ARID ZONE JOURNAL OF ENGINEERING, TECHNOLOGY & ENVIRONMENT AZOJETE June 2023. Vol. 19(2):355-366 Published by the Faculty of Engineering, University of Maiduguri, Maiduguri, Nigeria. Print ISSN: 1596-2490, Electronic ISSN: 2545-5818 www.azojete.com.ng Corresponding author’s e-mail address: adebayoadewale@ymail.com, adebayow@oauife.edu.ng 341 ORIGINAL RESEARCH ARTICLE CHEMICAL CONSTITUENTS OF SOLVENT EXTRACTED BITTER KOLA NUT OIL AND ITS BENEFCIAL HEALTH PROPERTIES W. A. Adebayo Department of Food Science and Technology, Obafemi Awolowo University, Ile-Ife, Nigeria *Corresponding author’s email address: adebayoadewale@ymail.com, adebayow@oauife.edu.ng 1.0 Introduction Nuts and seed are nutrient dense foods with complex matrices rich in unsaturated fatty acids (60 – 98%), high quality protein, fiber, minerals (iron, potassium, magnesium, calcium, phosphorus and selenium), and other bioactive compounds such as tocopherols, phytosterols, and phenotic compounds (Nzekwe et al., 2015). Prominent nuts and seeds are groundnut, almonds, cashew nuts, cotton seeds, palm kernels, pecans, sesame seeds, soybean seed, sunflower seeds and walnuts. They have been widely utilized in various food formulation and development due to their nutritional and health benefits (Gonzalez-Perez and Arellano, 2009). Vegetable oils extracted from nut and seeds are important ingredients in the food, agro-allied and chemical industries. Prominent vegetable oils are palm oil, soybean oil, canola oil, and sunflower oil which contribute more than 70% of the global vegetable oils consumption (Kojima et al., 2016). The world major exporting countries in vegetable oils are Indonesia, Malaysia, and Argentina while leading importers being India, China and EU (Kojima et al., 2016). Global demand for vegetable oils is greatly affected by the exponential increase in the world’s population, rising awareness of the health benefits of the unsaturated fatty acids present in the vegetable oils and increased industrial usage (Kojima et al., 2016). Recent advances in technology increases global utilization of vegetable oils therefore put pressure on conventional nut and seeds. Therefore, there is dire need to explore non-conventional nuts and seeds. One of the non-convectional nuts and seeds is bitter kola (Garcinia kola). Bitter kola is a species of dicotyledonous flowering plant in the Malphighiales plantae order and Clusiaceae family (Manourova et al., 2019). It is a perennial crop growing in the forest, distributed throughout West and Central Africa where its favorable climatic conditions ARTICLE INFORMATION ABSTRACT This study investigated chemical composition and biological activity of bitter kola oil. Solvent extraction was done using Soxhlet extraction method and the chemical composition of the oil was investigated using standard procedures. The oil yield of bitter kola nut oil was 2.480%. The color was deep brown. The chemical composition of the oil showed 58 different chemical compounds of which 40 are reported to have biological activity. 