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Research Article - International Research Journal of Plant Science ( 2022) Volume 13, Issue 2

Phytochemical analysis of Momordica dioica, a selected Indian medicinal plant by HR-LCMS spectra method

Swati Rangrao Jadhav and Laxmikant Haribhau Kamble*
 
School of Life Science, Swami Ramanand Teerth Marathwada University, Nanded-431 606, Maharashtra, India
 
*Corresponding Author:
Laxmikant Haribhau Kamble, School of Life Science, Swami Ramanand Teerth Marathwada University, India, Email: lhkamble@srtmun.ac.in

Received: 21-Mar-2022, Manuscript No. IRJPS-22-57894; Editor assigned: 22-Mar-2022, Pre QC No. IRJPS-22-57894(PQ); Reviewed: 05-Apr-2022, QC No. IRJPS-22-57894; Revised: 09-Apr-2022, Manuscript No. IRJPS-22-57894(R); Published: 16-Apr-2022, DOI: http:/dx.doi.org/10.14303/irjps.2022.007

Abstract

Plants have been used for various medical applications since the beginning of human history and are considered as the basis for modern medicines. Phytochemicals present in plants have already been reported as potential candidates in this regard. Due to the tremendous applications of medicinal plant products in the pharmaceuticals and biotechnology field, phytochemical analysis of medicinal plants has become an important and challenging task. An analytical technique like high resolution liquid chromatography mass spectrometry (HR-LCMS) is found to be an important technique in the analysis of complex bioactive phytoconstituents. The present study was aimed at bioactive constituent analysis from Momordica dioica fruits by using HR-LCMS analysis. The study confirms presence of compounds having potential of being therapeutic agents, which includes alkaloid, flavonoid, phenol, saponins, cardiac glycosides, tannin, carbohydrates, terpenoids and steroids.

Keywords

HR-LCMS, Momordica dioica, Phytoconstituents

Introduction

The role of phytoconstitutes extracted from medicinal plants in maintaining sustainable human health documented worldwide. The traditional medicinal practices including Ayurveda, Rig-Veda (3700 B.C.), Unani and Homeopathy mentioned the use of medicinal plant products for the cure of various human diseases (Balkrishna et al., 2017; Pandey, 2013). In the last few years, many drugs were explored with low side effects from medicinal plants. There is an increasing demand for the identification of novel, potent drug molecules from medicinal plant products that are safe with low side effects to treat various diseases (Lahlou, 2013; Patra, 2012). In the phytochemical analysis of plant, the first step is the identification and isolation of bioactive phytoconstitute from the medicinal plants.

It is well known that some plant products and vegetables, which were used as dietary supplements, might reduce the effects of cancer proliferation. Hence, ethno-medicinal plants had tremendous contribution in the development drugs to prevent or treat various diseases, including the cancer also (Fatma et al., 2019). Their preventive effects might induce a decrease in cell proliferation as well as reduce cancer invasion and spread. It has been proposed that the whole-plant effects might be much better than its active components (Aggarwal et al., 2013).

In the present study, Momordica dioica plant was selected from the native places of Nanded district and their extracts were analyzed using HR-LCMS for the identification of bioactive molecules. The plant was traditionally used as an astringent, febrifuge, antiseptic, antihelmintic, antibacterial, anti-inflammatory, hepatoprotective, hypoglycemic and analgesic properties (Bawara et al., 2010). The fruits of Momordica dioica shows various medicinal properties like analgesics, anti-tumorogenic, anti-inflammatory, antidiabetic activity and anti-cancer activity (Ahirrao et al., 2019).

Materials and Methods

Collection of sample

The fruits of the Momordica dioica plant were collected from the rural areas of Nanded district. The fruits were cleaned by washing thoroughly 2-3 times with running tap water and once sterile distilled water. It was followed by cutting into small pieces, shade drying, grinding and storing in well closed containers for further use (Revathy et al., 2015).

