Identification of Bioactive Compounds Present in Kulthi (Macrotyloma uniflorum) Seed Extract by Gas Chromatography-Mass Spectrometry

 

Basheer Mohammed Abdullah*, Abdul Jaleel A.H,

Shaikh Ifrah Fatema, Jiyaullakhan Maulakhan Pathan

Department of Physics, Maulana Azad College,

Dr. Babasaheb Ambedkar Marathwada University, Aurangabad, India.

*Corresponding Author E-mail: bmasalhalmi2016@gmail.com

 

ABSTRACT:

The objectives of this research were to determine the chemical composition of the extract of Macrotyloma uniflorum seeds by using Gas chromatography–Mass Spectrometry analysis (GC-MS). GC-MS  is a matchless method to study and measure the quantity of organic volatile and semi-volatile compounds. Gas chromatography is employed to separates mixtures into individual components employing a temperature-controlled capillary column. Mass spectrometry is utilized to recognize a variety of components from their mass spectra. In the present study, volatile/semivolatile compounds present inM. uniflorum seed extract were analyzed. M. uniflorum seed extract isextracted by the soxhlet extraction method and then analyzed by GC-MS. A total of 28 compoundswere found and quantified in this study. The major bioactive compounds in M. uniflorum seed extract are: 1,7-Octadiyne, 1-Propanone, 1-phenyl-3-[2-( phenylmethoxy)phenyl] -(16.93%), Benzene, 1-methyl-4-(1,2,2-trimethylcyclopentyl)-, (R)-, Benzene, 1-(1,5-dimethyl-4-hexenyl)- 4-methyl- (13.88%) and à-Farnesene, 1,3,6,10-Dodecatetraene, 3,7,11-trimethyl- (Z,E)- (12.68%). The present investigation revealed that M. uniflorum was composed of variety of metabolites and therapeutic active substances as well as novel substances. These substances could be isolated and further empirically evaluated to confirm their biologic and medicinal activities as well as investigate their mechanism of action.

 

KEYWORDS: GC-MS, Macrotyloma uniflorum, Bioactive Compounds Extract.

 

 

INTRODUCTION:

Macrotyloma uniflorumbelongsto the family Fabaceae is commonly known as Kulthi in Hindi and horse gram in English. It is aherbaceous plant with annual branches, suberect or twining, leaflets 2.5-5 cm, and found extensively throughout Asia, Australia, Burma, Malaysia, Mauritius, and Africa. This plant is grown in most arid regions in the southern regions of India1,2.The seed length of this plant is from 6 to 8 mm and the width ranges from 3 to 4 mm. The seeds are a trapezoidal or somewhat rounded rectangle in pale shape to, orange-brown, dark reddish-brown, or completely black3. Many medicinal plants are used by Africans and Asians as fixed medications for their primary health care4-6.

 

The M. uniflorum (Fabaceae) is normally used to feed horses, though it is also commonly used in dishes. In traditional ayurvedic cuisine, M. uniflorum (Fabaceae) is considered a food with medicinal qualities. This crop is known in the southern regions of India as the pulse crop of the poor and is a powerful source of food grains rich in antioxidants and protein7,8. This plant is known for its very high resistance to disease from pathogenic bacteria. There are only a few fungal diseases like root rot and viral infections like a yellow mosaic disease. This plant has very strong resistance against attack by microbial flora2

 

Gas chromatography is employed to separates mixtures into individual components employing a temperature-controlled capillary column. Mass spectrometry is utilized to recognize a variety of components from their mass spectra.

 

Therefore, the present research is conducted to investigate the phytochemical constituents of M. uniform (Fabaceae) seeds using the GC-MS technique.

 

MATERIALS AND METHODS:

Collection of plant materials:

The M. uniflorum (kulthi) seeds were purchased from Aurangabad–Maharashtra- India.The selected seeds were identified and authenticated in the Department of Botany, Maulana Azad College,Aurangabad (MS).

 

Preparation of Seed extract:

The seeds M. uniflorum (kulthi) were selected and cleaned with great care and ground to powder using an electrical grinder. The required quantity of the powder of M. uniflorum was weighed and transferred tothe soxhlet apparatus, extraction was carried out at 50-55 °C, and amixture of methanol and chloroform (Chemifs Chemical Private Limited) (1:1) was used to extract substances from the powder. The process of extracting the substance from the powder continued for six hours. Then, the extract was filtered by using Whatman No.1, and the filtrate was dried by a water bath9,10. Kulthi seed extract was used as a sample for GC-Ms analysis.

