding and digestive problems, and reduction of inflammation, especially of the neck, cough remedy for children, gentle laxative as well as Rose oil heals depression, grief, nervous stress, tension, allergies, headaches, and migraine. Some of the proved pharmacological properties include anti-HIV, antibacterial, antioxidant, antitussive, hypnotic, antidiabetic, and relaxant effect on tracheal (Mohammad et al., 2011). Apart from this gel of Rosa damascena has been proved for UV Protective activity (Patil et al., 2011).
The objective of the present study is to evaluate various pharmacognostical parameters such as macroscopic, microscopy, physicochemical, fluorescence and phytochemical studies of the Rosa damascena petals.
Materials and methods
Collection and authentication of plant material
The fresh flowers of the deciduous shrub of Rosa damascena were collected in bulk from the local area of Warangal, Telangana, India. The flowers were authenticated by Dr. P. Veera Reddy, Professor, Government Ayurvedic College, Warangal, Telangana. The petals were separated from the sepals and shadow dried.
Macromorphological description of Rosa damascena petals
The Rosa damascena petals were subjected to macroscopic studies which comprised of organoleptic characteristics such as color, odor, taste, texture, shape, and size of the drug (Anita et al., 2016).
Microscopic characteristics of Rosa damascena petals
Microscopic sections of Rosa damascena petals were cut by freehand cross-sectioning, temporary mounts of the sections of the petals were stained with saffranine, mounted with glycerine water and observed under a microscope (Harinarayan et al., 2007).
Powder characteristics of Rosa damascena petals
On the clean glass slide, fine powder of Rosa damascena petals was stained with Phloroglucinol-HCl and Sodium hypochlorite solution mounted with glycerin water. The slide was then placed and observed under the magnifying lenses of 10x, 40x and 100x magnification of the microscope (Zhongzhen, 2010).
The behavior of Rosa damascena petals Powder with chemical reagents
The behavior of Rosa damascena petals with different chemical reagents was performed to detect the occurrence of phytoconstituents along with color changes under ordinary daylight by a standard method (Pratt and Chase, 1949)
Fluorescence analysis of Rosa damascena petals
The Rosa damascena petals powder was placed on a clean microscopic slide and 2 to 3 drops of freshly prepared reagents (acids such as 1 N HCl and 50% H2SO4; and alkaline solutions such as aqueous sodium hydroxide, alcoholic sodium hydroxide; and other solvents such as nitric acid, picric acid, acetic acid, ferric chloride, and nitric acid with ammonia) were added and mixed by gentle tilting the slide. They were then exposed to fluorescence analysis in the ultraviolet (UV)-light (254 nm and 365 nm) (Dharamveer et al., 2013; Kokashi et al., 1958)
Physico-Chemical evaluation of Rosa damascena petals
Physiochemical parameters such as moisture content, pH, ash constants (Saroja, 2009) and soluble extractive values (Sundar and Justin, 2016) on Rosa damascena petals were performed according to the official method prescribed and the WHO guidelines on quality control methods for medical plants Material (Prathapa et al., 2015).
Micromeritic evaluation of Rosa damascena petals
The micromeritic characteristics of Rosa damascena petals powder like Bulk density, Tapped Density, Angle of repose, Hausner’s ratio and Carr's index was determined according to the official standard procedures to study the flowability of the drug (Martin, 1994).
Preparation of extracts of Rosa damascena petals
The dried powder material (100 g) of the Rosa damascena petals was powdered and passed through sieve no.16. These powder petals were macerated with water, ethanol, chloroform, ethyl acetate, and petroleum ether for 7 days with occasional shaking in between. The extracts were filtered through muslin cloth, then the filtrate was evaporated under reduced pressure, vacuum dried and stored. The preliminary Phytochemical screening of the different extracts was then carried out (Mohanty et al., 2011).
Preliminary phytochemical screening of Rosa damascena petals
The preliminary phytochemical screening was carried out on the different extracts of Rosa damascena petals for the detection of various phytochemicals such as Carbohydrates, Alkaloids, Glycosides, Saponins, flavonoids, proteins, amino acids, tannins, fixed oil, fats, steroids and terpenoids (Khandelwal, 2008; Mohammad et al., 2013).
Results and discussion
Macrmorphologcal description of Rosa damascena petals
The Rosa damascena flowers are light to moderate pink to light red with relatively small flowers grow in groups. The results of macromorphology were shown in table 1. The Fresh Petals, Dried, and powdered Rosa damascena flowers were illustrated in figure 1 whereas microscopy of the flower is shown in figure 2.
