GC-MS METABOLITE PROFILING OF BIOACTIVE COMPOUNDS IN LEAF, FRUIT AND SEED EXTRACTS OF SYZYZIUM CUMINI (L.)HTML Full Text
GC-MS METABOLITE PROFILING OF BIOACTIVE COMPOUNDS IN LEAF, FRUIT AND SEED EXTRACTS OF SYZYZIUM CUMINI (L.)
Satyavathi Chilumula * 1, Jyothi Chaitanya Pagadala 2, Madhavi Bramhanapalli 2 and Lakshmi Bhavani Nelavelli 1
Department of Botany 1, University College of Science, Saifabad, Osmania University, Hyderabad - 500004, Telangana, India.
Department of Plant Sciences 2, School of Life Sciences, University of Hyderabad, Gachibowli, Hyderabad - 500046, Telangana, India.
ABSTRACT: The present study aimed to explore the primary metabolites present in leaf, fruit, and seed crude extracts of Syzygium cumini L by Gas chromatography - Mass spectrometry. The compounds in the leaf, seed, and fruit extracts resulted from the GC-MS analysis was relevant to the National Institute of Standards and Technology (NIST) library. The metabolite profiling of the extracts revealed the presence of carboxylic acid and fatty acid found to be common in leaf, fruit, and seed extracts. And metabolites 6-oxa-3-thiaoctanoic acid, bisoprolol, phenacetin, tropin, alizarin, gonodoic acid are present in seeds that are not found in leaves and fruit metabolite profile. In contrast, Tetra decanoic acid, Azelaic acid, Penicillamine and S (-)-Cathinone are found only in leaves. Leaf, fruit and seed extracts of Syzyzium cumini L contains various bioactive compounds and consequently has a variety of medicinal properties for the treatment of different diseases like thyroid cancer, heart diseases, kidney stones and rheumatoid arthritis. Some metabolites found to be active as antidiabetic, anti-inflammatory, anti-cancerous, antioxidant, anti-bacterial and antifungal activities identified in leaf, fruit and seed extracts of Syzyzium cumini L. This study may pave the way for the production of herbal medicines for various ailments using leaf, fruit, and seed extracts of Syzyzium cumini L.
Syzyzium cumini L, GC-MS (Gas chromatography - Mass spectrometry), metabolite profiling, Leaf, fruit and seed extracts
INTRODUCTION: In the olden days, people mostly used herbal medicines to treat all diseases. Treating diseases through natural medicine is the earlier method that is known to humankind. But currently proper biological study is needed to find novel drugs also discovery supplementary medicinal properties of the plants for medicinal care 1.
There are undefined medicinal properties of many medicinal plants yet to explore. In India, the medicinal plants using by Ayurveda, Siddha, Homeopathy, etc., for various ailments 2. 80% of the world’s population (WHO) based on traditional plant-based medicines for primary healthcare needs. Recently much of the research was being carried out in medicinal plants 3, 4.
Because the synthetic drugs which were used by human have many side effects that often lead to serious complications 5. And the development of herbal medicine done by the primary screening of the compounds in the plant extracts 6, 7. In modern or synthetic medicine, herbal medicine was the lifesaving drug 8, 9 due to its usual mystic powers as well as for medicinal merits and virtues and Around many plant drugs for which synthetic ones are quiet absent 8, 9. Only 6% of medicinal plant species studied for their biological activity and only a few have phytochemically investigated. So there is a need to study the metabolite profiling of medicinal plants 10.
Syzygium cumini (Syn. Eugenia jambolana) belongs to the Myrtaceae family. Fruits and seeds of Syzygium cumini (Jamun) have hypoglycaemic, anti-inflammatory, antipyretic, hypolipidemic, and antioxidant activities 11.
Several phytochemicals identified in Syzygium cumini (Jamun) fruits, leaves, and seeds some of which include: alkaloids, saponins, flavonoids, tannins, glycosides, anthraquinones etc. and also phyto-chemicals of fruits, leaves, and seeds are non-nutritive chemicals of plants and possess disease preventive or protective properties. These include anticancer activity, antibacterial, antioxidant, hormonal action, enzyme stimulation, and many others 12.
GC-MS is a powerful technique used for the specific detection and potential identification of compounds based on the molecular mass in a complex mixture 13. In a typical MS procedure, a sample is ionized and separated according to their mass-to-charge ratio. Ions of the same mass-to-charge ratio will undergo the same amount of deflection 14.
The atoms or molecules in the sample can be identified by correlating known masses to the identified masses or through a characteristic fragmentation pattern 15. In recent period GC-MS studies have been increasingly applied for the analysis of most of the medicinal plants 16.