23.1% of the chemical composition of bitter kola oil obtained was made up of fatty acids. The fatty acid compositions of the oil showed saturated and unsaturated components that ranged from 0.59 – 10.10% and 0.72 – 4.13%, respectively. Some of the saturated components of bitter kola nut oil include Palmitic; Lauric; Myristic; Pentadecanoic and Stearic fatty acids while the unsaturated components were Oleic; Linoleic and Hydnocarpic fatty acids. In conclusion, this study showed that bitter kola oil comprises of many biologically active compounds which could be used could beuse in food formulation, pharmaceutical and chemical industries. © 2023 Faculty of Engineering, University of Maiduguri, Nigeria. All rights reserved. Submitted 12 December, 2022 Revised 27 February, 2023 Accepted 6 April, 2023 Keywords: Chemical constituents solvent extraction bitter kola oil beneficial health http://www.azojete.com.ng/ file:///C:/Users/Engr.%20Samuel/Documents/Engr%20Oyeniyi/azojete/AZOJETE%20ARCHIVE/UPLOAD/VOL%2019%20NO%202/AZOJETE%20VOL%2019%20NO%202/adebayoadewale@ymail.com,%20adebayow@oauife.edu.ng adebayoadewale@ymail.com,%20adebayow@oauife.edu.ng Arid Zone Journal of Engineering, Technology and Environment, June, 2023; Vol. 19(2):355-366. ISSN 1596-2490; e-ISSN 2545-5818; www.azojete.com.ng Corresponding author’s e-mail address: adebayoadewale@ymail.com, adebayow@oauife.edu.ng 356 are met (Iwu, 1993). In Nigeria, bitter kola is primarily cultivated in Southern states such as Ekiti, Osun, Oyo, Ogun, Ondo, Edo and Delta. Bitter kola stem and leaves have been employed over the centuries to treat different ailments but the most valued products of bitter kola are from its nuts, which possess anti-microbial, anti-diabetic and anti-inflammatory properties (Okoye et al., 2014). Eleyinmi et al. (2006) documented that bitter kola nut comprises of crude protein (39.52 g/kg), lipid extract (43.25 g/kg), ash (11.42 g/kg), crude fiber (114.02 g/kg), and a carbon: nitrogen ratio of 57.88. Despite enormous benefits of bitter kola vegetable oil, it is still classified as an underutilized nut. Therefore, there is need to investigate its chemical and biological properties as this will provide scientific data that will promote possible utilization of bitter kola vegetable oil for both domestic and industrial usage. 2. Materials and Methods 2.1 Source of materials Mature bitter kola nut was obtained from Teaching and Research farm, Obafemi Awolowo University, Ile-Ife, Nigeria. All chemicals needed for this work were of analytical (Sigma Chemical (St. Louis, MO, USA and Fisher Scientific, Oakville, ON, Canada) grades. 2.2 Sample preparation About 5 kg of matured bitter kola nut was sorted and washed with potable water to remove foreign particles attached to the nuts. The cleaned bitter kola nut was decorticated, dried in a hot-air oven (AFOS Mini Kiln, Hull, England) and milled with a laboratory milling machine (Retsch GmbH and Co., Haan, Germany). The milled bitter kola nut was then subjected into solvent extraction process using n-hexane (analytical grade) as extraction solvents. Milled bitter kola nut (1.00 kg) was measured and weighed into the extraction thimble of known weight. The loaded thimble was then placed in a Soxhlet apparatus. The extraction solvent (5 L) was measured into the round bottom flask for each extraction process. Then, the temperature of the heating mantle was set to 50 ˚C. As the temperature of the solvent increased, as a result of heat convection, solvent began to vaporize and vapor formed then passed through the thimble and began to condense due to the cooling effect of the water flowing in and out of the Soxhlet extractor setup. This continuous process of heating and cooling was carried out until considerable amount of bitter kola oil was extracted after 8 h. At the end of the experiment, the thimble was removed from the Soxhlet extractor set-up and the rest of the solvent in the apparatus was recovered. Subsequently, a simple distillation process was carried out on the miscella in order to separate bitter kola oil from the solvent (Figure 1). Then, the extracted oil was labeled accordingly. 