PHARMACOGNOSTIC, PHYTOCHEMICAL AND PHYSICO-CHEMICAL EVALUATION OF LEAVES OF TRAGIA INVOLUCRATA LINN.,
HTML Full TextPHARMACOGNOSTIC, PHYTOCHEMICAL AND PHYSICO-CHEMICAL EVALUATION OF LEAVES OF TRAGIA INVOLUCRATA LINN.,
M. Thenmozhi *, M. Gokul and K. Muthu Krishnan
Department of Pharmacology, Captain Srinivasa Murthy Central Ayurveda Research Institute, Chennai, Tamil Nadu, India.
ABSTRACT: Background: Tragia involucrata Linn., leaves are traditionally used for the treatment of liver disorders, eczema, viral infections, headache, respiratory infections, etc. There is lack of documentation of safety data and non-availability of quality control standards for the ayurvedic traditional medicines. Aim & Objective: The aim of the study was designed to determine the pharmacognostic, phytochemical and physico-chemical standardization of leaves of Tragia involucrata Linn. Materials & Methods: The Pharmacognostical parameters such as organoleptic evaluation (macroscopy) and microscopy (T.S of Leaf) were done to identify the morphological and anatomical structures. For powder microscopy, coarse powder is treated with routine reagents to identify the diagnostic features of the plant. Preliminary phytochemical screening, physicochemical parameters was determined as per the prescribed guidelines and fluorescent behaviour of coarse leaf powder was also determined. These parameters are useful tools to differentiate the powdered drug material from the adulterated ones. Results: The macroscopical characters such as colour, shape, size, odour and surface characteristics were observed. Transverse section of leaf and powder microscopy revealed the presence of spongy parenchyma, paracytic stomata, glandular trichomes, etc. Phytochemical screening showed the presence of tannins, volatile oils, flavonoids, saponins, triterpenes and glycosides. Physicochemical parameters such as moisture content, ash value, extractive value, etc and fluorescent behaviour of leaf powder were determined. Conclusion: The data collected from the present study will definitely contribute to develop quality standards of Tragia involucrata and serve as a baseline for planning future studies.
Keywords: Physico-chemical, Phytochemical, Powder microscopy, Organoleptic, Tragia involucrata
INTRODUCTION: The significance and recognition of traditional ayurvedic medicines, herbal remedies, and formulations are on the rise globally.
There is a growing demand for herbal medicines in both developed and developing nations due to their diverse biological properties, increased safety profile, and affordability for all socioeconomic groups.
However, the lack of proper standardization methods and quality control has led to the production of finished products without adherence to good manufacturing practices 1. In order to change the perspective of inferior quality of drug manufacturing practices and create products that benefit the public, it is essential to establish quality standards for herbal formulations, natural products, and other related items.
The purpose of this study is to establish quality standards for Tragia involucrata leaves and to verify the standardization parameters in accordance with WHO guidelines. The collection of plant material, along with factors such as time, temperature, drying method, moisture content, geographical and seasonal variations, are crucial for ensuring good manufacturing practices in the production of ayurvedic formulations and herbal medicines 2. Each step in the process of preparing raw drugs requires careful attention. Tragia involucrata, a member of the Euphorbiaceae family, has traditionally been utilized for treating liver disorders, eczema, viral infections, headaches, respiratory infections, and more. This study aims to assess the pharmacognostic characteristics, conduct preliminary phytochemical screening, and analyze the physico-chemical properties of Tragia involucrata Linn.
MATERIALS & METHODS:
Plant Material Identification and Authentication: Tragia involucrata leaves were collected from Kasampatti, Natham in Dindigul district and authenticated (Authentication no: R0138) by Dr. Stephen, Professor in the Department of Botany at American College in Madurai-20. The herbarium of this specimen has been preserved in the department for further reference.
Pharmacognostical Evaluation: The characteristics of leaves, including colour, smell and taste, are examined through organoleptic testing following specific guidelines. Fresh leaves were chosen for microscopic analysis by cutting them in cross-section. In powder microscopy, a coarse powder is created and treated with standard chemicals to identify unique plant features 3.
