ESTIMATION OF PHYTOCHEMICAL CONSTITUENTS IN THREE PLANTS WIDELY USED FOR TRADITIONAL TREATMENT OF ASTHMA IN MYANMAR

ESTIMATION OF PHYTOCHEMICAL CONSTITUENTS IN THREE PLANTS WIDELY USED FOR TRADITIONAL TREATMENT OF ASTHMA IN MYANMAR

Za Khai Tuang

Department of Botany, University of Kalay, Kalay, Sagaing District, Myanmar.

ABSTRACT: The crude drug powder extracts from three plants Datura metel L., (Solanaceae), Leucas cephalotes (Roth.). Spreng, Piper betel L., (Piperaceae) were screened for estimating phytochemical constituents. Phytochemical analysis of the extracts revealed that glycoside; tannin, saponin, carbohydrate and phenolic compounds are present in Piper and Leucas species. Besides glycoside, tannin, saponin, carbohydrate and phenolic compound, alkaloid is also present in Datura species. The antimicrobial activity of the plant materials and their usefulness in the treatment of asthma may be due to the presence of active constituents like tannins, alkaloids and phenolic. The present study reported that the phytochemical constituents of three plants being widely used for traditional treatment of asthma in Myanmar, collected from Kalay, Sagaing District.

Keywords: Datura, Leucas, Piper, Antimicrobial, Asthma, Phytochemical

INTRODUCTION: Plants are good sources of a wide variety of compounds, such as phenolic, terpenoids, nitrogen, vitamins and secondary metabolites, which possess antioxidant, anti-microbial, anti-inflammatory, antitumor, antimuta-genic, anti-carcinogenic and diuretic activities ¹. Plants have therefore played a significant role in maintaining human health and improving the quality of human life ^{2, 3}. Even today, the World Health Organization assumes that as many as 80 percent of people still rely mainly on traditional therapies, such as herbal medicines ⁴. Datura metel L. has a wide range of traditional applications. Dried powder from the flower of the D. metel is used to treat asthma, chronic bronchitis, and pain in Myanmar. The dried leaves powder or fruit powder is smoked to cure asthma ⁵.

The whole plant is used externally for earache and smoked to relieve spasmodic asthma. Seeds, leaves, and roots of Datura are used for curing insanity, fever with catarrh, diarrhea, skin diseases and cerebral complications ⁶. The main constituent of the Datura plant is a huge number of tropane alkaloids which increased gradually with an increase in the age of the plant ^{7, 8, 9}. Leucas cephalotes (Roth.) Spreng. is a rainy season weed and commonly found ascending in the hilly regions of Myanmar. The flowers are administered in the form of syrup or with honey for curing cough, colds and asthma. It has also been reported that L. cephalotes is used for the treatment of pain, asthma, bronchitis, dyspepsia, inflammation, diarrhea, wounds, jaundice, and fever ¹⁰.

The leaf juice of Piper betel L. is used for antioxidant, anticancer, anti-allergic, fever, cough, fatigue, asthma, to disinfect wounds externally ^{11, 12}.   The leaf powder of P. betel is traditionally known to be helpful in treating various diseases like boils and abscesses, conjunctivitis, bad breath, constipation, itches, headache, mastitis, otorrhoea, etc leucorrhoea, rheumatism, cuts and injuries ¹⁴. The present study intends to estimate the phytochemical constituents in extracts of the above three plants collected in Kalay, Sagaing Region, Myanmar, which are widely used for the traditional treatment of asthma in Myanmar. It was detected that the three plants shared very similar phytochemical constituents to each other.

EXPERIMENTAL:

Collection of Plant Samples: Fresh leaves of D. metel, L. cephalotes, and P. betel were collected from Kalay, Sagaing District, Myanmar.

Sample Preparation: The leaves of collected specimens were carefully washed with sterile water, dried in room temperature, then powdered, and kept in an airtight container for further use.

Materials: Electronic balance, water bath, beaker, test tube, glass tube, and airtight container were used in this study.

Reagents: Dragendroffʼs reagents and Mayerʼs reagents, 10% FeCl₃, Benedictʼs solution, distilled water, 2% NaCl + 1% gelatin, 10% Lead acetate, 1 drop of 5% naphthol+ 5 drops of conc: H₂SO₄ were used.

Test for Alkaloids: 2 g of dried powder sample was boiled with 10ml of dilute hydrochloric acid for 30 min, allowed to cool, then filtered. The filtrate was tested with modified Dragendroffʼs reagent and Mayerʼs reagent. The presence of alkaloids was confirmed by the formation of an orange and cream precipitate.