Extraction of Bioactive Compounds

The fruits of Momordica dioica were finely powdered and bioactive compounds were extracted with petroleum ether and acetone using a Soxhlet extractor (Redfern et al., 2014). The extracts were then collected and stored at 4°C for further analysis.

High Resolution-Liquid Chromatography Mass Spectrometry (HR-LCMS) Methodology

The fruit extracts of Momordica dioica prepared in petroleum ether and acetone were subjected to HRLCMS analysis individually and chemical fingerprints were prepared using high-resolution liquid chromatography and mass spectrometry (model-G6550A of Agilent technologies) with 0.01% mass resolution (Pitt, 2009) with following parameters:

a. MS- minimum range 150 (M/Z) and maximum 1000 daltons with scanning rate each per second.

b. The source parameter for gas chromatography was maintained at 250°C with a gas flow of 13 psi/minute.

c. The auxiliary draw speed was 100 μl/minute, eject speed at 100.0 μL/min, draw position offset 0.0 mm wait time after drawing 2.0 s, Sample flush out factor was 5.0 (Tables 1 and 2).

Table 1. Solvent Composition.

Sl. no Channel Ch. 1 Solv. Name 1 Ch2 Solv. Selected Used Percent
1 A 100.0% Water
V.02
0.1% FA in
water
100.0% Water
V.02
Ch. 1 Yes 95.00%
2 B 100.0%
Acetonitrile
V.02
90% ACN +10%
H2O+ 0.1% FA
100.0%
Acetonitrile
V.02
Ch. 1 Yes 5.00%

Table 2. Timetable.

Sl. no Time A B Flow Pressure
1 1.00 min 95.00% 5.00% 0.300 mL/min 1200.00 bar
2 20.00 min 0.00% 100.00% 0.300 mL/min 1200.00 bar
3 25.00 min 0.00% 100.00% 0.300 mL/min 1200.00 bar
4 26.00 min 95.00% 5.00% 0.300 mL/min 1200.00 bar
5 30.00 min 95.00% 5.00% 0.300 mL/min 1200.00 bar

Results and Discussion

The High Resolution-Liquid Chromatography-Mass spectrometry analysis (HR)-LCMS of petroleum ether extract of Momordica dioica fruit was found to contain 38 compounds which were confirmed based on their mass and molecular formula as shown in Table 3, chromatogram Figure 1. The chromatogram gives information on the relative concentrations of various compounds eluted as a function of retention time.

Table 3. Bioactive Compounds in petroleum ether extract of Momordica dioica fruit.

Sl. No. Name of compound Compound formula Mass
1 Clenbuterol C12 H18 Cl2 N2 O 276.0816
2 Lycoperdic acid C8 H11 N O6 217.0619
3 Thiabendazole C10 H7 N3 S 201.0325
4 3-tert-Butyl-5-methylcatechol C11 H16 O2 180.1146
5 19-Noretiocholanolone C18 H28 O2 276.2081
6 Beta-Cortol C21 H36 O5 368.2556
7 1-Naphthylacetylspermine C22 H34 N4 O 370.2713
8 Triphenyl phosphate C18 H15 O4 P 326.07
9 9Z-Octadecen-12-ynoic acid C18 H30 O2 278.2241
10 Linoleoyl Ethanolamide C20 H37 N O2 323.2818
11 3-Methylcyclopentadecanone C16 H30 O 238.2319
12 Camelledionol C29 H44 O3 440.3296
13 Palmitic amide C16 H33 N O 255.256
14 Oleamide C18 H35 N O 281.2716
15 Monoolein C21 H40 O4 356.2918
16 4'-Apo-beta,psi-caroten-4'-carotenal C35 H46 O 482.3593
17 Corchorifatty acid F C18 H32 O5 328.2297
18 9Z-Octadecenedioic acid C18 H32 O4 312.2352
19 Dibutyl decanedioate C18 H34 O4 314.2508
20 Estradiol-17-phenylpropionate C27 H32 O3 404.2379
21 Sorbitan laurate C18 H34 O6 346.2328
22 Nandrolone phenpropionate C27 H34 O3 406.2535
23 12S,13R-EpOME C18 H32 O3 296.2405
24 Milbemectin C31 H44 O7 528.315
25 Practolol C14 H22 N2 O3 266.1596
26 Lauryl hydrogen sulfate C12 H26 O4 S 266.16
27 α-Linolenic Acid C18 H30 O2 278.2296
28 Linalyl caprylate C18 H32 O2 280.2457
29 Docosanedioic acid C22 H42 O4 370.3161
30 Carpaine C28 H50 N2 O4 478.3768
31 Isopalmitic acid C16 H32 O2 256.2453
32 Praziquantel C19 H24 N2 O2 312.1819
33 Pachymic acid C33 H52 O5 528.3936
34 Petroselinic acid C18 H34 O2 282.2618
35 Rhodoxanthin C40 H50 O2 562.3744
36 N-Nitrosotomatidine C27 H44 N2 O3 444.3351
37 Stearic acid C18 H36 O2 284.2775
38 Calpeptin C20 H30 N2 O4 362.2211
Figure

