Background: Many medicinal plants, including Spinacia oleracea, Withania somnifera, Emblica officinalis, and Caesalpinia bonducella, are helpful in treating anaemia, a malady that kept them in inadvertent isolation. Aim and objective: The goal of this study is to establish the macro- and micro-morphological standards as well as the pharmacognostic, phytochemical, and physicochemical standards for clear distinctions between the chosen plants. Materials and Methods: Macro- and micro-morphological features, quantitative microscopy, soluble extractives, pH, ash values, and phytochemical profiles of leaves of S. oleracea and C. bonducella, roots of W. somnifera, and fruit of E. officinalis were determined using standard methods. Results: The leaves of S. oleracea and C. bonducella leaves were smooth, succulent, ovate to triangular or elliptic-oblong, and green in hue. The roots of W. somnifera are stout, long, woody, tuberous, fleshy, and of a whitish-brown color, and the E. officinalis fruit had a greenish yellow color and was smooth, spherical, or globular in shape. Additional differences in stomata arrangement, epidermal cell, histological features of the leaf's midrib, W. somnifera root, and E. officinalis fruit, as well as the physicochemical and phytochemical profiles of S. oleracea and C. bonducella leaves, W. somnifera root, and E. officinalis fruit, provide helpful information for clearly differentiating adulteration. Conclusion: These are important for making sure those manufacturers, regulators, and researchers of herbal products get the right plants.
Keywords: Antianemic, Caesalpinia bonducella, Emblica officinalis, microscopy, Spinacia oleracea, Withania somnifera
|How to cite this URL:|
Thumar MM, Pethani TM, Patel NV. Pharmacognostic evaluation on four commonly used antianemic plants: Spinach, amla, ashwagandha, and kakachiya. J Integr Health Sci [Epub ahead of print] [cited 2023 Mar 22]. Available from: https://www.jihs.in/preprintarticle.asp?id=365628
| Introduction|| |
Anemia is a common nutritional deficiency disorder and global public health problem that affects both developing and developed countries with major consequences for human health and their social and economic development. Anemia is a general term applied to the condition in which there is insufficient or defective formation of hemoglobin and defective maturation and formation of red blood cells. The normal hemoglobin level is 13 g/dl for men and 12 g/dl for women, which has been reduced in anemia. Acquired causes of anemic states are poor diet, low socioeconomic status, high exposure to blood parasites, and a helminthic infection. The selection of herbal drugs over allopathic drugs is increasing worldwide for the prevention and treatment of various health challenges, such as diabetes, hypertension, anemia, and cancer. To meet the growing demand, the natural drug is easily forged with inferior-quality materials. Misuse of medicinal plants or natural products begins with misidentification. To tackle this problem, pharmacognostic study and the standardization of medicinal plants play an important role. Therefore, this study focuses on macro- and micromorphological standards, phytochemical, physicochemical, and pharmacognostic studies of some medicinally important plants that are traditionally used for the treatment of anemia.
The chosen plants are Spinacia oleracea, Withania somnifera, Emblica officinalis, and Caesalpinia bonducella, among the many medicinal herbs that play a beneficial role in an anemic state. S. oleracea has been used for many different illnesses in the past, such as leprosy, asthma, biliousness, diabetes, maturation, antipyretic, cooling, emollient, laxative, diuretic, anthelmintic, and inflammation of the lungs and bowels. W. somnifera may also be used to treat coughs, bronchitis, senile dementia, rheumatism, asthma, swollen glands, arthritis, anxiety, stress, and constipation. It may also be used as a narcotic and an aphrodisiac. E. officinalis is used to treat a variety of conditions, including hemorrhoids, leprosy, anemia, cough, asthma, dysentery, diarrhea, helminthic infections, hyperacidity, dyspepsia, colitis, gastritis, osteoporosis, menorrhagia, and diabetes. Numerous traditional medicinal applications of C. bonducella include tonic, anthelmintic, febrifuge, laxative, leprosy, malaria, colic, tumors, inflammation, and liver issues.