Physio-chemical Parameters: The powder undergoes analysis to determine its physiochemical characteristics including the presence of foreign organic matter, moisture content, ash content, and extractable components using solvents of varying polarity. The testing method followed the standard prescribed guidelines 4.
Fluorescence Analysis: Behavioural characters of Tragia involucrata Linn. (Leaf-Crude Powder) was determined with water and different reagents such as Conc. HCl, Conc. H2SO4, Conc. HNO3, CH3COOH, aqueous NaOH and aqueous FeCl34.
Preparation of the Extract: Approximately 250 g of the dried leaves of Tragia involcucrata was coarsely powdered and soaked in the menstruum which consists of 70:30 Ethanol: Water. The solvent/menstruum was selected based on the order of increasing polarity. The solvent was poured on to the drug material until it is completely soaked and kept for 3 days with occasional shaking to achieve complete extraction. Then the supernatant was decanted carefully and evaporated to dryness on a water bath. The concentrated extract residue was found to be 5g. The percentage yield was found to be 3.99%.
Qualitative Analysis: The hydroalcoholic extract of Tragia involucrata Linn (Leaf) was analyzed through a series of chemical tests to determine the presence of various compounds 5.
RESULTS & DISCUSSION:
Pharmacognostic Evaluation: The Organoleptic characteristics of was showed in the Table 1 and the macroscopy of leaves-dorsal and ventral view of Tragia involucrata was showed in Fig. 1. The cross-section of Tragia involucrata leaves revealed distinctive traits, including circular petioles covered by a cuticle and sparse elongated, glandular capitate trichomes. These trichomes featured elongated biseriate stalks and club-shaped secretory bodies. The epidermal layer consisted of small, square-shaped cells.
The parenchymatous ground tissue was thin walled, with circular cells that were less compact at the edges and larger, angular cells that were more tightly packed towards the center. Additionally, numerous rosette crystals were observed throughout. The vascular system appeared multi-stranded, with a ring of distinct collateral vascular strands containing a high concentration of xylem and a lower concentration of phloem elements, as shown in Fig. 2. These assessment methods assist in distinguishing between pure samples and adulterated ones based on their unique characteristics 6.
TABLE 1: MACROSCOPICAL STUDIES OF TRAGIA INVOLUCRATA LINN (LEAF)
S. no. | Parameters | Observation |
1 | Colour/surface | Dorsal-Darkgreen; Ventral-Lightgreen |
2 | Odour | Characteristic dour |
3 | Taste | Characteristic taste |
4 | LeafType | Simple |
5 | Shape | Ovateorelliptic |
6 | Arrangement | Alternate |
7 | Apex | Acuminate |
8 | Base | Acuteorrounded |
9 | Stipules | Present |
10 | Margin | Serrate |
11 | Venation | Pinnate |
12 | Surface | Stinging hairs |
13 | Length | 6cmto 10cm |
14 | Width | 3.5cmto 5cm |
15 | Petiole length | 2.5cm long |
16 | Flower | Yellow |
FIG. 1: MACROSCOPY OF TRAGIA INVOLUCRATA LINN LEAF
FIG. 2: T.S OF LEAF OF TRAGIA INVOLUCRATA. E–Epidermis; Chl–Chlorenchyma; Col–Collenchyma; GT–Glandular Trichome; Ph–Phloem; Pi–Pith; Rcr–Rosettecrystal; T–Trichome;Ve–Vessel.
FIG. 3: T. S OF LAMINA THROUGH MIDRIB. LE–Lowerepidermis, Me–Mesophyllcells, Pal–Palisadecells, Ph–Phloem, Rcr–Rosettecrystal, SP–Spongy Parenchyma, VB–Vascular Bundle, UE–Upperepidermis, Xy–Xylem.
Powder Microscopy: The powder microscopy of the plant material is buff in colour, unicellular trichome, glandular trichome, paracytic stomata and showed crystals of calcium oxalate and Xylem vessels as shown in Fig. 4.
FIG. 4: POWDER MICROSCOPY
Physicochemical Evaluation: The various physicochemical parameters of leaves of Tragia involucrate i.e., Loss on drying, Total solids, petroleum ether extractive, Ethyl acetate extractive, chloroform extractive, ethanol extractive, aqueous extractive, Total ash value water soluble ash and acid insoluble ash was showed in Table 2.
