|Year : 2022 | Volume
| Issue : 3 | Page : 155-163
Pharmacognostical and phytochemical standardization on stem bark of Mahonia nepalensis DC.: An extrapharmacopoeial plant
Kiran Paudel, Aku Ramamurthy, Gaurav Sharma
Post Graduate Department of Dravyaguna Vigyana, National Institute of Ayurveda, Jaipur, Rajasthan, India
|Date of Submission||26-Jun-2022|
|Date of Acceptance||04-Jul-2022|
|Date of Web Publication||30-Sep-2022|
Department of Dravya Guna Vigyanan, National Institute of Ayurveda, Jaipur 302002, Rajasthan
Source of Support: None, Conflict of Interest: None
Introduction: The tribal people cure hepatic-related disorders with the stem bark of Mahonia nepalensis DC. Aim: The aim of the study was to do Investigation of the Microscopic and Macroscopic Characteristics, Pharmacognostical, Physiochemical Parameters, and Thin Layer Chromatography of Stem Bark of Mahonia nepalensis DC. Materials and Methods: Mahonia nepalensis DC. were collected from natural habitat and authenticated by the proper plant authentication centers. The cell composition of TS slices was examined after they had been dyed and mounted. The powders of the sample drug were studied organoleptically. The Extractive values and Ash values were represented in Pie-chart form designed by using Microsoft Excel. Results: The Bark of the Mahonia nepalensis DC. is brown in color externally but the inner side is yellow with features of general anatomy of dicot stem on transverse section. The pharmacognosy and phytochemical investigations of the stem bark of Mahonia nepalensis DC shows presence of starch grain, intermedullary rays as diagnostic characters. Mayer’s reagent test and Dragon Droff’s reagent test are positive in the test signifies that the present of Alkaloids and TLC values were evaluated. Conclusion: The key diagnostic characteristics of Mahonia nepalensis DC’s stem were identified from its pharmacognosy. The stem of Mahonia nepalensis DC was authenticated and subjected to quality control using phytochemical tests and TLC, which were derived and described.
Keywords: Mahonia nepalensis DC, pharmcognostical study, phytochemical study
|How to cite this article:|
Paudel K, Ramamurthy A, Sharma G. Pharmacognostical and phytochemical standardization on stem bark of Mahonia nepalensis DC.: An extrapharmacopoeial plant. J Indian Sys Medicine 2022;10:155-63
|How to cite this URL:|
Paudel K, Ramamurthy A, Sharma G. Pharmacognostical and phytochemical standardization on stem bark of Mahonia nepalensis DC.: An extrapharmacopoeial plant. J Indian Sys Medicine [serial online] 2022 [cited 2022 Nov 28];10:155-63. Available from: https://www.joinsysmed.com/text.asp?2022/10/3/155/357687
| Introduction|| |
Pharmacognostical study is defined as “A branch of science that aims at knowledge of the morphological nature and the structure, Macro and Microscopic Examination with active constituents present in crude drugs”. Pharmacognosy is an important bridge between the pharmaceutical and basic sciences. Pharmcognosy is a vital link between Ayurvedic and Allopathic system of Medicine., Extrapharmacopoeial plants are those plants which are not recorded in the different types of Ayurveda treaties but predominantly used in the general practice by the Traditional healers. In Ayurveda it is commonly known as Anukta Dravya.Mahonia nepalensis DC is Anukta Dravya of Ayurveda belongs to family Berberidaceae.Mahonia nepalensis DC. is a 1.2–3 m tall evergreen shrub with sparsely branched erect stems up to 20 cm in diameter. Bark is pale brown with rough and croky looks from outside. From the inner side it has bright yellow in color. The leaves are present at the terminal of twigs which is pinnately compound and length is in between 18 to 45 cm long. Leaflets are usually 7 to 17 in numbers and the length of each leaflet varies from 3.8 to 10 cm long. The shape of leaflets are ovate or lanceolate or acuminate at the base of lateral pairs and the leaflet have 3 to 5 basal nerve prominent from behind and from above it is shining greenish color. Flowers at the tops of the branches are in thick, tall racemes that are 5–12.5 cm long and fascicled with yellow, 5–6.5 mm long, sweet-scented flowers. Pedicles is 1.25 to 4 mm long. Berries are oval and 5–10 mm long. They are blue-black and glaucous. Distribution: Temperate Himalaya, 4000–8000 ft. from Garhwal to Bhutan, Khasia Hills 4000–5000 ft., Mergui, Nilgiri Mts. 5000–8000 ft. It is used by the traditional Practitioners in different forms for the treatment purpose. Generally, Mahonia bark is used as Antidysenteric, Antidiarrheic drug. Berries are diuretic and demulcent in dysentery. The berries are considered diuretic, and demulcent in dysentery. In Garhwal it is commonly known as Gurm, In Haldia it is known as Jaunsar and Khoru; In Nepal the plant is known as Jamanemandro, Milkissi and in Punjab it is known as Amudanda. Ethnobotanically, Mahonia plant is used for the diseases related to Eye, Gastrointestinal and Hepatic related disorders. Phytochemical test of the plant shows the presence of terpenoid, cardiac glycoside, quinines and steroid. Bisbenzylisoquinolines homoaromoline and isotereandrine are found in all type of stem extracts. Berberine, Oxycanthine, Epiberberine, Columbamine, Berbamine, Dihydrokarachine, Karachine are also present in stem of M. nepalensis DC. Hence, present study is related to the Pharmacological standardization of Mahonia plant with its anatomical exploration and phytochemical investigations.