Caesalpinianone and 6-O-methylcaesalpinianone, on the other hand, have been isolated from C. bonducella. S. oleracea has yielded spinacetin, patuletin, and jaceidin. The chemically diverse plants are rich in various types of flavonoids such as quercetin, kaempferol, myricetin, luteolin, and apigenin. E. officinalis has been linked to phenolic substances such as coumaric acid, gallic acid, ellagic acid, pyrogallol, and chlorogenic acid. Numerous clinically beneficial alkaloids have been reported in the past, including nicotine, withanine, withananine, withasomnine, somniferinine and somnine from W. somnifera, as well as phyllantine and phyllantidine from E. officinalis., Various secondary metabolites, including phytosterols, terpenoids, and minerals, have also been reported.
| Materials and Methods|| |
Plant collection and processing
The leaf of S. oleracea and C. bonducella, the root of W. somnifera, and the fruit of E. officinalis were collected in February and March around Rajkot, Gujarat. They were taxonomically identified and authenticated by Dr. V. S. Thaker, Department of Bioscience, Saurashtra University, Rajkot, Gujarat, India (SU/Bio/1063). The fresh plant material was collected thoroughly cleaned and air-dried. It was then homogenized to a fine powder and stored in air-tight bottles for further studies.
Micro- and macromorphological features
The assessment of S. oleracea and C. bonducella leaves, W. somnifera root, and E. officinalis fruit was carried out for their shape, size, surface properties, texture, color, consistency, aroma, and taste, among other organoleptic and macroscopic qualities. Additionally, the leaf's morphology and arrangement were examined.
By using conventional methods, free-hand sections were taken for the pharmacognostic examination. Using phloroglucinol in hydrochloric acid to mount the specimen, transverse sections of selected parts of S. oleracea, W. somnifera, E. officinalis, and C. bonducella were examined under a microscope (×10 and ×40) to look for collenchyma, lignified tissues, and other distinguishing characteristics. Chloral hydrate solution was used as a clearing agent.
The fruit of E. officinalis, the root of W. somnifera, and the leaves of S. oleracea and C. bonducella were all finely powdered before being examined under a microscope with phloroglucinol in concentrated hydrochloric acid. Photomicrographs were taken of cellular characteristics.,
According to the WHO guideline, physicochemical examination was carried out using coarse powder of certain parts of S. oleracea, W. somnifera, E. officinalis, and C. bonducella. Total ash value, loss on drying, water-soluble ash, acid-insoluble ash, extractive values, and pH were determined. The amount of water and alcohol-soluble components was calculated using the aqueous and alcoholic extractive values. Ash value, extractive value, and pH were performed in triplicate.
Using the cold maceration process, the dried, powdered plant material was extracted with methanol and water (6:4). The extracts were filtered and concentrated using a rotary evaporator. Using the methods described, a preliminary phytochemical screening of the extracts was done to see if they had specific phytoconstituents such as alkaloids, glycosides, carbohydrates, phytosterols, fixed oils, saponins, phenolic compounds, tannins and flavonoids, proteins, and amino acids.,
| Results|| |
Plant description and macromorphology
The macromorphological analysis of S. oleracea and C. bonducella leaves showed that they were smooth, succulent, ovate to triangular or elliptic oblong, and green in hue. Compared to the alternate leaf of S. oleracea, the petiole of the bipinnate leaf of C. bonducella was short and spiky. The roots of W. somnifera are stout, long, woody, tuberous, fleshy, and whitish-brown in color. The E. officinalis fruit had a greenish-yellow color, was smooth, spherical, or globular in shape, and had six vertical bands that were broader (18–25 mm) than longer (15–20 mm) [Figure 1]. Both apexes had conic depressions [Table 1].
|Figure 1: Fruit of Emblica officinalis (a), roots of Withania somnifera (b), Leaves of Caesalpinia bonducella (c), and Spinacia oleracea (d)|
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|Table 1: Botanical description and distribution of selected antianemic plants|
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S. oleracea and C. bonducella leaves possess distinct types of stomata. Anomocytic stomata surrounded by straight-walled epidermal cells were observed in S. oleracea [Figure 2]. Paracytic stomata surrounded by 2–3 wavy-walled epidermal cells were observed in C. bonducella [Figure 3]. Microscopical examination of the transverse sections of both plants shows a single-layered upper epidermis and a lower epidermis covered with a thick cuticle. Beneath the upper epidermis, two to three layers of closely packed, slightly elongated, unbranched palisade cells and several layers of branched cells as a part of the spongy parenchyma were observed. A lignified bicollateral vascular bundle was observed in both S. oleracea and C. bonducella. Thick-walled trichomes were observed in C. bonducella [Figure 4] and [Figure 5].