2.3 Determination of moisture content The moisture content was determined using the oven drying method. 10 g of decorticated bitter kola nut sample were placed inside a moisture can. The sample was then dried in a Uniscope Laboratory Oven set at the temperature of 105 ˚C for 3 h till no change in mass was observed. After drying, the sample was placed inside a dessicator and allowed to cool to room temperature. Then, the sample was weighed and the values were recorded. The moisture content of the bitter kola nuts was then expressed as percentage moisture content (wet basis) using Equation 1: file:///C:/user/Downloads/azojete143/www.azojete.com.ng file:///C:/Users/Engr.%20Samuel/Documents/Engr%20Oyeniyi/azojete/AZOJETE%20ARCHIVE/UPLOAD/VOL%2019%20NO%202/AZOJETE%20VOL%2019%20NO%202/adebayoadewale@ymail.com,%20adebayow@oauife.edu.ng Adebayo: Chemical Constituents of Solvent Extracted Bitter Kola Nut Oil and its Benefcial Health Properties. AZOJETE, 19(2):355-366. ISSN 1596-2490; e-ISSN 2545-5818, www.azojete.com.ng Corresponding author’s e-mail address: adebayoadewale@ymail.com, adebayow@oauife.edu.ng 357 𝑀𝐶(𝑤.𝑏) = 𝑀b − 𝑀c 𝑀b − 𝑀a × 100% (1) Where: M.C (w.b) = moisture content (wet basis); Ma= weight of moisture can; Mb = weight of moisture can plus sample weight before drying and; Mc = weight of moisture can plus sample weight after drying. Bitter kola nuts Solvent Crude extracted bitter kola oil Figure 1: A flow chart showing solvent extraction of bitter kola oil 2.4 Determination of oil yield Solvent extracted bitter kola oil was transferred to a beaker, and then it was heated in a water bath for 2 h, in order to completely evaporate the solvent from it. At the end of the heating process, the volume of the recovered bitter kola oil was measured and expressed as percentage oil yield, using equation 2: 𝑂𝑌% = 𝑊0 𝑊𝑆 × 100 (2) Where: OY = Oil Yield, %; Ws = Weight of milled bitter kola nut used, g W0 = Weight of extracted bitter kola oil, g Sorting Washing Decorticating Drying Milling Soxhlet extractor (n-hexane) http://www.azojete.com.ng/ file:///C:/Users/Engr.%20Samuel/Documents/Engr%20Oyeniyi/azojete/AZOJETE%20ARCHIVE/UPLOAD/VOL%2019%20NO%202/AZOJETE%20VOL%2019%20NO%202/adebayoadewale@ymail.com,%20adebayow@oauife.edu.ng Arid Zone Journal of Engineering, Technology and Environment, June, 2023; Vol. 19(2):355-366. ISSN 1596-2490; e-ISSN 2545-5818; www.azojete.com.ng Corresponding author’s e-mail address: adebayoadewale@ymail.com, adebayow@oauife.edu.ng 358 2.5 Color The color of the extracted bitter kola oil was determined by physical observation. 