Journal of Physics: Conference Series

PAPER • OPEN ACCESS

Recent citations
Gas Chromatographic Study of Bio-active - Agro-waste derived compounds (flax and
Compounds in Methanolic Extract of Leaf of black seed peels): Toxicological effect
against the West Nile virus vector, Culex
pipiens L. with special reference to
Crateva adansonii DC GC–MS analysis
Shaimaa M. Farag et al

To cite this article: O. Ajanaku Christiana et al 2019 J. Phys.: Conf. Ser. 1299 012014

View the article online for updates and enhancements.

This content was downloaded from IP address 165.73.223.225 on 27/09/2021 at 18:31

3rd International Conference on Science and Sustainable Development (ICSSD 2019) IOP Publishing
IOP Conf. Series: Journal of Physics: Conf. Series 1299 (2019) 012014 doi:10.1088/1742-6596/1299/1/012014

Gas Chromatographic Study of Bio-active Compounds in

MethanolicExtract of Leaf of Crateva adansonii DC

Ajanaku Christiana O.1, EchemeJohnbull O.2, Mordi Raphael C.1, Olugbuyiro

Joseph O.1, Osamudiamen Paul M.3, Jolayemi Emmanuel G.1
1
Department of Chemistry, P.M.B.1023, Ota, Ogun State, Nigeria.
2
Department of Chemistry, Michael Okpara University of Agriculture, Umudike,
Abia State, Nigeria.
3
Department of Chemical and Food Science, Bells University of Technology,
Ota.Ogun State.

Corresponding author’s email: [email protected]

Abstract. This study investigates the volatile bioactive components in Crateva adansonii plant
that has been applied in the treatment of diverse ailments in the Western part of Nigeria and in
Africa. The methanolic leaf extract of Crateva adansonii was subjected to analysis using gas
chromatographic-mass spectrometric technique. The mass spectra of the compounds were
analysed and identity of the compounds confirmed using the data base of the National
Institute of Standard and Technology (NIST) library. The compounds identified includesn-
Hexadecanoic acid; 9,12-Octadecadienoic acid (Z,Z); 9-Octadecenoic acid; (Z,E)-2,13-
Octadecadien-1-ol; 9,17-Octadecadienal; cis-9-Hexadecenal; cis-Vaccenic acid; Z,Z-10,12-
Hexadecadienal; 13-Octadecenal; Hexadecanoic acid 2-hydroxy-1-(hydroxymethyl)ethyl
ester; E-11-Hexadecenal. Most of the compounds identified have antibacterial, anti-oxidant,
anti-inflammatory and hypocholesterolemic properties, which affirms the therapeutic
applications of Crateva adansonii as a medicinal plant.
Keywords: Crateva adansonii, GC-MS, medicinal plants, ethnobotany, antimicrobial

INTRODUCTION
Medicinal plants have been important tools as food, for prevention and treatment of
ailments. Several modern medicines that have been in use in the treatment of various ailments
have their origin from plants. It is imperative, therefore, to identify the chemically active

Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution
of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI.
Published under licence by IOP Publishing Ltd 1
3rd International Conference on Science and Sustainable Development (ICSSD 2019) IOP Publishing
IOP Conf. Series: Journal of Physics: Conf. Series 1299 (2019) 012014 doi:10.1088/1742-6596/1299/1/012014