Total ash is the measure of the total amount of material left after burning and includes ash derived from the part of the plant and adherent material. The total ash value for Tragia invoucrata samples was 0.223333±0.017639%w/w, which was in the limit (not more than 15%). The acid-insoluble ash is the residue obtained after boiling the total ash with dilute hydrochloric acid and burning the remaining insoluble matter 7. The process measures the amount of silica present, especially in the form of sand and siliceous earth. The value of acid-insoluble ash was 0.60 ± 0.034749 %w/w, which is lesser than 5% and thus passes the test. Water-soluble extractives are indicative of water-soluble active constituents of crude drugs, such as tannins, sugars, plant acids, mucilage, glycosides etc. The water-soluble extractive value is 9.003333 ± 0.702405 % w/w. Alcohol is an ideal solvent for extraction of various chemicals like tannins, resins etc and the value was found to be 3.966667 ± 0.833818 % w/w. The moisture content of a drug should be minimized in order to prevent decomposition of crude drugs, either due to chemical change or microbial contamination 8. The percentage of moisture content ranging from 10 - 20% shows an ideal range for minimum bacteria as well as for fungal growth. Ash value is a criterion to judge the identity or purity of crude drugs. The Loss on drying value was found to be 1.98 ± 0.863178 % w/w. The powder did not possess any foreign matter, bitter principle. The powder also exhibits petroleum ether extractive value (1.656667±0.331689%w/w) Ethylacetate extractive value (2.323333±0.689376 % w/w) Chloroform extractive value (1.873333 ± 0.218359 % w/w).
TABLE 2: DETERMINATION OF PHYSICO-CHEMICALPARAMETERS OF TRAGIA INVOLUCRATA (LEAF)
S. no. | Physico–Chemical constant | Reports |
1 | Foreign Matter | NIL |
2 | Bitterness Value | NIL |
3 | Loss on Drying | 1.98± 0.863178 %w/w |
4 | Total solids | 98.02± 0.704782 %w/w |
5 | Petroleum ether extractive | 1.656667± 0.331689 % w/w |
6 | Ethyl acetate extractive | 2.323333± 0.689376 %w/w |
7 | Chloroform extractive | 1.873333± 0.218359 % w/w |
8 | Ethanol extractive | 3.966667± 0.833818 % w/w |
9 | Aqueous extractive | 9.003333± 0.702405 % w/w |
10 | Total ash | 0.223333± 0.017639 % w/w |
11 | Water soluble ash | 3.46± 0.110591 %w/w |
12 | Acid in soluble ash | 0.60± 0.034749 %w/w |
Preliminary Phytochemical Screening: A preliminary analysis was conducted on the hydroalcoholic extract to identify phytochemical compounds. The findings are detailed in Table 3. Qualitative screening of the drug was performed to detect alkaloids, flavonoids, saponins, tannins, and proteins.
TABLE 3: PRELIMINARY PHYTOCHEMICAL SCREENING OF HYDRO-ALCOHOLIC EXTRACT OF TRAGIA INVOLUCRATE (LEAF)
S. no. | Name of the test | Haeti |
1 | Test for carbohydrate | |
Fehling’s test | Negative | |
Molisch test | Positive | |
Benedict’s Test | Positive | |
2 | Test for proteins and free amino acid | |
Biuret test | Positive | |
Millon’s Test | Negative | |
Ninhydrin Test | Negative | |
Sulphur Containing Amino acid | Negative | |
3 | Test for Flavonoides | |
Ferricchloride test | Positive | |
Shinoda test | Positive | |
Alkaline reagent test | Positive | |
Zinc-Hydrochloric acid reduction test | Negative | |
4 | Test for alkaloids | |
Mayer’s test | Positive | |
Wagner’s test | Positive | |
Dragendorff’s test | Positive | |
Hager’s test | Negative | |
5 | Test for saponins | |
Foamtest | Positive | |
6 | Test for sterols | |
Salkowaski test | Positive | |
Libbermann Burchard test | Positive | |
7 | Test for triterpenoids | |
Salkowaski test | Positive | |
8 | Test for Glycosides | |
Keller-Killiani test | Positive | |
Baljet’s test | Positive | |
Legal test | Positive | |
Raymond’s test | Negative | |
Bromine water test | Negative | |
9 | Test for Tannins | |
Ferric-chloride test | Positive | |
Gelatin test | Positive | |
10 | Test for fats | |
Solubility test | Negative | |
Filter paper test | Negative | |
11 | Test for volatile oils | |
Odor test | Positive | |
12 | Test for Mucilage | Positive |
13 | Test for Coumarin | Negative |
14 | Test for Resins | Negative |
15 | Test for Gum | Negative |
16 | Test for Quinone | Negative |
17 | Test for Anthocyanin | Negative |
Hydroalcoholic extract of Tragia involucrate (Leaf) was subjected to qualitative chemical analysis. The various chemical tests were performed on this extract for the identification of phytochemicals, secondary metabolites.