 

GC-MS analysis of M. uniflorum extract:

GC-MS analysis of extract M. uniflorum seed swas performed using Thermo Scientific Triple Quadrupole GC-MS (Trace 1300 GC, Tsq 8000 triple quadrupole MS) equipped with TG5MS(30 × 0.25 μm ID × 0.25 μmdf). For GC-MS detection, Helium gas was used as a carrier gas at a constant flow rate of 1 ml/min, and an injection volume of 1 μl was employed. The injector temperature was maintained at 250 °C, the ion-source temperature was 250 °C, and the oven temperature was programmed from 230 °C. The oven temperature was maintained at 50 °C isothermal at 280 °C, Mass Spectra transfer line temperature. The compounds were detected in the range of 40- 450 amu by matching with NIST library11,12.

 

RESULTS AND DISCUSSION:

GC-MS chromatogram analysis of the extract of M. uniflorum seeds figure (1) showed Twenty-eight peaks which indicates the presence of Twenty-eight phytochemical constituents. The chemical compounds, molecular formula, and molecular weight are as shown in table (1). The major phytochemical constituents are : 1,7-Octadiyne , 1-Propanone, 1-phenyl-3-[2-(phenylmethoxy)phenyl]- at RT (8.77), Benzene, 1-methyl-4-(1,2,2 trimethylcyclopentyl)-, (R)-, Benzene, 1-(1,5-dimethyl-4-hexenyl)- 4-methyl- at RT (8.64), and à-Farnesene, 1,3,6,10-Dodecatetraene, 3,7,11-trimethyl-, (Z,E)- at RT (8.84).

 

The extract of M. uniflorum seeds contains compounds with biological activities such asTrans-13-Octadecenoic acid act as an acidifier, arachidonic acid inhibitor, increase aromatic amino acid decarboxylase activity, inhibit the production of uric acid, urine acidifier, anti-inflammatory, antiandrogenic, cancer preventive, dermatitigenic, irritant, anti leukotriene—D4, hypocholesterolemic, 5-alpha reductase inhibitor, anemiagenic, insectifuge and flavor 13,14, cis-13-Octadecenoic acid act as anti-inflammatory, antiandrogenic, anticancer, preservative and hypocholesterolemic15,16, n-Hexadecanoicacidis reported to be an antioxidant, HypocholesterolemicNematicide, Pesticide, Lubricant, Antiandrogenic, Flavor, Hemolytic 5-Alpha reductase inhibitor17,18, 9,12-Octadecadienoic acid (Z,Z)-.‏ act as Antiinflammatory, Nematicide, Insectifuge, Hypocholesterolemic, Cancer preventive, Hepatoprotective, Antihistaminic, Antiacne, Antiarthritic, Antieczemic16, Hexadecanoic acid, methyl ester act as Antioxidant, Flavor, Hypocholesterolemic Pesticide, 5-Alpha reductase inhibitor19, 10-Octa decenoic acid, methyl ester act as Antioxidant and antimicrobial activities20. 1,6,10-Dodecatrien-3-ol, 3,7,11-trimethyl- act as Antitumor, analgesic, antibacterial, anti-inflammatory, sedative, fungicide21., Thymol act as antibacterial, antifungal, insecticidal, and anti-oxidative properties22., L-à-Terpineol act as Anticonvulsant, This compound is widely used in the perfumery, cosmetic, and soap industries. It is also used as a scenting agent in household products (e.g., disinfectant sprays)23.,1,7,7-Trimethylbicyclo[ 2.2.1]heptan-2-ol acts as an antiviral, antimicrobial, antitussive, and analgesic agent and is a natural product used since antiquity in awide range of applications, such as in food flavourings, fumigants, perfumes, cosmetics, household cleaners, and topically applied analgesics in the medical chemistry field24.

 

Thus, GC- MS analysis of M. uniflorum seed extract is the first step towards understanding the active compounds in this plant, and sucha study would be helpful for a more detailed study. Further investigation into the pharmacological importance of M. uniflorum and their diversity and detailed phytochemistry may add new knowledge to the information in the traditional medicinal plants.