This technique has proved to be a valuable method for the analysis of non-polar components and volatile essential oils, fatty acids, and lipids present in plants as secondary metabolites 17, 18. All these medicinal properties of Syzygium cumini (Jamun) fruits, leaves, and seeds are due to the presence of its phytochemical constituents, which not yet explored thoroughly. So, in this study, the Gas Chromatogram Mass Spectrometric method (GCMS) was used to carry out metabolite profiling of the crude extracts of dried Syzygium cumini (Jamun) fruits, leaves, and seeds.
MATERIALS AND METHODS: One mg of the finely powdered leaf, fruit, and seeds powder is added to 60 ml ribitol in Milli-Q water along with 20 ml of methoxy pyridine solution in a separate test tubes. The mixture incubated for 90 min at 70 ºC and supernatant was collected. Then the supernatant sample solution is added to 40 ml of N trimethylsilyl-N-methyl tri-fluoroacetamide. This sample mixture incubated at 70 ºC for 30 min followed by centrifugation at 12000 rpm for 15 minutes.
These samples then analyzed by LECO-PEGATUS GCXGC-TOF-MS system (LECO Corporation, USA) equipped with a 30m Rxi-5ms column with 0.25 internal diameter and 0.25µm film thickness (Restek, USA).The injection temperature, interface, ion source set at 250 ºC, 225 ºC, 200 ºC respectively for the samples.
For separation of metabolites according to their groups, the run program was set at isothermal heating at 70 ºC program for 5 min, followed by 5 ºC of minimum oven temperature and final heating at 290 ºC for 5 min and results are observed. The separation of compounds results in more than one peak for a metabolite of interest. Due to partial silylation or isomerization of methoxy animated compounds such as sugars 19.
Results: The metabolite profiling of the leaf, fruit, and seed were analyzed through gas chromatography and mass spectroscopy and presented in (Fig. 1-4 and Table 1- 4).
Metabolite Profiling of Leaf Extracts: GC-MS is the best technique to identify the constituents of volatile matter, long-chain, branched-chain hydrocarbons, alcohols acids, esters, etc. Peak area, retention time, and the molecular formula used for the confirmation of phytochemical compounds. The active principles with their Retention time (RT), Molecular formula, Molecular weight (MW) and peak area in percentage presented. GC/MS analysis of an ethanolic extract of leaf, fruit and seeds of Syzyzium cumini revealed the existence of 105 compounds in the leaf extract. Of which carboxylic acid (19%), organic compound (11%), fatty acid (10%), carbohydrates (9%), sugars (7%), amino acid (6%), ester (6%), sugar alcohol (5%), alkane hydrocarbon (5%), heterocyclic compound (4%), phenols (3), hydrocarbon (3%), alcohol (2%), amine (2%), steroid (2%), alkaloid (2%), amine (1%), shikimic acid (1%) and amide (1%) found to be present in leaf extract metabolite profile. Whereas metabolite profiling of fruit extract revealed the presence of a total of 75 compounds. Of which fatty acids (11%), carbohydrates (8%), carboxylic acid (8%), organic acid (7%), amino acids (6%), sugars (5%), hydrocarbons (3%), heterocyclic compounds (3%), amides (3%), alkanes (3%), alkenes (3%), phenols (3%), sugar alcohol (3%), alcohol (2%), amine (2%), ester (2%), anilid (1%), ether (1%) and steroids (1%) were found to be present in fruit extract metabolite profile. And the metabolite profiling of seed extract revealed the presence of a total of 87 compounds. Of which carbohydrates (15%), fatty acids (13%), carboxylic acid (10%), dicarboxylic acid (9%), amino acids (8%), organic acids (6%), sugars (4%), alkane (3%), conjugated acid (3%), hydrocarbons (2%), organic compounds (2%), sugar alcohol (2%), heterocyclic compounds (2%), gondoic acid (1%), esters (1%), phenols (1%), amides (1%), amines (1%), phenacetin (1%), steroids (1%) and alkaloids (1%) were found to be present in seed extract metabolite profile.
TABLE 1 AND FIG. 1: THE ABUNDANCE OF EACH PRIMARY AND SECONDARY METABOLITES OF LEAF EXTRACTS OF SYZYGIUM CUMINI (L.)
TABLE 2 AND FIG. 2: THE ABUNDANCE OF EACH PRIMARY AND SECONDARY METABOLITES OF FRUIT EXTRACTS OF SYZYGIUM CUMINI (L.)