|Figure 2: Diagnostic structures of powdered Spinacia oleracea leaves, (a) Vascular bundles; (b) Anomocytic stomata; (c) Spongy parenchyma; (d) epidermal cell; (e) spiral vessels|
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|Figure 3: Diagnostic structures of powdered Caesalpinia bonducella leaves. (a) Spiral vessels; (b) thick-walled trichome; (c) lignified fibers; (d) paracytic stomata|
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A transverse section of W. somnifera showed the stratified, nonlignified parenchymatous cell followed by 2–4 diffused rows of cork cambium. Beneath the cork, a few layers of slightly flattened parenchymatous cells as a cortex were observed with lots of calcium oxalate and starch grains. The secondary phloem is separated from lignified wood vessels by cambium. The secondary phloem consists of isodiametric parenchymatous cells, sieve tubes, and companion cells. The secondary xylem was composed of lignified fibers, medullary rays, and parenchyma [Figure 6].
The outer side of the transverse section of the E. officinalis single epidermal layer was present as an epicarp, which was covered with cuticle. Beneath the epidermis was a layer of subepidermal cells, which were wider, tangentially oblong, and occurred parallel to the epidermis. The largest portion of fruit, the mesocarp, consists of several layers of parenchymatous cells. Vascular strands were scattered in the mesocarp. The vascular bundles are varied in size and composed of clusters of xylem elements and a small group of phloem elements. Xylem elements were highly thick-walled. The phloem consisted of fairly large sieve tubes and companion cells [Figure 7].
|Figure 7: Transverse section of Emblica officinalis fruit under low and high magnification|
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Lignified structures were seen in the powdered leaves of S. oleracea and C. bonducella, the root of W. somnifera, and the fruit of E. officinalis. In the leaf powder of S. oleracea and C. bonducella, stomata, pieces of epidermal cells, spiral vessels, and bundles of vascular cells were seen [Figure 2] and [Figure 3]. Prismatic calcium oxalate crystals were predominantly found in the root of W. somnifera. Polygonal or rectangular, thin-walled cork cells, heavily lignified reticulate or bordered pitted vessels, and tracheids were also observed in the root powder of W. somnifera [Figure 8]. Epidermal cells, parenchyma, sclereids, numerous brown, uniformly sized oil droplets, and a pitted vessel attached with parenchyma and fibers were found in the powder of E. officinalis fruit [Figure 9]. Tannins are important parts of this powder microscopic study. They are known for their antioxidant properties and are what protect and keep ascorbic acid from breaking down when it dries in the sun.
|Figure 8: Diagnostic structures of powdered Withania somnifera roots. (a) Pitted and scalariform vessels; (b) microsphenoidal crystals of calcium oxalate; (c) tracheids; (d) cork cell|
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|Figure 9: Diagnostic structures of powdered Emblica officinalis fruit. (a) Fibers; (b) xylem vessels with fibers; (c) xylem vessels; (d) epidermal cells|
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The acid-insoluble ash value of S. oleracea leaf and E. officinalis fruit was higher than the water-soluble ash value, however the acid insoluble ash value of W. somnifera root and C. bonducella leaf was lower than water-soluble ash value. pH value of extracts from selected parts of S. oleracea, W. somnifera, E. officinalis, and C. bonducella was weakly acidic to acidic ranging from 3.2 to 6.2. The alcohol-soluble extractives were largely greater than aqueous soluble extractive of the respective parts of W. somnifera, E. officinalis, and C. bonducella except the leaves of S. oleracea [Table 2].
|Table 2: Physicochemical evaluation of leaves of Spinacia oleracea and Caesalpinia bonducella, Withania somnifera root, and Emblica officinalis fruit|
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Preliminary phytochemical analysis
A preliminary phytochemical screening was done to identify the types of phytoconstituents found in various extracts. This serves as a vital diagnostic standard for separating raw medications from other compounds. Investigation of the phytochemical profiles of the powders from selected parts of S. oleracea, W. somnifera, E. officinalis, and C. bonducella revealed the presence of alkaloids, glycosides, saponins, phytosterols, flavonoids, terpenoids, reducing sugars, phenol, and tannins [Table 3]. A preliminary phytochemical study of the relevant parts of the chosen plants shows that they have both polar and nonpolar phytocomponents.