2.6 Determination of chemical composition of solvent extracted bitter kola oil The chemical components of bitter kola oil such as alkanes, alkenes, alcohols and aromatic hydrocarbons were evaluated using AOAC (2002) methods. First, the oil sample was pre-treated with a process of methylation, following which it was subjected to GC-MS (Gas Chromatography- Mass Spectrometry (Model GL – 15A, Schimadzu Corporation, Kyoto, Japan) analysis. The column temperature of the GC-MS was programmed between 60 ˚C and 300 ˚C. 2.7 Fatty acid profile The fatty acid profile of solvent extracted bitter kola oil was determined by using gas chromatography as a separation method. Fatty acids (FAs) were determined by means of indirect method, which involves their conversion to corresponding methyl esters (fatty acid methyl esters; FAMEs) due to their reaction with trimethylsufonium hydroxide solution. Individual fatty acids were then quantified by means of two distinct methods. Firstly, quantification was based on the received peak area, following which the results were normalized without correction factor. Secondly, quantification was based on the external calibration curve (AOCS, 2009). 2.8 Statistical Analysis Values obtained after all analyses were taken as data, which were then analyzed inferentially with the aid of statistical tools such as mean, analysis of variance (ANOVA), and Tukey test (p < 0.05). 3. Results and Discussion The initial moisture content of the bitter kola nut was 49.67% (w.b) (Table 1). The initial moisture content value obtained is lower than 60.48 and 71.9% reported by Odebunmi et al. (2009) and Onyekwelu et al. (2015), respectively but higher than 15.20% documented by Adeyeye et al. (2017). This wide range of moisture content obtained from various studies could be due to different levels of maturity of bitter kola nut samples, loss of moisture during transportation and storage and different farming practices. The high moisture content value (49.67%, w.b) obtained shows that the nut is susceptible to moisture-dependent deterioration. Also, it shows that bitter kola nut has poor keeping quality unless it is subjected to drying. Table 1: Physico-chemical properties of bitter kola oil Properties HEXSO Initial moisture content Oil yield (%) 49.67% (w.b) 2.480 Physical observation (Colour) Deep brown HEXSO = N-hexane solvent extracted bitter kola oil The percentage oil yield value of bitter kola nut obtained was 2.48% (Table 1). The oil yield value obtained is higher than 2.09% documented by Olatunde et al. (2021) but lower than 8.5% reported by Ordu et al. (2018). These disparities in values of oil yield are attributed to different levels of maturity of bitter kola nut samples and different handling and storage practices. The oil yeild obtained was lower when compared with oil yield of conventional nuts and seeds such as file:///C:/user/Downloads/azojete143/www.azojete.com.ng file:///C:/Users/Engr.%20Samuel/Documents/Engr%20Oyeniyi/azojete/AZOJETE%20ARCHIVE/UPLOAD/VOL%2019%20NO%202/AZOJETE%20VOL%2019%20NO%202/adebayoadewale@ymail.com,%20adebayow@oauife.edu.ng Adebayo: Chemical Constituents of Solvent Extracted Bitter Kola Nut Oil and its Benefcial Health Properties. AZOJETE, 19(2):355-366. ISSN 1596-2490; e-ISSN 2545-5818, www.azojete.com.ng Corresponding author’s e-mail address: adebayoadewale@ymail.com, adebayow@oauife.edu.ng 359 groundnut (40 – 50%), palm kernel (39.53%), sesame (44.80%), rapeseed (40.00%), sunflower (25.50%) and canola (40.00%) (Pahl, 2008; Dim, 2013; Chauchan et al., 2016; Sitepu et al., 2021). Color of bitter kola nut oil by physical observation is deep brown (Table1). Similar observation was documented by Adeyeye et al. (2017). However, Ordu et al. (2018) reported reddish-brown while Olatunde et al. (2021) reported