TABLE 3 AND FIG. 3: THE ABUNDANCE OF EACH PRIMARY AND SECONDARY METABOLITES OF SEED EXTRACTS OF SYZYGIUM CUMINI (L.)
FIG. 4: GC-MS ANALYSIS OF LEAF, FRUIT AND SEED EXTRACTS OF SYZYGIUM CUMINI L.
TABLE 4: APPLICATIONS OF METABOLITES OBSERVED IN LEAF, FRUIT, AND SEED OF S. CUMINI BY GC-MS ANALYSIS
|S. no.||Metabolite||Uses||Present in|
|1||6-Oxa-3-thiaoctanoic acid||Potential Inhibitor against Thyroid Cancer||Seed|
|2||Bisoprolol||Medicine for heart diseases specifically includes high blood pressure, chest pain from not enough blood flow to the heart, and heart failure.||Seed|
|3||Phenacetin||Phenacetin is a pain-relieving and fever-reducing drug||Seed|
|4||Tropine||Anticholinergic medicine is used to decrease the production of saliva and other fluid secretions during surgery. It is also used to treat spasms or cramps in the stomach, intestines, and other organs of the body||Seed|
|5||Nicotinaldehydethiosemicarbazone||Used in the treatment of pulmonary tuberculosis in adults||Leaf and seed|
|6||Alizarin / 1,2-Dihydroxyanthraquinone,||Used in carbon dating to identify carbonates||Seed|
|7||Pregnenolone||Pregnenolone is a precursor to many hormones. It relieves from blues or depression||Leaf, fruit, and seed|
|8||Gondoic acid/ 1,1-Eicosenoic acid,||Omega fatty acid||Seed|
|9||2-Butenedioic acid||Multiple sclerosis, psoriasis and to cure the autoimmune disorder||Leaf, fruit, and seed|
|10||Hexadecenoic acid||Antidiabetic, Anti-inflammatory, Antioxidant, hypocholesterolemia, nematicide, pesticide, and potent mosquito larvicide.||Leaf and seed|
|12||Octadecadienoic acid||Antidiabetic, Anti-cancer, Anti-inflammatory, antiandrogenic, cancer preventive, spermatogenic, hypo cholesterol emic and antidiabetic activity,||Seed and fruit|
|13||Tetra decanoic acid, trimethylsilyl ester||Antidiabetic, Larvicidal and repellent activity||Leaf|
|14||Benzoic acid trimethylsilyl ester||Intermediate in the biosynthesis of many secondary metabolites, anticancer and antimicrobial activity||Leaf, Fruit, and seed|
|15||Eicosane||Anti-androgenic and antimicrobial activity.||Fruit and seed|
|16||Heptadecanoic acid||antifungal and trypanocidal activity||Fruit and seed|
|17||Hexadecanoic acid||Antioxidant and antimicrobial activity||Fruit and seed|
|18||1,2,5,6-Hexanetetrol, tetrakis-O-(trimethylsilyl)-||Antifungal, antioxidant, antiallergic, antitumoral and antiparasitic activity||Fruit and leaf|
|19||Azelaic acid, bis(trimethylsilyl) ester||Acts as a "distress flare" involved in defense responses after infection. An important component of the plant's defensive response. Anti-inflammatory, anticancer and tyrosine inhibitor||Leaf|
|20||Penicillamine, tri-TMS||Treatment of Wilson's disease. Treatment for kidney stones which have high urine cystine levels, rheumatoid arthritis, and various heavy metal poisonings||Leaf|
|22||Octadecanoic acid||anti-inflammatory, antiviral and antiasthmatic||Leaf, fruit, and seed|
|23||Tetracosane||Antimicrobial, Antidiabetic, and antioxidant||Leaf, fruit, and seed|
|24||Tridecane, 5-propyl-||Antimicrobial agent||Leaf, fruit, and seed|
DISCUSSION AND CONCLUSION: 6-oxa-3-thiaoctanoic acid compound found in the seed is a potential inhibitor against thyroid cancer. Bisoprolol metabolites found in the seed used as medicine for heart diseases like high blood pressure, chest pain due to less blood flow to the heart and heart failure. Phenacetin compound found in the seed is a pain-relieving and fever-reducing drug. Tropine metabolite found in the seed is used as anticholinergic medicine to decrease the production of saliva and other fluid secretions during surgery. It also used to treat spasms or cramps in the stomach, intestine, and other organs of the body 20.