Figure 1. HR-LCMS Spectrogram of petroleum ether extracts of Momordica dioica fruit.

The height of the peak indicates the relative concentrations of bioactive compounds. Mass Spectrometer analyses the structure of unknown compounds which are eluted at different times. The important phytoconstituents confirmed by HR-LCMS Analysis were Clenbuterol, Lycoperdic acid, Palmitic amide, Oleamide, Milbemectin, etc. The compounds have reported various activities like antioxidant, antineoplastic, antiviral, anticarcinogenic, antiviral. Most of them were prominently reported anticancer activity. Lycoperdic acid shows the anticancer activity in the form of dietary phenolics compound which is used in cancer treatment (Anantharaju et al., 2016). The Palmitic amides were reported in the treatment for bladder cancer in the form of heterocyclic derivative of fatty acids (Jozwiak et al., 2020). Oleamide has shown the anticancer activity against the MDA-MB-231 Cell Line in In vitro Bioassay (Wisitpongpun et al., 2020). Milbemectin has shown the anticancer activity against leukemia (El-Saber et al., 2020) (Figure 1).

The Phytochemicals found in the extract including Adenosine, Cucurbic acid, Leukotriene E3, Methanophenazine, Momordicoside I, Vulgarone A, Pyropheophorbide a, Camelledionol, Azelaic acid, Retamine, Petroselinic acid were shown in Table 4. It was also reported that these compounds found in the different species of plants exhibit different pharmacological activities (Tsuchiya & Nishizaki, 2015). Among these: Leukotrienes are lipid mediators which play impotant roles in acute and chronic inflammation and allergic diseases. They also play roles in various allergic diseases, including asthma, atopic dermatitis, allergic rhinitis, allergic conjunctivitis and anaphylaxis (Jo-Watanabe et al., 2019). Pyropheophorbide a isolated from G. elliptica is a potential glioblastoma-specific anticancer agent without side effects on normal cells. In addition, specifically it had cytostatic activity on glioblastoma cells rather than human umbilical vein endothelial cells (Cho et al., 2014). The in vitro cytotoxic activity of azelaic acid was studied with 25 human melanoma primary cultures and with 5 established cell lines characterized by different contents of melanotic pigment (Zaffaroni et al., 1990).

Table 4. Bioactive Compounds in acetone extract of Momordica dioica fruit.