light yellow. These disparities in color of bitter kola oil may be attributed to different levels of maturity of bitter kola nut samples, loss of moisture during transportation and storage, different types of growing soil, different farming practices and different methods of extraction. The color of bitter kola oil is affected by constituents of the nut such as tannins, saponins, terpenoids and flavonoids (Adeyeye et al. 2017). Color is an important parameter used as an indication of adulteration of oils (Lira et al., 2020). The chemical composition of bitter kola nut oil is presented in Table 2. Fifty-eight (58) different chemical compounds are obtained over 68 peaks, using Gas Chromatography- Mass Spectrometry analysis method (GC-MS). Table 2: Chemical composition of bitter kola nut oil Peak No Compound RT %Peak Area 1 .beta.-Myrcene 5.734 0.22 2 Heptane, 3-(chloromethyl)- 5.919 0.08 3 Heptane, 3-(chloromethyl)- 5.970 0.17 4 Benzoic acid 8.123 0.85 5 1-Dodecene 8.242 0.17 6 Dodecane 8.377 0.85 7 Geraniol 8.875 0.12 8 1-Tridecene 10.559 0.62 9 Tetradecane 10.686 2.69 10 Phenol,3,5-bis(1,1-dimethylethyl) 11.689 3.89 11 11-(2-Cyclopenten-1-yl)undecanoic acid, (+)- (Hydnocarpic acid) 11.883 0.72 12 Dodecanoic acid (Lauric acid) 12.289 0.59 13 1-Hexadecanol 12.641 1.08 14 Hexadecane 12.756 3.96 15 2-Bromo-4,6-di-tert-butylphenol 12.807 0.71 16 1-Octadecanesulphonyl chloride 13.554 0.16 17 Tetradecanoic acid (Myristic acid) 14.166 1.24 18 1-Nonadecene 14.507 1.04 19 Heneicosane 14.607 4.01 20 Pentadecanoic acid (Pentadecylic acid) 15.011 0.60 21 7,9-Di-tert-butyl-1-oxaspiro (4,5) deca-6,9- diene-2,8-dione 15.344 0.17 22 1-Octadecanesulphonyl chloride 15.429 0.44 23 Malonic acid, 2-chloropropyl pentadecyl ester 15.461 0.31 24 Hexadecanoic acid, methyl ester 15.527 0.51 25 Dibutyl phthalate 15.667 4.06 http://www.azojete.com.ng/ file:///C:/Users/Engr.%20Samuel/Documents/Engr%20Oyeniyi/azojete/AZOJETE%20ARCHIVE/UPLOAD/VOL%2019%20NO%202/AZOJETE%20VOL%2019%20NO%202/adebayoadewale@ymail.com,%20adebayow@oauife.edu.ng Arid Zone Journal of Engineering, Technology and Environment, June, 2023; Vol. 19(2):355-366. ISSN 1596-2490; e-ISSN 2545-5818; www.azojete.com.ng Corresponding author’s e-mail address: adebayoadewale@ymail.com, adebayow@oauife.edu.ng 360 26 n-Hexadecanoic acid (Palmitic acid) 15.955 10.10 27 m-Cymene,5-tert-butyl 16.033 1.63 28 n-Nonadecanol-1 16.201 1.94 29 Eicosane 16.290 3.45 30 1,6,10,14-Hexadecatetraen-3-ol,3,7,11,15- tetramethyl-(E,E) 16.417 0.72 31 Kaur-16-ene 16.781 0.22 32 n-Heptadecanol-1 16.875 0.50 33 9-Octadecenoic acid, methyl ester, (E)- 16.999 0.23 34 Octadecane,1-chloro- 17.233 0.64 35 9,12-Octadecadienoic acid (Z,Z) (Linoleic acid) 17.408 3.92 36 9-Octadecenoic acid, (E)- (Oleic acid) 17.490 4.13 37 Octadecanoic acid (Stearic acid) 17.698 1.80 38 trans-Geranylgeraniol 17.875 0.87 39 1-Hydroxy-1,7-dimethyl-4-isopropyl-2,7- cyclodecadiene 17.951 0.38 40 9-Hexadecenoic acid, octadecyl ester, (Z)- 18.068 1.10 41 Heneicosane 18.167 1.84 42 1,1,6-trimethyl-3-methylene-2-(3,6,9,13- tetramethyl-6-thenye-10,14-dimethylene- pentadec-4-enyl) cyclohexane 18.490 0.75 43 17-Pentatriacontene 19.376 0.63 44 9-Octadecenamide, (Z) 19.455 1.44 45 [1,1’-Biphenyl]-2,3’-diol,3,4’,5,6’-tetrakis (1,1- dimethylethyl 19.540 0.81 46 Behenic alcohol 20.029 0.27 47 Eicosane 20.108 1.01 48 Phosphoric acid, tris (2-ethylhexyl) ester 20.587 1.33 49 