Nicotinaldehydethiosemicarbazone found in leaf and seed is used in the treatment of pulmonary tuberculosis in adults. Alizarin / 1, 2. Dihydroxy-anthraquinone found in the seed is used in carbon dating to identify carbonates. Pregnenolone compound found in leaf, fruit, and seed is a precursor of many hormones and also relieves depression. Gondoic acid metabolite found in the seed is an omega fatty acid. 2-butenedioc acid found in leaf, fruit and seed used for multiple sclerosis, psoriasis and to cure the autoimmune disorder. Hexadecenoic acid compound found in leaf and seed is used as antidiabetic, anti-inflammatory, antioxidant, nematicide and pesticide and potent mosquito larvicide 21.
Octadecadienoic acid found in seed and fruit used as antidiabetic, anti-inflammatory, antiandrogenic and cancer preventive. Tetra decanoic acid trimethylsilyl ester - found in the leaf used as an antidiabetic, anti larvicidal and anti-repellent activity. Benzoic acid trimethylsilyl ester metabolite found in leaf, fruit and seed is used for biosynthesis of many secondary metabolites, anticancer antimicrobial activity. Heptadecanoic acid compound found in fruit and seed is used as an antifungal and for trypanocide activity. Hexa-decanoic acid found in fruit and seed is used for antioxidant and antimicrobial activity.1,2,5,6-Hexanetetrol a compound found in fruit and leaf is used as an antifungal, anti-oxidant, anti-allergic, antitumoral and anti-parasitic activity. Azelaic acid bis (trimethylsilyl) ester found in leaf acts as a distress flare after infection, defensive response, anti-inflammatory, anti-cancer, and tyrosine inhibitor. Penicillamine, tri-TMS metabolite found in the leaf used to treat Wilson’s disease treatment for kidney stones and rheumatoid arthritis. S (-)-Cathinone found in leaf used as an antidepressant. Octadecanoic acid compound found in leaf, fruit, and seed used to cure asthma, anti-inflammatory, and antiviral. Tetracosane metabolite found in leaf, fruit and seed is used as anti-microbial, antidiabetic and antioxidant and tridecane, 5-propyl compound found in leaf, fruit and seed as an antimicrobial agent 22. GC-MS technique is one of the effective technique for the studies related to medicinal properties and antibacterial properties of herbal extracts 23. There are studies conducted to deal with the downstream processing of anthocyanins from black plum (S. cumini) to obtain anthocyanins in a purified form 24 this study includes adsorption employing six different adsorbents. Among these, Amberlite XAD7HP showed the highest adsorption capacity (1.07 mg/mL of adsorbent) and desorption ratio (87.62%). Characterization and evaluation of anthocyanin pigments from Syzygium cumini fruit peels for their antioxidant efficacy and stability 25. Furthermore identified and reported by high-performance liquid chromatography (HPLC) three anthocyanins compounds as glucosides are delphinidin, petunidin, and malvidin. It can be concluded that the compounds present in leaf, fruit and seeds of Syzygium cumini is extensively used traditional ailment for the treatment of various diseases especially diabetes and their associated complications 26. The plant has many important compounds that confer the main characteristics of the plant. Further investigations needed with HPLC data to support with specifically identifiable fractions of compounds, with the intention that this plant could be exploited as an antioxidant agent, natural medicine, or a nutritional supplement. It also makes a potential medicinal plant and the plant parts studied can be used as a traditional medicine to cure various ailments.
ACKNOWLEDGEMENT: Dr. Jyothi Chaitanya Pagadala (JCP) is grateful to the Dr. D. S. Kothari Postdoctoral Fellowship (F.4-2/2006/ (BSR)/BL/ 15-16/0320), UGC for providing Dr.D.S Kothari Postdoctoral Fellowship to carry out this work. Authors are also highly grateful to Prof. G. Padmaja and Head Department of Plant Sciences, University of Hyderabad, for provided that facilities complete the GC-MS analysis and for constant support for the completion of this work. We thank Dr. Allam Vijaya Bhaskar Reddy, Assistant Professor, Taxonomist, Department of Botany, Nizam College, Osmania University, Hyderabad, for plant Authentication.
CONFLICTS OF INTEREST: Nil
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How to cite this article:
Chilumula S, Pagadala JC, Bramhanapalli M and Nelavelli LB: GC-MS metabolite profiling of bioactive compounds in leaf, fruit and seed extracts of Syzyzium cumini (L.). Int J Pharm Sci & Res 2021; 12(7): 3814-20. doi: 10.13040/IJPSR.0975-8232.12(7).3814-20.
All © 2013 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
S. Chilumula *, J. C. Pagadala, M. Bramhanapalli and L. B. Nelavelli
Department of Botany, University College of Science, Saifabad, Osmania University, Hyderabad, Telangana, India.
30 June 2020
31 December 2020
06 May 2021
01 July 2021