Fluorescence Analysis: The alterations in the behavior of powdered drug with distinctive chemical reagents were analyzed under both UV and visible light, as detailed in Table 4.
TABLE 4: BEHAVIOURAL CHARACTERS OF THE TRAGIA INVOLUCRATA
Crude powder of Tragia involucrata (Leaf) | Visible light | UV Light (254nm) | UV Light (365nm) |
Powder+Water | Light green | Dark black | Dark green |
Powder+Conc.Hcl | Dark green | Black | Green |
Powder +Con.H2SO4 | Black | Dark green | Black |
Powder+HNO3 | Black | Green | Dark green |
Powder+CH3COOH | Light black | Brown | Darkgreen |
Powder+Con.HCL+Water | Light green | Light black | Dark green |
Powder+Con.H2SO4+Water | Dark black | Light brown | Light black |
Powder+Con.HNO3+ Water | Light black | Dark brown | Light green |
Powder+CH3COOH+ Water | Light black | Light brown | Dark green |
Powder+aqueousNaOH | Light green | Black | Dark green |
Powder+aqueous FeCl3 | Dark black | Black | Dark green |
TABLE 5: QUANTITATIVE MICROSCOPY OF TRAGIA INVOLUCRATA LEAVES
S. no. | Parameters | Upper epidermis | Lower epidermis |
1 | Epidermal Number | 60/mm2 -65/mm2-70/mm2 | 70/mm2-72/mm2 – 75mm2 |
2 | Stomatal Number | 132/mm2 -133/mm2-135/mm2 | 152/mm2-155/mm2 –158/mm2 |
3 | Stomata Index | 65/mm2-66/mm2-68/ mm2 | 67/mm2-67/mm2-68/mm2 |
4 | Palisade Ratio | 10/mm2-12/mm2 – 14/mm2 | 9/mm2-10.5/mm2-12/mm2 |
5. | Vein Islets | 8/mm2-9/mm2 -10/mm2 | |
6. | Vein termination | 8/mm2-10/mm2-12mm2 |
FIG. 5: QUANTITATIVE MICROSCOPY. E–Epidermis; St– Stomata, VI–Veinislet, VT–Veintermination
ACKNOWLEDGEMENT: I am grateful for the support, direction, and motivation provided to me by my colleagues and staff members.
CONFLICT OF INTEREST: The Authors have no competing interests to declare.
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How to cite this article:
Thenmozhi M, Gokul M and Krishnan KM: Pharmacognostic, phytochemical and physico-chemical evaluation of leaves of Tragia involucrata Linn. Int J Pharm Sci & Res 2024; 15(8): 2338-45. doi: 10.13040/IJPSR.0975-8232.15(8).2338-45.
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IJPSR
M. Thenmozhi *, M. Gokul and K. Muthu Krishnan
Department of Pharmacology, Captain Srinivasa Murthy Central Ayurveda Research Institute, Chennai, Tamil Nadu, India.
thenprabhu2014@gmail.com
17 February 2024
04 April 2024
19 April 2024
10.13040/IJPSR.0975-8232.15(8).2338-45
01 August 2024