 

Table 1: Compounds with Reported Biological Activities present in kulthi (M. uniflorum) seed extract:

Sr. No

Compound Name

Area %

Molecular formula

Molecular Weight

RT (min)

Reported Bioactivity

Structure

1

cis-13-Octadecenoic acid

10.20

C18H34O2

 

282

17.38

anti-inflammatory, antiandrogenic, anticancer, preservative, and hypocholesterolemic

 

2

Trans-13-Octadecenoic acid

0.79

C18H34O2

282

14.40

acidifier, arachidonic acid inhibitor, increase aromatic amino acid decarboxylase activity, inhibit the production of uric acid, urine acidifier, anti-inflammatory, antiandrogenic, cancer preventive, dermatitigenic, irritant, anti leukotriene—D4, hypocholesterolemic, 5-alpha reductase inhibitor, anemiagenic, insectifuge and flavor

 

3

9,12-Octadecadienoic acid (Z,Z)-.‏

14.33

C18H32O2

280

1.60

Antiinflammatory, Nematicide, Insectifuge, Hypocholesterolemic, Cancer preventive, Hepatoprotective, Antihistaminic, Antiacne, Antiarthritic, Antieczemic,

 

4

Hexadecanoic acid, methyl ester

 

1.36

C17H34O2

270

11.89

Antioxidant, Flavor, Hypocholesterolemic Pesticide, 5-Alpha reductase inhibitor

 

 

5

n-Hexadecanoic acid

 

1.79

C16H32O2

256

12.32

antioxidant, HypocholesterolemicNematicide, Pesticide, Lubricant, Antiandrogenic, Flavor, Hemolytic 5-Alpha reductase inhibitor

 

 

6

10-Octa decenoic acid, methyl ester

 

0.46

C19H36O2

296

13.67

Antioxidant and antimicrobial activities.

 

 

7

1,6,10-Dodecatrien-3-ol, 3,7,11-trimethyl-

 

0.96

C15H26O

222

10.31

Antitumor, analgesic, antibacterial, anti-inflammatory, sedative, fungicide

 

 

8

Thymol

 

1.03

C10H14O

150

6.98

antibacterial, antifungal, insecticidal, and anti-oxidative properties

 

9

L-à-Terpineol

 

0.42

C10H18O

154

5.94

Anticonvulsant,This compound is widely used in the perfumery, cosmetic, and soap industries.

 

10

1,7,7-Trimethylbicyclo[

2.2.1]heptan-2-ol

0.59

C10H18O

154

5.68

antiviral, antimicrobial, antitussive, and analgesic agent

 

 

 

 

Figure.1: GC-MS Chromatogram of Macrotylomaunifloruextract.

 

Table 2: Probable Compounds Present in M. uniflorum extract.

S No.

RT

Area %

Compound Name

Molecular Formula

Molecular weight

1

5.68

0.59

Endo-Borneol

C10H18O

154

1,7,7-Trimethylbicyclo[2.2.1]heptan-2-ol

C10H18O

154

Bicyclo[2.2.1]heptan-2-ol,1,7,7-trimethyl-,(1Sendo)-

C10H18O

154

2

5.94

0.42

L-à-Terpineol

C10H18O

154

À-Terpineol

C10H18O

154

3-Cyclohexene-1-methanol,à,à,4-trimethyl-,acetate

C12H20O2

196

3

6.98

1.03

Thymol

C10H14O

150

Phenol, 2-methyl-5-(1-methylethyl)-

C10H14O

150

3-Methyl-4-isopropylphenol

C10H14O

150

4

8.45

0.42

Spiro[5.5]undec-2-ene,3,7,7-trimethyl-11-methylene-,(-)-

C15H24

204

1H-Benzocycloheptene,2,4a,5,6,7,8,9,9a-octahydro-3,5,5-trimethyl-9-methYlene-,(4aS-cis)-