Trichloroacetic acid, undecyl ester 20.791 1.09 50 Eicosane 20.994 0.77 51 Bis (2-ethylhexyl) phthalate 21.121 5.09 52 1H-Pyrazole-5-carboxamide,3-cyclopropyl-N- (3,5-dimethyltricyclo [3.3.1.1(3,7)] dec-1-yl)- 1-methyl- 21.542 0.22 53 2-methylhexacosane 21.807 0.68 54 Oleoyl chloride 21.252 1.48 55 Behenic alcohol 21.433 1.04 56 Eicosane 22.575 0.85 57 Methoxy-THC 22.620 0.42 58 9-Octadecenamide, (Z)- 22.906 0.37 59 Decanedioic acid, bis (2-ethylhexyl) ester 23.177 0.75 60 Eicosane 23.313 0.29 61 Squalene 23.433 7.11 62 1-Heptacosanol 23.955 4.10 63 Octadecane,3-ethyl-5-(2-ethylbutyl)- 24.056 1.78 64 trans-Geranylgeraniol 24.165 0.51 file:///C:/user/Downloads/azojete143/www.azojete.com.ng file:///C:/Users/Engr.%20Samuel/Documents/Engr%20Oyeniyi/azojete/AZOJETE%20ARCHIVE/UPLOAD/VOL%2019%20NO%202/AZOJETE%20VOL%2019%20NO%202/adebayoadewale@ymail.com,%20adebayow@oauife.edu.ng Adebayo: Chemical Constituents of Solvent Extracted Bitter Kola Nut Oil and its Benefcial Health Properties. AZOJETE, 19(2):355-366. ISSN 1596-2490; e-ISSN 2545-5818, www.azojete.com.ng Corresponding author’s e-mail address: adebayoadewale@ymail.com, adebayow@oauife.edu.ng 361 65 1-(2,5-Bis-methosymethoxy-phenyl)-3,7,11- trimethyl-dodeca-2,6,10-triene 24.240 0.66 66 Oct-5-en-2-ol, 8-(1,4,4a,5,6,7,8,8a-octahydro- 2,5,5,8a-tetramethylnaphth-1-yl)-6-methyl- 24.587 1.71 67 Purine-2,6-dione,1,3-dimethyl-7-(2-oxo-2- phenylethyl)-8-(piperidin-1-yl)-3,7-dihydro 24.697 1.35 68 Ethyl trans-4a, cis-4b,trans-8a,cis-10a- perhydro-trans-2,4a,8a-trimethyl-8- oxophenanthrene-2-carbothiolate 24.864 0.78 RT: Retention time The compound with the highest concentration is n-Hexadecanoic acid (Palmitic acid) (10.10%), a saturated fatty acid with reported anti-inflammatory, anti-oxidant and anti-microbial biological activities (Abubakar and Majinda, 2016). Other identified compounds with relatively high concentrations were: Squalene (7.11%), Eicosane (6.37%), Heneicosane (5.85%), Bis(2- ethylhexyl)phthalate (5.09%), 9-Octadecenoic acid (4.13%), 1-Heptacosanol (4.10%), Dibutyl phthalate (4.06%), Hexadecane (3.96%), 9-12-Octadecadienoic acid(Z,Z) (Linoleic acid) (3.92%), Phenol, 3,5-bis(1,1-dimethylethyl) (3.89%), and Tetradecane (2.69%); while the compound with the least concentration is Geraniol (0.12%), a fatty alcohol with reported anti-microbial, anti- diabetic, anti-inflammatory, anti-oxidant, cardio-protective, anti-depressant and insecticidal biological activities (Lira et al., 2020). Total percentage of fatty acids obtained is 23.10%. This includes saturated fatty acids (14.33%): Palmitic acid, C16:0 (10.10%); Lauric acid, C12:0 (0.59%); Myristic acid, C14:0 (1.24%); Pentadecanoic acid, C15:0 (0.60%) and Stearic acid, C18:0 (1.80%). Palmitic acid (10.10%) is the prominent saturated fatty acid, a fatty acid with anti-microbial, anti- inflammatory and anti-oxidant properties used in food production, soap production, skincare, cosmetics and other industries (Abubakar and Majinda, 2016) while Lauric acid (0.59%), a fatty acid with anti-microbial and anti-viral activities (Lira et al., 2020), has the least concentration among the saturated fatty acids although Adeyeye et al. (2017) documented higher concentration of total saturated fatty acids (25.74%), with lower percentage of Palmitic acid (5.50%) and higher percentage of Lauric acid (2.57%). Total percentage of unsaturated fatty acids is 