Sl. No. Name of compound Compound formula Mass
1 Adenosine C10 H13 N5 O4 267.0967
2 Butopyronoxyl C12 H18 O4 226.1201
4 Isocarbostyril C9 H7 N O 145.0523
5 Cucurbic acid C12 H20 O3 212.1403
6 Leukotriene E3 C23 H39 N O5 S 441.2493
7 Dasytrichone C18 H16 O4 296.1045
8 Dihydrodeoxystreptomycin C21 H41 N7 O11 567.2884
9 3-tert-Butyl-5-methylcatechol C11 H16 O2 180.1147
10 Aegle marmelos Alkaloid C C23 H27 N O3 365.1969
11 9Z-Octadecen-12-ynoic acid C18 H30 O2 278.2241
12 Methanophenazine C37 H50 N2 O 538.3879
13 Momordicoside I C36 H58 O8 618.4123
14 Vulgarone A C15 H22 O 218.1667
15 LysoPE(24:0/0:0) C29 H60 N O7 P 565.4196
16 Islanditoxin C24 H31 Cl2 N5 O7 571.1714
17 Linoleoyl Ethanolamide C20 H37 N O2 323.2819
18 3-Ketosphinganine C18 H37 N O2 299.2843
19 Epoxyganoderiol C C30 H48 O3 456.3593
20 Oleoyl Ethanolamide C20 H39 N O2 325.2974
21 4,4'-Methylenebis(2,6-di-tert-butylphenol) C29 H44 O2 424.3342
22 Pheophorbide a C35 H36 N4 O5 592.2677
23 Pyropheophorbide a C33 H34 N4 O3 534.2621
24 Camelledionol C29 H44 O3 440.3314
25 4'-Apo-beta,psi-caroten-4'-al C35 H46 O 482.3597
26 Azelaic acid C9 H16 O4 188.1071
27 Hericenone B C27 H31 N O4 433.2289
28 Cilazapril C22 H31 N3 O5 417.2344
29 Muricatacin C17 H32 O3 284.2412
30 2alpha-Fluoro-17beta-hydroxyandrost-4-en-3-one C19 H27 F O2 306.2009
31 Corchorifatty acid F C18 H32 O5 328.231
32 Phygrine C16 H28 N2 O2 280.2096
33 Pimozide C28 H29 F2 N3 O 461.2252
34 Retamine C15 H26 N2 O 250.1993
35 Momordin Ia C42 H66 O13 778.4667
36 Sorbitan laurate C18 H34 O6 346.2333
37 Phlegmarine C16 H30 N2 250.2357
38 Formimidoyl-fortimicin A C18 H36 N6 O6 432.2736
39 (-)-Ormosanine C20 H35 N3 317.2794
40 Practolol C14 H22 N2 O3 266.1602
41 Ricinoleic acid C18 H34 O3 298.2572
42 Ethyl 2E,4Z-hexadecadienoate C18 H32 O2 280.2465
43 Petroselinic acid C18 H34 O2 282.2627
44 Homodolicholide C29 H48 O6 492.3574
45 2-Dodecylbenzenesulfonic acid C18 H30 O3 S 326.1956

The HR-LCMS High analysis of acetone extract of Momordica dioica fruit spectrum profile (Figure 2) shows 45 compounds which were confirmed based on their retention time, mass and molecular formula.

Figure

Figure 2. HR-LCMS Spectrogram of acetone extracts of Momordica dioica fruit.

Conclusion

The petroleum ether and acetone extract of Momordica dioica fruits revealed the presence of therapeutically important bioactive phytocompounds like alkaloids, Flavonoids, Phenols, Saponins, Cardiac glycosides, Tannins, Carbohydrates, Terpenoids and Steroids using (HR)-LCMS high-resolution liquid chromatography-mass spectrometer analysis. These bioactive phytoconstitutes possess important pharmacological activities and could be useful for treating various human ailments.

Acknowledgement

The authors are immensely thankful to the DST-FIST and UGC SAP-DRS-Phase-II sponsored school of life science, Swami Ramanand Teerth Marathwada University, Nanded for providing the infrastructure and necessary facilities. Sophisticated Analytical Instrument Facility (SAIF), IIT Bombay for HR-LCMS spectroscopy support. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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Citation: Jadhav SR & Kamble LH (2022). Phytochemical analysis of Momordica dioica, a selected Indian medicinal plant by HR-LCMS spectra method. IRJPS.13: 007.