| Material and Methods|| |
Collection and Authentication of Plant
The Stem and the bark of Plant were collected from its Natural habitat of Bosan Forest, Chalnakhel and Phulchoki, Goadawari region of Kathmandu Valley, Nepal in February, 2020 and March, 2021 by first Author. The Plant was identified as Mahonia nepalensis (DC.) with Identification Berberidaceae Family by expert taxonomists. [Figure 1] and [Figure 2]
|Figure 1: (A) Plant at natural habitat. (B) Bark removing of Plant for Experimental trial. (C) Leaf of the Plant showing Compound leaflets. (D) The Shrub having Inflorescences. (E) Cultivation of Plant in Botanical Garden. (F) Twig with Inflorescence|
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|Figure 2: Plant with Inflorescence and Plant prepared for Herbarium Sample for Plant Authentication|
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PLANT AUTHENTICATION: For Mahonia nepalensis DC.- NIScPR/RHMD/Consult/2021/3837-38-1.
Formalin, Chloral Hydrate, Glycerin, Crystal Violet, Safranin, Methylene Blue, Iodine Stain, Acetic Acid, Ethyl Alcohol, Toludine blue and all other chemicals as required for the Phytochemical Study and Pharmacognostical standardization.,
Pharmacognostical Standardization Parameters
Both the powdered samples of the drug and the collected samples of Herbarium were examined under a microscope and with the naked eye, respectively. Size, Shape, Color, Appearance and Odor of the powder sample were studied Organoleptically.
The stem of Mahonia nepalensis DC were cut into transverse sections using a razor and the sections were then exposed to the chemical for a short period of time. The best section was chosen, mounted temporarily with glycerin, and examined under a light microscope.
For Powder microscopy, powder is obtained by making powder of the dried cut pieces of the mature stem bark. The small pieces of stem bark were firstly grounded in the Grinder machine and then Pulverized for making the fine Powder. Powder of Mahonia was treated with different chemical reagents.
According to standard methods, the Mahonia’s Physiochemical Values including moisture content, extractive values, and ash values were determined.
Following standard methods, fresh extract were checked for to detect the varieties of active phytochemicals, including phenols, flavonoids, proteins, reducing sugar, tannin, carbohydrates, lipids, saponin, triterpenoid alkaloid, resins and volatile oils.
It was carried out using a “Thin Layer Chromatography (TLC) plate coated with a 0.25 mm layer of silica gel 60 F254” and a fluorescent indicator (Each plate measures 2 cm in width and 10 cm in length.). The Sample spot were made in activated TLC Plate and Plate was kept in the prepared mobile solution and finally plate was visualized under UV 366 nm with Iodine Reagent.,
| Results|| |
Macroscopy of Stem
Stem of Mahonia nepalensis is Greenish is Young and brown in Mature from outside and from inner side of the stem it is yellow in color. Stem is erect, subsimple with leafy at top only. Stem are terete, angled or sulcate. Internode is 15–20 cm long. In Mahonia nepalensis, according to the age of plant and exposure to the environment it varies from green to Dark brown as well. Stem doesn’t contain any spine like structure but the leaflets have sharpy spionus thoothed.