constituents of such medicinal plants. This will further advance the synthesis of drugs that are
therapeutic[1]. The application of medicinal plants as therapeutic agents began as far back as
2600 B.C. with the Egyptians, the Greeks, Chinese, and Romans which applied the use of
natural products in diverse forms ranging from decoctions, gargles, ointments, pills and
infusions being used to treat disorders ranging from mild to complex states [2].Crateva
adansonii, (CA) is a deciduous plant, found mainly in Africa. It grows to a height of about 10
metres. The plant, Crateva adansonii is from the family, Capparaceae and species C.
adansonii, commonly distributed across the savannah and forest areas across Nigeria [3]. It
has close similarity with another species, Crateva religiosa G. Forst from Asia, which is
reported to exhibit same properties [4].Thebole is short and irregular. The surface of its bark
is smooth, while the inner part of the bark is yellow. The leaves are in alternate positions, they
are clustered at the end of the twigs, with 3 leaflets.
The leaves are used in sauces, soups and prepared as cooked vegetables. Decoctions from
the leaf part of the plant is functional as a vapour bath against yellow fever, eye complaints
and jaundice [5]. It is also taken orally in the treatment of pains, malaria[6][7]
hypertension[8], oedema, jaundice, epilepsy, skin diseases [9]dysmenorrhoea and
abscesses[10]. The dried and ground leaves are used to stimulate production of milk for
lactating mothers in Burkina Faso [4] An earlier study on the methanolic extract of the leaf of
CA against free radicals and antibiotic resistant organisms revealed the anti-oxidant and
antibiotic potentials in the plant [7] [11]. The phytochemical constituents and antimicrobial
properties of this plant have been reported[12]. An important technological platform for
profiling of volatile secondary metabolites in plant species is the employment of gas
chromatography-mass spectrometry (GC-MS). Here we report on the gas chromatography
coupled with mass spectrometry study of the leaf extract of CA. This study aims to investigate
the volatile chemical components from the leaf of CA. It commences by first preparing the
methanolic extract of the leaf, separation and identification of its components by subjecting
the extract to GC-MS analysis.

MATERIALS AND METHODS

Collection of plant materials

2
3rd International Conference on Science and Sustainable Development (ICSSD 2019) IOP Publishing
IOP Conf. Series: Journal of Physics: Conf. Series 1299 (2019) 012014 doi:10.1088/1742-6596/1299/1/012014

The leaves of CA plant were collected from Iyesi village, in Ado-Odo/Ota local
government of Ogun State, Nigeria. The plant identification was carried out by a botanist at
the Forestry Herbarium, Forestry Reserve Institute of Nigeria, (FRIN), Ibadan, with No. FHI
110016.
Preparation of Plant Extracts
Dried leaves of CA were soaked in methanol (500 mL) and left for 72 hours. The mixture
was filtered and the filtrate concentrated on a rotary evaporator. The crude extract of C.
adansonii obtained was kept in the refrigerator for further analysis.

Gas Chromatography-Mass Spectrometry analysis

Analysis of the methanolic leaf extract of the CA plant was carried out on a GC-MS
equipment by Agilent 7890A. The experimental conditions of the equipment are: HP-5MS
ultra inert capillary non-polar column, dimensions: 30 mm × 0.25 mm; ID: 0.25 mm, film
thickness: 0.25 μm. Flow rate of mobile gas: 1.0 ml/min. The oven temperature for the gas
chromatographic part was 50°C raised to 300°C at 7oC/min for 10 min. The nature and
structure of compounds were identified by the mass spectrometer. Appearance of peaks on the
spectrum is as a result of the fragmentation of large compounds into smaller compounds at
various m/z ratios. Spectrum of unidentified components was compared with the spectrum of
identified components stored in the NIST library [13][14].

RESULTS AND DISCUSSION

The results of the study are presented in Table 1. One of the current techniques employed
in compound identification from natural products is the gas chromatography equipment. It
offers a simple and fast analytical approach to identification of low molecular weight
compounds in plant extracts. Volatile components of extracts can be easily identified using
this equipment. Compounds identified from the chromatographic analysis of the methanolic
extract of the leaf of C. adansonii include: n-Hexadecanoic acid; 9,12-Octadecadienoic acid
(Z,Z); 9-Octadecenoic acid; (Z,E)-2,13-Octadecadien-1-ol; 9,17-Octadecadienal; cis-9-
Hexadecenal; cis-Vaccenic acid; Z,Z-10,12-Hexadecadienal; 13-Octadecenal; Hexadecanoic
acid 2-hydroxy-1-(hydroxymethyl)ethyl ester; E-11-Hexadecenal. From table 1, the retention
time, peak area, molecular weight and the molecular formula constituted the parameters by
which each compound was identified. Compounds identified in this study show similarity