C15H24

204

5

8.64

13.88

Benzene,1-methyl-4-(1,2,2-trimethylcyclopentyl)-

,(R)-

C15H22

202

Benzene, 1-(1,5-dimethyl-4-hexenyl)-4-methyl-

C15H22

202

6

8.77

16.93

1,7-Octadiyne

C8H10

106

1-Propanone,1-phenyl-3-[2-(phenylmethoxy)phenyl]-

C22H20O2

316

7

8.84

12.68

À-Farnesene

C15H24

204

1,3,6,10-Dodecatetraene,3,7,11-trimethyl-,(Z,E)-

C15H24

204

Bicyclo[3.1.1]hept-2-ene,2,6-dimethyl-6-(4-methyl-3-pentenyl)-

C15H24

204

8

8.99

10.48

1H3a,7Methanoazulene,octahydro3,8,8trimethyl6methylene,

[3R(3à,3aá,7á,8aà)]

C15H24

204

Cyclohexene,3(1,5dimethyl4hexenyl)6methylene,[S( R*,S*)]

C15H24

204

Cedrene

C15H24

204

9

9.14

0.37

áGuaiene

C15H24

204

10

9.26

1.37

1,6,10-Dodecatrien-3-ol,3,7,11-trimethyl-,(E)-

C15H26O

222

1,6,10-Dodecatrien-3-ol,3,7,11-trimethyl-

C15H26O

222

1,6,10-Dodecatrien-3-ol, 3,7,11-trimethyl-, [S-( Z)]-

C15H26O

222

13

9.99

2.48

8-(2-Acetyloxiran-2-yl)-6,6-dimethylocta-3,4-dien-2-one

C14H20O3

236

14

10.31

0.96

6,10-Dodecadien-1-yn-3-ol,3,7,11-trimethyl-

C15H24O

204

15

11.15

3.15

Spiro[4.5]dec-6-en-8-one,1,7-dimethyl-4-(1-methylethyl)-

C15H24O

204

16

11.89

1.36

Hexadecanoic acid, methyl ester

C17H34O2

270

Pentadecanoic acid, 14-methyl-,methyl ester

C17H34O2

270

Hexadecanoic acid, 15-methyl-,methyl ester

C18H36O2

284

17

12.32

1.79

n-Hexadecanoicacid

C16H32O2

256

l-(+)-Ascorbicacid 2,6-dihexadecanoate

C38H68O8

652

Pentadecanoic acid

C15H30O2

242

18

13.60

1.50

9,12-Octadecadienoicacid (Z,Z)-,methyl ester

C19H34O2

294

8,11-Octadecadienoicacid, methyl ester

C19H34O2

294

19

13.67

0.46

10-Octadecenoicacid, methyl ester

C19H36O2

296

16-Octadecenoicacid, methyl ester

C19H36O2

296

20

13.92

1.55

Furan, 2,5-dibutyl-

C12H20O

180

Phenol, 2-methoxy-4-propyl-

C10H14O2

166

21

14.33

1.60

9,12-Octadecadienoicacid (Z,Z)-

C18H32O2

280

Oxacycloheptadec-8-en-2-one,(8Z)

C16H28O2

252

22

14.40

0.79

Trans-13-OctadecenoicAcid

C18H34O2

282

16-Octadecenoicacid, methyl ester

C19H36O2

296

Cis-13-OctadecenoicAcid

C18H34O2

282

23

17.38

10.20

p-Benzoquinone,

C22H36O2

332

24

18.26

9.52

Gingerol

C17H26O4

294

Acetamide,N-[2-[4-(acetyloxy)-3-methoxyphenyl]ethyl]-

C13H17NO4

251

25

18.93

0.78

Gingerol

C17H26O4

294

26

20.88

2.11

2-Cyclohexyl-2,5-cyclohexadiene-1,4-dione,4-oxime

C12H15NO2

205

28

21.80

0.62

1-[2-[2-Bromoacetamido]ethyl]hypoxanthine

C9H10BrN5O2

300

 

CONCLUSION:

The present investigation revealed that M. uniflorum extract was composed of a variety of metabolites and therapeutic active substances. These substances could be isolated and further empirically evaluated to confirm their biologic and medicinal activities as well as investigate their mechanism of action.

 

CONFLICT OF INTEREST:

The authors declare no conflicts of interest.

 

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Received on 10.11.2020            Modified on 24.04.2021

Accepted on 02.07.2021           © RJPT All right reserved

Research J. Pharm.and Tech 2022; 15(2):814-818.

DOI: 10.52711/0974-360X.2022.00135