Macroscopy of Stem Bark
The stem bark of Mahonia nepalensis DC. is brownish from outside and Yellowish from inside. Stem bark is 4–8 mm in thickness. Young stem bark is greenish with short node and Mature have long Internode having small stem at node. At dried condition inner surface of Stem bark is Black and outer surface is Brown, 15–20 longitudinal alternative ridge and furrow are present in adult stem bark [Figure 3].
|Figure 3: (A) Yellow color Stem Bark of Mahonia nepalensis DC. (B) Dried Stem Bark. (C) Powdered Stem Bark|
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Transverse section of stem
The cork of the mature stem is composed of 20–30 layers of thick-walled cells. A narrow band of phelloderm, only 4–6 layers wide, follows, consisting of thin-walled parenchymas that are cut by large, funnel-shaped medullary rays. Sieve tubes, sieve plates, companion cells, phloem parenchyma and phloem fibers are the components of the secondary phloem. The phloem fibers are lengthy, have thick, lignified walls, a large lumen, and primarily pointy tapering ends. There are also some sclerides with thick lignified walls in the secondary phloem area. Tracheids, xylem fibers and xylem parenchyma make up the secondary xylem. Most vessels have pit borders. The walls of the short tracheids are simply pitted. The xylem fibers seem to be concentrated in the lower portion of each individual vascular bundle that resembles a letter V in transverse section. The xylem parenchyma has simple holes and thick walls that range in shape from square to polygonal. The cells have thick, pitted walls and range in shape from polygonal to circular to isodiametric [Figure 4].
|Figure 4: (A) Stem of Mahonia nepalensis DC. for TS and LS. (B) Transverse section of stem Seen in Stereoscope (X10). (C) Longitudinal section showing Parenchymal cells. (D) Parenchymal cells. (E) Transverse section showing Medullary rays (X40). (F) Detailed structure of Xylem and Phloem at X40. (G) Medullary Rays, Xylem and Pericyclic fibers at X10. (H) Cork, Medullary rays at X10. (I) Phloem and Medullary rays at X40|
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The Powder sample of Mahonia was examined through organoleptically. The powder is Yellow in color with minute Black Particles present in it. The odor is Peculiar and the bitter in taste and fibrous rough in texture. The Consistency was Solid form.
The stem powder has a yellowish in color which is made up of fibrous material and has a bitter flavor. It displays the fibers and crystals that are present. The fibers are lengthy, lignified, have a tiny lumen and occasionally have forked ends. The crystals of calcium oxalate resemble needles like appearances and also notably observed that the vessels are present with bordered pits and thickening in the spiral pattern [Figure 5].
|Figure 5: (A) Preparation of Powder for Powder Microscopy. (B) Xylem X40. (C) Phloem X40. (D) Calcium Oxalate crystal X40. (E) Parenchymal cells X40. (F) Crystals at X40. (G) Pericyclic fibers X40. (H) Paranchymal cells bundle X40. (I) Starch grain stained with Iodine at X40|
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All physiochemical parameters were calculated and the results are shown using various pie-chart methods. The pH Value of an aqueous liquid may be defined as “The common reciprocal of Hydrogen ion concentration expressed in gram per liter”. The pH is measured with the help of digital pH meter. The pH Value of the Mahonia nepalensis DC is 6.4. Alcohol Soluble Extractive Value of 5 gm air-dried, crushed Mahonia powder is extracted from Rotary Extraction method with alcohol solvent and sample was filtrate and then dried on Hot water bath. Water soluble extractive value was calculated from 5 gm air-dried, Powdered drug in the Rotary extractor and filtrate sample was dried in hot water bath. Petroleum ether soluble extraction was done in Soxhlet Extractor for 6 hours. Filtrated extract was dried over Water Bath and then Extract was obtained. The alcohol soluble extractive, Water soluble, Ether soluble extractive values are 2.24%, 5.91% and 0.176% respectively. The Total Ash Value of 5 gm Mahonia powder was obtained by ignition at 450°C at Muffle furnace. Acid Insoluble Ash value was obtained by boiling total ash with 25 ml of 2M HCl for 5 min and then filtrate paper was ignited and cool in a desiccators and weight. Water soluble ash was calculated by boiling total ash for 5 minutes with 25 ml of water. The ignited filter paper was cooled and weighted. The total ash, acid-insoluble ash and water soluble ash of stem are 2.35%, 0.717%, 0.8143% respectively. Parameters for Physiochemical study are given in Pie-Chart representation [Figure 6].