Test for Glycosides: 2 g of dried powder sample was boiled with distilled water for 30 minutes, allowed to cool then filtered.

The filtrate was tested with a modified 10% lead acetate reagent. The formation of a pale yellow or yellow precipitate indicates the presence of glycosides.

Test for Reducing Sugar: 2 g of dried powder sample was boiled with distilled water for 30 minutes. A few drops of Benedictʼs solution were then added to the filtrate. The solution gives to characteristic brick red color indicates the presence of reducing sugar.

Test for Tannins: 2 g of dried powder sample was heated with ethanol in labeled test tubes. Each solution was allowed to cool then filtered. The filtrate was tested with 1% gelatin and 2% sodium chloride. A characteristic gelly block indicates the presence of tannin.

Test for Saponins: 2 g of dried powder sample was vigorously shaken with distilled water in a test tube for 10 min; the formation of persistent froth foam indicates the presence of saponin.

Test for Carbohydrates: 2 g of dried powder sample was boiled with distilled water for 30 minutes and filtered. The filtrate was treated with 1 drop of 5% naphthol and shaken, and the test tube was inclined at an angle of 45°C. Then added 5 drops of concentrated sulphuric acid slowly along the side of the test tube. A white ring formed between layers showed the presence of carbohydrates.

Test for Phenolic Compounds: 2 g of dried powder sample was boiled with ethanol extract and filtered. The filtrate was treated with a few drops of 10% ferric chloride solution. The formation of blue or green color indicates the presence of a phenolic compound.

RESULTS AND DISCUSSION:

Phytochemical Analysis of Datura metel L.: The crude extract samples of D. metel, commonly called Thorn Apple, and Padaing-phyu in Myanmar name Fig. 1 were analyzed, and the results were tabulated Table 1. The phytochemical analysis of the crude extract indicated glycoside, tannin, saponin, carbohydrate, phenolic compound, and alkaloid were present in Datura species Table 1, Fig. 2.

These compounds are known as biologically active and therefore support antimicrobial activity. Datura leaves have also been used for herbal medicine as a bronchodilator, anesthetic, hallucinogenic, anti-spasmodic ¹⁵, calm cough, burns, treachitis and to treat laryngitis ¹⁶. In Myanmar, dried or fresh leaves were widely used for the traditional treatment of asthma as oral. However, it was also reported that several Datura spp. as potential poisoning due to their hallucinogenic property ¹⁷.

TABLE 1: QUALITATIVE ANALYSIS OF PHYTOCHEMICALS IN D. METEL LEAVES

Fig. 1: (A) Habit of Datura metel L.; (B) Inflorescence of Datura metel L.; (C) Leaves of Datura metel L.; (D) Powder of Datura metel L.

FIG. 2: (A) GLYCOSIDE; (B) PHENOLIC COMPOUND; (C) SAPONIN; (D) TANNIN; (E) ALKALOID; (F) CARBOHYDRATE

Phytochemical Analysis of Leucas cephalotes (Roth.) Spreng: The crude extract samples of guma of L. cephalotes, locally called as Pingu-htaik-peik in Myanmar Fig. 3 were also analyzed, and the results were tabulated in Table 2. Like D. metel, glycoside, tannin, saponin, carbohydrate and phenolic compounds were present in L. cephalotes. However, alkaloids could not detect in L. cephalotes Table 2, Fig. 4. Previous reports indicated that tridecanoic acid, lauric acid, adipic acid, labellenic acid, glutaric acid ¹⁸, oleanolic acid, triterpenes, flavones and sterols ¹⁹ were also found in Leucas species. These compounds are suggestive of their possible antimicrobial properties. As long as it is widely used to treat asthma in Myanmar, it is also reported that Leucas spp. are traditionally employed for the treatment of urinary complaints, fever, skin diseases, liver disorders, cold and cough ²⁰, snake bite, bronchitis, inflammation, dyspepsia, paralysis and leucoma, urinary discharge ²¹, chronic, malaria, asthma, bleeding ²², anti-fungal, antioxidant, anti-pyretic, antinociceptive, anti-microbial, analgesic, etc. ^{10, 23, 24}

TABLE 2: QUALITATIVE ANALYSIS OF PHYTOCHEMICALS IN L. CEPHALOTES LEAVES

FIG. 3: (A) HABIT OF LEUCAS CEPHALOTES (ROTH). SPRENG; (B) INFLORESCENCE OF LEUCAS CEPHALOTES (ROTH) SPRENG; (C) LEAVES OF LEUCAS CEPHALOTES (ROTH). SPRENG; (D) POWDER OF LEUCAS CEPHALOTES (ROTH) SPRENG