3
3rd International Conference on Science and Sustainable Development (ICSSD 2019) IOP Publishing
IOP Conf. Series: Journal of Physics: Conf. Series 1299 (2019) 012014 doi:10.1088/1742-6596/1299/1/012014

with earlier studies on the chromatographic analysis of C. religiosa[15]. From Table 1, the
compound identified with the least retention time is n-Hexadecanoic acid (25.61min) while E-
11-Hexadecenal, was the last compound identified with the longest retention time (40.30
min). Figure 1, shows the chromatogram of the methanolic extract of the leaf of CA. The
figure shows the various compounds identified alongside their retention times. Nature of
compounds identified from the table can be categorized as fatty acids, long chain unsaturated
aldehyde, fatty acid esters and polyunsaturated fatty acids (PUFA).
Fats are known to be vital sources of energy however, fats, as dietary intakes has more
roles to the physiological system. Earlier studies have shown that unsaturated fatty acids have
more health benefits than saturated fatty acids[16][17] and[18]. Observations made on intake
of dietary fats have shown a steady relationship between polyunsaturated fatty acids and
reduced risk of heart disease[19]. The compounds identified in Table 1 have common
bioactive properties which include: antioxidant, anti-inflammatory [20], hypocholesterolemic,
cancer preventive, antifungal, antibacterial, anti-acne, anti-coronary, anti-eczemic, insecticidal
properties[21]. Unsaturated fatty acids can be obtained from fish, vegetables, olive oil, cotton
seed oil, canola oil, or lean meat[22][23].
Table 1:Compounds identified in the methanolic leaf extract of Crateva adansonii in GC-MS
Ret Peak Compound Mol. Wt. Mol. formula Nature of
time area (g/mol) compound
(min) (%)
25.61 19.92 n-Hexadecanoic acid 256.42 C16H32O2 Fatty acid
(palmitic acid)
28.03 47.56 9,12-Octadecadienoic 280.45 C18H32O2 Polyunsaturated
acid (Z,Z) Fatty acid
29.45 0.31 9-Octadecenoic acid 282.10 C18H34O2 Fatty acid
29.53 0.28 (Z,E)-2,13- 266.469 C18H34O pheromone
Octadecadien-1-ol
31.99 0.91 9,17-Octadecadienal 264.453 C18H32O Long chain
unsaturatedaldeh
yde
32.81 2.03 cis-9-Hexadecenal 238.415 C16H30O Long chain
unsaturatedaldeh
yde
33.22 0.27 cis-Vaccenic acid 282.468 C18H34O2 trans-fatty acid
33.93 0.81 (Z,Z)-10,12- 236.399 C16H28O Long chain
Hexadecadienal unsaturatedaldeh
yde
36.12 0.86 13-Octadecenal 266.469 C18H34O Long chain

4
3rd International Conference on Science and Sustainable Development (ICSSD 2019) IOP Publishing
IOP Conf. Series: Journal of Physics: Conf. Series 1299 (2019) 012014 doi:10.1088/1742-6596/1299/1/012014

unsaturatedaldeh
yde
36.68 9.83 Hexadecanoic acid 2- 330.509 C19H38O4 Fatty acid ester
hydroxy-1-
(hydroxymethyl)ethyl
ester
40.30 0.56 E-11-Hexadecenal 238.415 C16H30O Long chain
unsaturatedaldeh
yde

Fig. 1: Chromatogram of methanolic leaf extract of Crateva adansonii

CONCLUSION
The search for lead compounds as a cure to numerous microbial issues from natural source
is endless, so the importance of this study is the identification of some of the volatile
bioactive compounds in the leaves of the plant. Further work on the pharmacological activity
of this plant can be considered.