Freshly made extracts were examined for the presence of phenols, flavonoids, proteins, reducing sugar, carbohydrates, lipids, tannin, saponin, triterpenoid alkaloid, resins and volatile oils, among other active phytocompounds. [Table 1][Table 2][Table 3][Table 4][Table 5][Table 6][Table 7]
In activated thin layer chromatography plate alcoholic extract of stem bark of Mahonia nepalensis was absorbed by capillary. Mobile solution was prepared from Toluene: Water: Glycial acetic acid ratio of 6:3:1. After development of Spots in the TLC Plate was sprayed with Iodine and was visualized under UV 366 nm. Rf Value of the calculated by “Dividing the distance travelled by the spots by the distance travelled by the front of the mobile phase”. The Rf Value is tabulated in [Figure 7] and [Figure 8].
|Figure 7: Sample of Mahonia nepalensis DC Showing Distance of Solvent and Spots in the TLC Plate with calculated Rf Values|
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|Figure 8: Graph Showing Rf Values with distance of Spot when solvent move distance of 6 cm|
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| Discussion|| |
Mahonia nepalensis DC is a woody shrub with compound leaf with leaflet bearing Spinous thorny like appearances. The Key point for the identification of Mahonia nepalensis is that the plant have 4–6 (-8) leaflets with height of 2–3 meters. Flowering time of plant is Jan, Feb and April. Habitat of the Plant is Warm Temperate forest of altitude 1200–1900 m. This Keys point for identification of plant can help to identify plant in its natural habitat. The stem bark is brown in color with longitudinal furrow but in the young stem is greenish color whereas internally, it is yellow in color with thick bark. Even though it is from berberidaceae family, it doesn’t bear any thorn like structure in stem. Microscopic character of the stem such as presence of crystals, xylem, medullary rays and phloem are important characters for identification of diacot plant. Microscopic features of TS of stem shows tannin inter xylary phloem which is special character of Mahonia nepalensis DC. The Powder of M. nepalensis stem appeared to be yellow, fibers with bitter taste. The grade and purity of the crude drug can be ascertained by evaluating the ash value. The qualitative and quantitative evaluation of crude drugs can benefit from extractive value analyses. Total Ash content was 2.35% indicating fewer amounts of inorganic components are present. Minerals components are very less in the water soluble ash having value of 0.8143%. The siliceous component is very low as seen by the 0.717% acid-insoluble ash. “Alcohol soluble extractive” and “Water soluble extractive” of 2.24% and 5.91% suggestive of presence only a few amount of some examples of polar secondary metabolites such as steroids, triterpenoides, coumarins, phenols, alkaloids, glycosides and tannins. Fewer lower polar chemicals were present showed by the less amount of petroleum ether extracts value as 0.176%. Phytochemical test shows that the present of carbohydrate through tests of Molish’s test, Benedict test, Fehling test. Present of alkaloids is done by Dragendroff test, Present of Amino acid through Ninhydrine test, Present of Protein through Biuret test, Millon test. Since, Mahonia nepalensis DC. is Extrapharmacoepial plant, in API this plant hasn’t been mentioned till now for the comparative study of pharmacognostical and phytochemical values. The TLC shows that the Mahonia nepalensis DC have a yellow colored spots in the TLC plate with different Rf values. The bands in the sample are obtained at Rfs 0.96, 0.58, 0.55, 0.53, 0.5, 0.46, 0.43, 0.35, 0.31 and 0.2. The largest spot was identified at Rf 0.43.
| Conclusion|| |
Mahonia nepalensis DC. is a woody shrub having compound leaflet of height 2–3 meters plant of habitat of the plant is Warm Temperate forest of altitude 1200–1900 m. Typical character of the stem is of abundant Starch. Present of yellow color stem with interxylary pitting helps for identification of the plant. Leaflets are 4–6 (-8) in number having spinous thorny like structure is the Key point for identification of Mahonia nepalensis DC. Pharmacognostical and Phytochemical standardization of the plant helps to further identification of Chemical compound present in it. The plant is studied only in the Experimental trials till now. Mahonia is widely used in Ethnobotanical purpose. So, the plant should study in the clinical trial point of view on the basis of ethnomedicinal value related to plant.
We would like to thank to the curators of KATH- Suvash Khatri, Head, Chief Scientist and Ganga Dutt Bhatta, Chief Scientist and TUCH Curators. We are thankful to Dr. Sunita Garg, Former Chief Scientist, RHMD, CSIR-NIScPR, New Delhi. We are grateful to Dr. Mukti Poudeyal, PhD and Dr. Dharma Raj Adhikari for their expertise on location and identification of Plant. We are thankful to Raghunath Yadav Sir for proper guidance during Laboratory investigations.
Financial Support and Sponsorship
Conflicts of Interest
There are no conflicts of interest.
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[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6], [Table 7]