FIG. 4: (A) GLYCOSIDE; (B) TANNIN; (C) SAPONIN; (D) PHENOLIC COMPOUND; (E) CARBOHYDRATE

Phytochemical Analysis of Piper betle L.: A common name betel of locally called Kun in Myanmar of P. betle Fig. 5 is widely used for the treatment of various ailments since century ^{25, 11}. The crude extract of P. betle was also studied in the current work, and the phytochemical analysis results were tabulated in Table 3, Fig. 6. As shown in Table 3, the present study detected that glycosides, tannins, and saponins compounds were present in betel leaves. Additionally, it was reported that the betel leaf contains minerals, protein, carbohydrates, fats, fibers, water, essential oils ²⁶, alkaloids, different vitamins like vitamin C, nicotinic acid, vitamin A, thiamine, riboflavin, calcium, iron, and iodine ¹³. It has also been suggested that the presence of phenolic natural compound flavonoids in the plant extracts showing antioxidant activity ^27-29. Hence, the phenolic compounds identified in the betel leaves extract might contribute to the antioxidant activity ²⁷. Betel is also believed that a blessed as an evergreen and perennial plant that God has given the shape of his own heart ¹⁴. Besides used as an asthma treatment, betel leaves are also used for curing eye injury, baby lotion for the newborn, coughs ³⁰, constipation ³¹, digestive and pancreatic lipase stimulant activities ^{30, 32}, bad breath ³⁰, conjunctivitis, headache, itches, mastoiditis, mastitis, otorrhoea, leucorrhoea, rheumatism ³³, bronchitis and dyspnea ³⁴, respiratory catarrhs and antiseptic ³⁵, antibacterial ³⁶, anti-acetylcholinesterase ²⁶, inhibit male reproductive competence ³⁷, suggested to against Covid-19 due to the presence of aurantiamide ³⁸, and also used to improve meat cholesterol levels of Bali duck with fermented rice husk ³⁹.

FIG. 5: (A) HABIT OF PIPER BETLE L.; (B) LEAVES OF PIPER BETLE L.; (C) POWDER OF PIPER BETLE L.

FIG. 6: (A) GLYCOSIDE; (B) PHENOLIC COMPOUND; (C) SAPONIN; (D) TANNIN; (E) CARBOHYDRATE

TABLE 3: QUALITATIVE ANALYSIS OF PHYTOCHEMICALS IN PIPER BETLE L. LEAVES

CONCLUSION: From the above studies, it was found that the three plants widely used to treat asthma as a traditional medicine in Myanmar shared very close phytochemical constituents Table 4. It is concluded that traditional plants represent sources of biologically active components and antimicrobial with stable that can establish a scientific base for the use of plants in modern medicine ^{10, 40-42}. Hence, it is hoped that scientifically evaluated ethnomedical preparation can be extended for future exploration into the field of pharmacology and other biological actions for drug discovery.

TABLE 4: COMPARISON OF PHYTOCHEMICAL CONSTITUENTS BETWEEN THE THREE PLANTS

+ = positive; - = negative

ACKNOWLEDGEMENT: I gratefully acknow-ledge Professor Khin Thida Soe for allowing me to do this work in the Department of Botany, University of Kalay.

Funding: This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

REFERENCES:

1. Wadikar DD and Patki PE: Coleus aromaticus: a therapeutic herb with multiple potentials. J Food Sci Technol 2016; 53(7): 2895-2901.
2. Aryani, Suprayitno E, Sasmito BB and Hardoko: Characterization and identification of active compound of ocellated snakehead (Channa pleurophthalmus blkr) waste charcoal potentialas antiallergy drug. Int J Pharm Sci & Res 2021; 12(2): 794-02.
3. Tamilselvan N, Thirumalai T, Elumalai E, Balaji R and David E: Pharmacognosy of Coccinia grandis: A review. Asian Pac J Trop Biomed 2011; 1(2): S299-S302.
4. Yebpella GG, Adeyemi Hassan MM, Hammuel C, Magomya AM, Agbaji AS and Okonkwo EM: Phtyochemical screening and comparative study of antimicrobial activity of Aloe vera various extracts. African J Microbiol Res 2011; 5(10): 1182-87.
5. Rahmatullah M, Mollik MA, Islam MK, Islam MR, Jahan FI, Khatun Z, Seraj S, Chowdhury MH, Islam F, Miajee ZU and Jahan R: A survey of medicinal and functional food plants used by the folk medicinal practitioners of three villages in Sreepur Upazilla, Magura district, Bangladesh. Am J Sustain Agric 2010; 4(3): 363-73.
6. Abad-Santos F, Novalbos J, Gallego-Sandín J and Gálvez-Múgica MA: Regulación del tono bronquial en la enfermedad pulmonar obstructiva crónica (EPOC): Papel de los receptores muscarínicos. An Med Interna 2003; 20(4): 201-05.
7. Benítez G, March-Salas M, Villa-Kamel A, Cháves-Jiménez U, Hernández J, Montes-Osuna N, Moreno-Chocano J, Cariñanos P: The genus Datura (Solanaceae) in Mexico and Spain – Ethnobotanical perspective at the interface of medical and illicit uses. J Ethnopharmacol 2017; 219(12): 133-51.
8. Nargish-Firdaus MHK and Uzma-Viquar: Potential and pharmacological actions of dhatura safed (Datura metel): as a deadly poison and as a drug: an overview. IJPSR 2020; 11(7): 2320-5148.
9. An P and Issn J: Phytochemical profile, antibacterial and antidiabetic effects of crude aqueous leaf extract of datura stramonium. Pharmacophore 2014; 5(2): 273-78.
10. Rahman SMM, Mony T, Ahammed K, Naher S, Haque R and Jui SM: Qualitative phytochemical screening and evaluation of analgesic and antidiarrheal activity of ethanolic extract of Leucas cephalotes J Pharmacogn Phytochem 2018; 7(5): 1484-92.
11. Chowdhury S and Chakraborty P: Antioxidant activity of eugenol in Piper betel leaf extract. J Fam Med Prim Care 2020; 9(1): 327-31.
12. Noor A, Gunasekaran S and Vijayalakshmi MA: Efficacy of salivary and diastase extracts of Piper betle in modulating the cellular stress in placental trophoblast during preeclampsia. Pharmacog Res 2018; 10(10): 24-3.
13. Periyanayagam K, Jagadeesan M, Kavimani S and Vetriselvan T: Pharmacognostical and Phyto-physicochemical profile of the leaves of Piper betle var Pachaikodi (Piperaceae) valuable assessment of its quality. Asian Pac J Trop Biomed 2012; 2(2): S506–S510.
14. Pradhan K, Suri A, Pradhan DK and Biswasroy P: Golden heart of the nature : Piper betle J Pharmacogn Phytochem 2013; 1(6): 147-67.
15. Dabur R, Ali M, Singh H, Gupta J and Sharma GL: A novel antifungal pyrrole derivative from Datura metel Pharmazie 2004; 59(7): 568-70.
16. Partap M, Gupta RC and Pradhan SK: Comparative analysis of morphology and phytochemical constituents in different populations and morphotypes of Datura innoxia and Datura metel L. from punjab plains. Asian J Pharm Clin Res 2019; 12(1): 193.
17. Kerchner A and Farkas A: Worldwide poisoning potential of Brugmansia and Datura. Forensic Toxicol 2020; 38(1): 30-41.
18. Bahadur KD and Sen AB: Chemical examination of Leucas cephalotes. Pharm Biol 1968; 2(140): 1453-54.
19. Miyaichi Y, Segawa A and Tomimori T: Studies on nepalese crude drugs (xxix) chemical constituents of dronapuspi, the whole herb of Leucas cephalotes Spreng Chem Pharm Bull 2006; 54(10): 1370-79.
20. Khan AV, Uddin Ahmed Q, Khan AA and Shukla I: In-vitro antibacterial efficacy of Leucas cephalotes (Roth) Spreng. (Lamiaceae) against some gram-positive and gram-negative human pathogens. Int J Agric Food Res 2014; 3(3): 1-9.
21. DeFilipps RA and Krupnick GA: The medicinal plants of Myanmar. PhytoKeys 2018; 341(102): 1-341.
22. Khare CP: Indian Medicinal Plants. Indian Med Plants 2007; 1-1.
23. Agnes K, Nirmala and Kanchana M: Leucas aspera – A review of its biological activity. Syst Rev Pharm 2018; 9(1): 41-44.