ACKNOWLEDGEMENTS
The authors express appreciation to the Vice Chancellor, Covenant University, the
management team of the Covenant University Centre for Research, Innovation and
Development (CUCRID) for support grant towards this research and to the Chemistry

5
3rd International Conference on Science and Sustainable Development (ICSSD 2019) IOP Publishing
IOP Conf. Series: Journal of Physics: Conf. Series 1299 (2019) 012014 doi:10.1088/1742-6596/1299/1/012014

Department, Ahmadu Bello University for the use of the facilities of their Multi-User
laboratory.

Conflict of Interest: The authors hereby declare that there is no conflict of interest

REFERENCES
[1] Ayepola, O.O and Ishola, R.O. (2009). Evaluation of Antimicrobial Activity of
Anacardiumoccidentale (Linn.)Advances in Medical and Dental Sciences, 3(1): 1-3.
[2] Cragg, G.M. and Newman, D.J. (2005). Plants as a Source of Anti-Cancer Agents.Journal
of Ethnopharmacology, 100, 72-79. http://dx.doi.org/10.1016/j.jep.2005.05.011
[3] Mann, A; Ibrahim, K; Oyewale, A.O; Amupitan, J.O. &Okogun, J.I. (2009).
Antimycobacterial activity of some medicinal plants in Niger State, Nigeria.African
Journal of Infectious Diseases,3(2): 44–48.
[4] Burkill, H.M. (1985).The Useful Plants of West Tropical Africa, vol. 1.Families A – D,
second ed. Royal Botanic Gardens, Kew.
[5] Adebolu, T.T., Adeboye, P.T., and Adegbola, N.B. (2007). Evaluation of a Traditional
Decoction Made from Psidiumguajava and Zingiberofficinale for Anti Bacterial
Activity.Research Journal of Microbiology, 2: 954- 959.
[6] Agbankpe, A. J, Bankoles, H. L, Dougnon, T. J, Yehouenou, B, Hounmanou, Y.M.G,
BabaMoussa, L.S.( 2015). Comparison of Nutritional Values of Vernoniaamygdalina,
Crateva adansonii and Sesamumradiatum: Three Main Vegetables Used in Traditional
Medicine for the Treatment of Bacterial Diarrhoea in Southern Benin (West Africa).
Food and Public Health.5(4): 144-149. DOI:http://dx.doi:10.5923/j.fph.20150504.07
[7] Tsado, A. N.; Bashir, L.; Mohammed, S. S.; Famous, I. O.; Yahaya, A. M.; Shu’aibu, M.;
Caleb, T. (2015). Phytochemical composition and antimalarial activity of methanol leaf
extract of Crateva adansonii in Plasmodium berghei infected mice. British
Biotechnology Journal, 6(4): 165-173.
[8] Adjagba, M., Awede, B., Osseni, R., Hountondji, C., Dougnon, G., Lagnika, L., Darboux,
R., and Laleye, A. (2017). Antihypertensive effect of extracts from Crateva
adansoniiDC.ssp. adansonii in the Wistar rats. International Journal of Biology and
Chemical Sciences, 11(6): 2604-2615.
[9] Ganesan S.; Ponnuchamy M.; Kesavan L.; Selvaraj A. (2009). Floristic composition and
practices on the selected sacred groves of Pallapatty village (Reserved forest), Tamil
Nadu. Indian Journal of Traditional Knowledge, 8(2):154 - 162.