|Table 3: Phytochemical screening of leaves of Spinacia oleracea and Caesalpinia bonducella, Withania somnifera root, and Emblica officinalis fruit|
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| Discussion|| |
According to the World Health Organization, the macroscopic and microscopic description of a medicinal plant is the first step toward establishing the identity and the degree of purity of such materials and should be carried out before any tests are undertaken. These studies include macromorphology, microscopic studies, physicochemical studies, and preliminary phytochemical studies. S. oleracea possesses ovate to triangular leaves, whereas C. bonducella has ovate to elliptic leaves. S. oleracea possesses a longer petiole, alternate arrangement, and symmetrical base than C. bonducella, which has a short petiole, bipinnate arrangement, and truncate-rotund base. The leaves of S. oleracea and C. bonducella are simple and entire. W. somnifera has a yellowish-brown, smooth outer surface with longitudinal wrinkles, short and uneven fractures, and straight, conical or cylindrical roots. E. officinalis has greenish-yellow, globular hard fruit with no discernible odor and sour fractures, as well as six longitudinal furrows running from top to bottom [Table 1].
Microscopic examination of the lamina of fresh leaves revealed anomocytic stomata surrounded by straight-walled epidermal cells in S. oleracea and paracytic stomata surrounded by 2–3 wavy-walled epidermal cells in C. bonducella. The shape and form of these structural elements are important for the identification of the plants. Furthermore, leaf measurements are used to distinguish between closely related species and adulterants.
Transverse sections of selected parts of S. oleracea, W. somnifera, E. officinalis, and C. bonducella will help identify the drug in fragmented form as well as in whole form. Histological examination of the midrib of S. oleracea revealed the presence of a lignified collateral vascular bundle, which was the same as in C. bonducella except for the availability of trichomes. The main diagnostic features of W. somnifera root are clustered calcium oxalate crystals as well as small, rounded, simple, and compound starch grains in the cortex region; conjoint, collateral, endarch, and closed vascular bundles; lignified fibers; and medullary rays. The fruit of E. officinalis contains an epicarp, mesocarp, and vascular bundles.
The microscopy of powdered fruit of E. officinalis, roots of W. somnifera, and leaves of S. oleracea and C. bonducella revealed the presence of several ergastic cell contents. However, because these traits are found in all plants, they can't be used to tell fake plants or other parts of plants apart.
The aqueous and alcoholic extractives of selected plants vary significantly. The alcohol-soluble extractives of C. bonducella, E. officinalis, and W. somnifera were found to be more than 10 while being less for S. oleracea. Except for S. oleracea, the water extractives of all plants were >10 and higher than the alcoholic extractives. Extractive values are important for determining when drugs have been previously exhausted.
The pH ranged from 3.2 to 6.2 and was acidic to strongly acidic for extracts from S. oleracea, C. bonducella, E. officinalis, and W. somnifera. Traditionally, hydro-alcoholic extracts have been produced for use as pharmaceuticals. Amponsah et al. found that the pH of the extracts was only slightly acidic, with the exception of E. officinalis. This means that there were no irritations or ulcers in the gastrointestinal tract (GIT).
Ash values are crucial for determining the quality and purity of crude plant extracts. The total amount of ash that is soluble in weak acids represents all its constituent parts. Inorganic adulterants such as metallic salts, silica, and crude medicines make up the component of the item that is acid insoluble. In most cases, the impurity or adulteration is represented by an acid-insoluble ash.
During the phytochemical analysis of the plant extracts in question, different phytoconstituents were found, which was in line with previous phytochemical research.,,, Plants' potential for therapeutic application may be due to the secondary metabolites that are present in them.
When used together, the parameters of the relevant parts of S. oleracea, C. bonducella, E. officinalis, and W. somnifera will help identify the whole or powdered crude drugs of each plant without any doubt.
| Conclusion|| |
The current study has laid down pharmacognostic, phytochemical, and physicochemical profiles that can be used for the identification and quality control of leaves of S. oleracea and C. bonducella, roots of W. somnifera, and fruit of E. officinalis. These are crucial for the proper sourcing of these plants for manufacturers, regulators, and researchers of herbal products. The early phytochemical analysis will support future efforts to isolate significant molecules.
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Conflicts of interest
There are no conflicts of interest.
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Mayuri M Thumar,
Lecturer, B. K. Mody Government Pharmacy College, Rajkot - 360 003, Gujarat
Source of Support: None, Conflict of Interest: None
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8], [Figure 9]
[Table 1], [Table 2], [Table 3]