Figure 1. Fresh petals, dried, and powdered Rosa damascena flowers
Table 1. Macrmorphological description of Rosa damascena flowers
|
S. No |
Characters |
Observation |
|
Organoleptic Characters |
||
|
1. |
Colour |
Magenta on base and light yellow near to apex |
|
2. |
Odor |
Aromatic Distinct |
|
3. |
Taste |
Distinct tongue sensitizing aromatic taste with pleasant mild sweetness |
|
Quantitative Macromorphology of Fresh petals |
||
|
4. |
Width |
0.9-3.8cm |
|
5. |
Length |
1.8-4.2cm |
|
Extra Features |
||
|
6. |
Shape |
Heart/ Pear Shape |
|
7. |
Texture |
Soft and smooth |
Microscopic characteristics of Rosa damascena petals
Figure 2. Microscopy of Rosa damascena petals
Some of the microscopical powder characteristic features that are seen in Rosa damascena petals are wavy epidermal cells, spirally thickened xylem vessels, lignified fibers, oil globules, prismatic, acicular clusters and conglomerate crystals of calcium oxalate and Cystoliths crystals of calcium carbonate.
Figure 3. Powder Characteristics of Rosa damascena petals: (a) Spirally thickened xylem vessels, (b) Phloem fibers, (c) Lignified fibers, (d) Lignified fibers, (e) Cystoliths crystals of calcium carbonate, (f) Acicular clusters of calcium oxalate, (g) Conglomerate crystals of calcium oxalate, (h) Oil gland, (i) Spongy Parenchymatous cells, (j) Xylem Parenchymatous cells with crystal, (k) Pollen grains, (l) Hairy Trichome
Behavior and Fluorescence analysis of Rosa damascena petals
Fluorescence is the significant phenomenon exhibited by many chemical constituents existing in the natural products. Some display fluorescence in the visible range in daylight. The ultraviolet light produces fluorescence in many plant materials which do not markedly fluoresce in daylight. If the plant material is not fluorescent in nature it can show fluorescence after treated with different reagents, henceforth it can be used as an important parameter for qualitative assessment in the pharmacognostical evaluation. The results of Behavior and fluorescent analysis of Rosa damascena petals of powder and different extracts showed characteristic coloration in treatment with various chemical reagents.
Table 2. Behavior and Fluorescence analysis of Powder of Rosa damascena petals
|
S. No |
Treatment |
Day Light |
UV Light (254nm) |
UV Light (365nm) |
|
|
Powder as such |
Pale violet red |
Olive drab |
Black |
|
|
Powder + Conc. HCl |
Scarlet |
Olive |
Bister |
|
|
Powder + Conc.HNO3 |
School bus yellow |
Lawn green |
Black |
|
|
Powder + Conc. H2SO4 |
Dark Rust |
Olive drab |
Dark brown |
|
|
Powder + Conc. NaOH |
Blaze orange |
Green Yellow |
Black |
|
|
Powder + Ethanol |
Pale magenta |
Olive drab |
Black |
|
|
Powder + Glacial acetic acid |
Carmine |
Medium Violet red |
Crimson |
|
|
Powder +Dil. NaOH |
Amber |
Lawn green |
Black |
|
|
Powder + Picric acid |
Orange |
Lawn green |
Bister |
|
|
Powder +FeCl3 |
Black |
Dark Olive green |
Black |
|
|
Powder + NaHCO3 |
Dark olive green |
Olive drab |
Black |
|
|
Powder + 5% I2 |
Burnt Orange |
Olive drab |
Black |
Table 3. Behavior and Fluorescence analysis of extracts of Rosa damascena petals
|
S. No |
Treatment |
Day Light |
UV Light (254nm) |
UV Light (365nm) |
|
|
Water |
Cardinal |
Dark Corel |
Black |
|
|
Ethanol |
Carmine |
Bister |
Black |
|
|
Pet Ether |
Pear |
Lawn Green |
Olive drab |
|
|
Chloroform |
Yellow |
Bright Green |
Olive |
|
|
Ethyl acetate |
Tangerine |
Green |
Black |
The consistency and extractive values of different extracts of Rosa damascena petals
An extractive value signifies the number of constituents present in the given amount of plant material extracted with different solvents. It provides an indication of the extent of polar, medium polar and non-polar constituents present in the crude drug. In the present study water-soluble extractive value and alcohol soluble extractive values were found to be more when compared to petroleum ether, chloroform, and ether soluble extractive values, which denotes that Rosa damascena petals contain more quantity of polar compounds.