24. Nidhal N, Zhou XM, Chen G, Zhang B, Han C and Song X: Chemical constituents of Leucas zeylanica and their chemotaxonomic significance. Biochem Syst Ecol 2019; 89(10): 104006.
25. Adawiyah-Umar R, Sanusi A, Nizam-Zahary M, Rohin MAK and Ismail S: Chemical composition and the potential biological activities of Piper betel- a review. Malaysian J Appl Sci 2018; 3(1): 1-8.
26. Karak S, Acharya J, Begum S, Mazumdar I, Kundu R and De B: Essential oil of Piper betle leaves: chemical composition, anti-acetylcholinesterase, anti-β-glucuronidase and cytotoxic properties. J Appl Res Med Aromat Plants 2018; 10(6): 85-92.
27. Madhumita M, Guha P and Nag A: Extraction of betel leaves (Piper betle) essential oil and its bio-actives identification: Process optimization, GC-MS analysis and anti-microbial activity. Ind Crops Prod 2019; 138(7): 111578.
28. Sarma C, Rasane P, Kaur S, Singh J, Singh J, Gat Y, Garba U, Kaur D and Dhawan K: Antioxidant and antimicrobial potential of selected varieties of Piper betle (Betel leaf). An Acad Bras Cienc 2018; 90(4): 3871-78.
29. Lourenço SC, Moldão-Martins M and Alves VD: Antioxidants of natural plant origins: from sources to food industry applications. Molecules 2019; 24(22): 14-16.
30. Madhumita M, Guha P and Nag A: Bio-actives of betel leaf (Piper betle): a comprehensive review on extraction, isolation, characterization, and biological activity. Phyther Res 2020; 34(10): 2609-27.
31. Punjani BL and Kumar V: Traditional medicinal plant remedies to treat cough and asthmatic disorders in the Aravalli ranges in North Gujarat, India. J Nat Remedies 2002; 2(2): 173-78.
32. Rawat AKS, Tripathi RD, Khan AJ and Balasubrahmanyam VR: Essential oil components as markers for identification of Piper betle cultivars. Biochem Syst Ecol 1989; 17(1): 35-38.
33. Agarwal T, Singh R, Shukla AD, Waris I and Gujrati A: Comparative analysis of antibacterial activity of four Piper betel Adv Appl Sci Res 2012; 3(2): 698-705.
34. Mula S, Banerjee D, Patro BS, Bhattacharya S, Barik A, Bandyopadhyay SK and Chattopadhyay S: Inhibitory property of the Piper betel phenolics against photosensitization-induced biological damages. Bioorganic Med Chem 2008; 16(6): 2932-38.
35. Amalia H, Sitompul R, Hutauruk J and Mun A: Effectiveness of Piper betle leaf infusion as a palpebral skin antiseptic. Universa Med 2009; 28(2): 83-91.
36. Roy A and Guha A: Formulation and characterization of Betel leaf (Piper betle) essential oil based nanoemulsion and its in vitro antibacterial efficacy against selected food pathogens. J Food Process Preserv 2018; 42(6): 1-7.
37. Arawwawala LDAM, Arambewela LSR and Ratnasooriya WD: Gastroprotective effect of Piper betle leaves grown in Sri Lanka. J Ayurveda Integ Med 2014; 5(1): 38.
38. Sengupta P: Use of Piper betel to combat COVID19. Prepare@U. 2020; 1(1): 1–10.
39. Partama IBG, Yadnya TGB, Trisnadewi AAAS and Sukada IK: Fermented rice husk utilization of effective microorganisms-4 supplemented with Piper betle L. performance, meat quality, antioxidant capacity, and meat cholesterol levels of Bali duck. Int J life Sci 2018; 2(3): 98-110.
40. Caesar LK, Kellogg JJ, Kvalheim OM and Cech NB: Opportunities and limitations for untargeted mass spectrometry metabolomics to identify biologically active constituents in complex natural product mixtures. J Nat Prod 2019; 82(3): 469-84.
41. Salehi B, Ata A, Kumar NVA, Sharopov F, Ramírez-Alarcón K, Ruiz-Ortega A, Abdulmajid-Ayatollahi S, Fokou PVT, Kobarfard F, Zakaria ZA and Iriti M: Antidiabetic potential of medicinal plants and their active components. Biomolecules 2019; 9(10): 1-111.
42. Durazzo A, D’Addezio L, Camilli E, Piccinelli R, Turrini A, Marletta L, Lucarini M, Lisciani S, Gabrielli P and Gambelli L: From plant compounds to botanicals and back: A current snapshot. Molecules 2018; 23(8): 1844.
    

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    How to cite this article:

    Tuang ZK: Estimation of phytochemical constituents in three plants widely used for traditional treatment of asthma in Myanmar. Int J Pharm Sci & Res 2021; 12(9): 5102-09. doi: 10.13040/IJPSR.0975-8232.12(9).5102-09.

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