6
3rd International Conference on Science and Sustainable Development (ICSSD 2019) IOP Publishing
IOP Conf. Series: Journal of Physics: Conf. Series 1299 (2019) 012014 doi:10.1088/1742-6596/1299/1/012014

[10] Rathinavelab, T.; Ammashib, S; Muthusamy, G. (2018). Screening of anti-inflammatory

phytocompounds from Crateva adansonii leaf extracts and its validation by in
silicomodeling. Journal of Genetic Engineering and Biotechnology, 16(2); pp 711-719.
[11] Abenga, J.N., Lawal, I.A. (2005). Implicating roles of animal reservoir host in the
resurgence of Gambian trypanosomiasis (Sleeping Sickiness).African Journal of
Biotechnology, 4: 134–137.
[12] Ajanaku, C. O., Echeme, J. O., Mordi, R. C., Owoeye, T. F., Taiwo, O. S., Ejilude, O.,
and Ataboh, J. U. (2018). Phytochemical Screening and Antimicrobial Studies of Stem
and Root Extracts of Crateva adansonii. Annual Research & Review in Biology, 24(5):
1-7.
[13] Kanthal, L. K., Dey, A.,Satyavathi, K. and Bhojaraju, P. (2014). GC-MS analysis of
bio-active compounds in methanolic extract of Lactucaruncinata DC.Pharmacognosy
Research, 6(1): 58-61.
[14] Uduman, M.S.,Rathinam, P.,Karuru, Y.,Obili, G, Chakka, G.,Janakiraman, A.K. (2017).
GC-MS Analysis of Ethyl Acetate Extract of Whole Plant of Rostellulariadiffusa.
Pharmacognosy Journal, 9(1):70-72.
[15] Wagay, N. A. (2017) Investigations On Secondary Metabolites From Crateva
ReligiosaForst.–A Rare Medicinal Plant Of Vidarbha Region (M.S.) India. International
JournalofPharmacetical Biological Sciences, 8(1): 402 – 407
[16] Sacks, F. M and Campos, H. (2006). Polyunsaturated Fatty Acids, Inflammation, and
Cardiovascular Disease: Time to Widen Our View of the Mechanisms, The Journal of
Clinical Endocrinology & Metabolism,91(2):398–400.
[17] Ander, B. P.,Dupasquier, C. M.,Prociuk, M. A. and Pierce, G. N. (2003).
Polyunsaturated fatty acids and their effects on cardiovascular disease.Experimental and
Clinical Cardiology, 8(4): 164-72.
[18] Shahidi, F. andAmbigaipalan, P.(2018). Omega-3 Polyunsaturated Fatty Acids and
Their Health Benefits.Annual Review of Food Science and Technology, 9(1): 345-381
[19] Hu, F.B., Bronner J., Willett W.C., Stampfer M.J., Rexrode K.M., Albert C.M., (2002)
Fish and omega-3 fatty acid intake and risk of coronary heart disease in women. JAMA,
287:1815–1821.
[20] Aparna, V., Dileep, K.V., Mandal, P.K., Karthe, P., Sadasivan, C. andHaridas, M.
(2012). Anti-inflammatory property of n-hexadecanoic acid: structural evidence and
kinetic assessment. Chemical Biology & Drug Design, 80 (3): 434-439.
[21] Sanni, D. M., andOmotoyinbo, O. V. (2016). GC-MS Analysis of Pteleopsissuberosa
Stem Bark Methanol-Chloroform Extract. Journal of Plant Sciences.4(3): 37-40.
[22] Whelan, J. and McEntee, M. F. (2004). Dietary (n-6) PUFA and Intestinal
Tumorigenesis.Journal of Nutrition, 134: 3421S-3426S.

7
3rd International Conference on Science and Sustainable Development (ICSSD 2019) IOP Publishing
IOP Conf. Series: Journal of Physics: Conf. Series 1299 (2019) 012014 doi:10.1088/1742-6596/1299/1/012014

[23] Adeoye-Isijola, M. O., Olajuyigbe O. O., Jonathan S. G., and Coopoosamy R. M.,
(2018). Bioactive Compounds In Ethanol Extract of LentinusSquarrosulus Mont – A
Nigerian Medicinal Macrofungus. African Journal of Traditional Complementary
Alternative Medicine, 15(2): 42-50.