Table 4. The consistency and extractive values of different extracts of Rosa damascena petals
|
S. No. |
Treatment |
Consistency |
Extractive values (%) |
|
|
Water |
Sticky |
13.16 |
|
|
Ethanol |
Sticky |
17.24 |
|
|
Pet Ether |
Sticky |
4.68 |
|
|
Chloroform |
Sticky |
2.12 |
|
|
Ethyl acetate |
Powder |
6.96 |
Physicochemical parameters of Rosa damascena petals
Loss drying is used to estimate the amount of volatile matter including water that is present in the plant material. Percent Loss on drying was found to be 14.32%. The moisture content of a drug should be reduced in order to avoid decomposition of crude drugs, either due to chemical change or microbial contamination. The percentage of moisture content ranging from 10 - 20% indicates an ideal range for bacteria as well as for fungal growth. The extent of polysaccharide that is present in certain drugs is denoted by swelling index. It is one of the characteristics for identification of botanical drugs if swelling index changes it indicates that the powder has been adulterated or not properly stored. For the present drug, the swelling factor in water after 24 hours was found to be 4.9 ml. The ability of a drug to take up water is determined by moisture sorption capacity. Greater the moisture sorption capacity higher will be chances for bacterial or fungal contamination. In the present study moisture sorption capacity of Rosa damascena petals were found to be 0.72/gm in 24 hours. After three days of keeping the sample in the desiccator fungal growth was observed due to excess moistness.
Ash value is a benchmark to judge the identity or purity of crude drugs. The total ash residue remaining after incineration which usually represents the inorganic salts naturally occurring in the plant material and adhering to it, but it may also include inorganic matter added for the purpose of adulteration. Ash value is a useful tool for detecting low-grade products or exhausted products or excess of sandy or any earthy substance with the drug. Acid-insoluble ash denotes the presence of only earthy matter i.e., sand or silica in the drug whereas Water soluble ash detects the drug exhausted with water, if admixed with the exhausted material will show a much greater reduction in water-soluble ash than total ash. So, it’s an important indicator when exhausted material is substituted for the genuine drug. In the present study Total ash, Acid insoluble ash and Water-soluble ash were found to be 6.34, 1.51 and 2.48 % w/w respectively.
Table 5. Physicochemical parameters of Rosa damascena petals
|
S. No |
Constants |
Yield (N=3) |
|
|
Foreign matter |
0 |
|
|
Moisture content( Loss on drying) % |
14.32 % |
|
|
pH |
6.56 |
|
|
Swelling Index ml |
4.9 |
|
|
Moisture Sorption Capacity /g |
0.72 |
|
|
Total ash (% w/w) |
6.34 |
|
|
Acid insoluble ash (% w/w) |
1.51 |
|
|
Water soluble ash (% w/w) |
2.48 |
Micrometric parameters of Rosa damascena petals
Bulk density, Tapped Density, Angle of repose, Hausner’s ratio and Carr's index were determined as a part of the micrometric analysis. The Carr’s compressibility index and Hausner’s ratio gives the insight value of the difference in the bulk and tapped densities. While the Carr’s index shows strength and the ability of a drug to reduce in volume, the Hausner’s ratio reveals inter particulate friction in between particles. As the values, these indices decrease the flow property of the powdered drug increases. The angle of repose is a traditional characterization method for determining the flow property of powder. The result showed that the powder has good flowability as the angle of repose of powder was found to be 31.21 0.
Table 6. Micrometric parameters of Rosa damascena petals
|
S. No |
Constants |
Yield |
|
|
Bulk density |
0.202 g/ml |
|
|
Tapped Density |
0.259 g/ml |
|
|
Angle of repose |
31.21 0 |
|
|
Hausner’s ratio |
0.779 |
|
|
Carr's index |
22.22 % |
Preliminary phytochemical screening of different extracts of Rosa damascena petals
Phytochemical screening of Rosa damascena petals showed that maximum phytoconstituents are present in ethanolic and aqueous extract extracts.
Table 7. Preliminary phytochemical screening of different extracts of Rosa damascena petals
|
S. No |
Plant constituent |
Aqueous Extract |
Alcohol |
Pet Ether |
Chloroform |
Ethyl acetate |
|
|
Alkaloids |
Positive |
Positive |
Negative |
Negative |
Positive |
|
|
Glycosides |
Positive |
Positive |
Negative |
Positive |
Positive |
|
|
Carbohydrates |
Positive |
Positive |
Negative |
Negative |
Negative |
|
|
Flavonoids |
Positive |
Positive |
Negative |
Positive |
Positive |
|
|
Tannins |
Positive |
Positive |
Negative |
Negative |
Positive |
|
|
Proteins |
Positive |
Positive |
Negative |
Negative |
Positive |
|
|
Amino acids |
Positive |
Positive |
Negative |
Negative |
Positive |
|
|
Fixed oils |
Negative |
Negative |
Positive |
Positive |
Negative |
|
|
Sterols |
Positive |
Positive |
Positive |
Positive |
Positive |
|
|
Starch |
Positive |
Negative |
Negative |
Negative |
Negative |
|
|
Cardiac glycosides |
Positive |
Positive |
Negative |
Negative |
Positive |
Conclusion
In conclusion, the pharmacognostical data on the Rosa damascena petals can assist as a relevant source of information and contribute towards the standards for its identification and authentication.
Acknowledgment
Authors are sincerely thankful to Management of Jayamukhi College of Pharmacy, Warangal, India for providing necessary facilities and encouragement to carry out the research work successfully.
Conflicts of interest: Not declared.
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