8.77% which includes: Hydnocarpic acid, C16:1 (0.72%), Linoleic acid, C18:2 (3.92%) and Oleic acid, C18:1. The unsaturated fatty acid with the highest concentration is Oleic acid (4.13%), a monounsaturated fatty acid with antimicrobial activity, cholesterol lowering capacity and reported health benefits (Lira et al., 2020), while the least concentration of unsaturated fatty acids is Hydnocarpic acid (0.72%); another monounsaturated fatty acid with anti-oxidant properties which is traditionally used to treat leprosy (Lira et al., 2020). Although, Adeyeye et al. (2017) documented a higher concentration of total unsaturated fatty acids (74.26%), with higher percentage of Oleic acid (24.23%). Also, total percentages of mono-unsaturated and poly- unsaturated fatty acids were 4.85 and 3.92%, respectively. The ratios of poly-unsaturated fatty acid to total saturated fatty acid (P/S) and oleic to linoleic are 0.27 and 1.05, respectively. Although, Adeyeye et al. (2017) reported higher P/S ratio (1.91) and lower oleic/linoleic rato of 0.57; which indicates that bitter kola oil they obtained was not as stable and suitable for frying compared to the oil obtained in this study. These differences in fatty acid profile could be as a http://www.azojete.com.ng/ file:///C:/Users/Engr.%20Samuel/Documents/Engr%20Oyeniyi/azojete/AZOJETE%20ARCHIVE/UPLOAD/VOL%2019%20NO%202/AZOJETE%20VOL%2019%20NO%202/adebayoadewale@ymail.com,%20adebayow@oauife.edu.ng Arid Zone Journal of Engineering, Technology and Environment, June, 2023; Vol. 19(2):355-366. ISSN 1596-2490; e-ISSN 2545-5818; www.azojete.com.ng Corresponding author’s e-mail address: adebayoadewale@ymail.com, adebayow@oauife.edu.ng 362 result of different levels of maturity of bitter kola nut samples, loss of moisture during transportation and storage, different types of growing soil and different farming practices. The biological activities of chemical compound of bitter kola oil are presented in Table 3. Forty (40) chemical compounds found in bitter kola oil are reported to have biological activities such as anti-oxidant, anti-inflammatory, anti-microbial, anti-fungal, anti-diabetic, anti-cancer, anti- tuberculosis, anti-depressant, cardio-protective, anti-septic, anti-viral, anti-aging, anti- tuberculosis, anti-epileptic, anti-cytotoxic, psychoactive and insecticidal activity. The anti- microbial effect of bitter kola oil may be attributed to the long aliphatic chains and fatty acid ester in it, which have been reported to build up in the cell membrane and mitochondria of a cell, thus resulting in disruption of the cell integrity and the death of the cell (Belakhdar et al., 2015). Table 3: Biological activities of compounds in bitter kola nut oil Peak No Compound %Peak area Class of compound Biological activity 1 .beta.-Myrcene 0.22 Acyclic alkene Anti-oxidant, anti-aging, anti-inflammatory 4 Benzoic acid 0.85 Aromatic hydrocarbon Anti-microbial 5 1-Dodecene 0.17 Acyclic alkene Anti-bacterial 6 Dodecane 0.85 Acyclic alkane Anti-oxidant 7 Geraniol 0.12 Terpene alcohol Anti-microbial, anti- diabetic, anti- inflammatory, anti- oxidant, cardio- protective, anti- depressant, insecticidal 8 1-Tridecene 0.62 Acyclic alkene Pheromone and allomone properties 9 Tetradecane 2.69 Acyclic alkane Anti-microbial, anti- tuberculosis 10 Phenol,3,5-bis(1,1- dimethylethyl) 3.89 Aromatic hydrocarbon Anti-oxidant 11 11-(2-Cyclopenten-1- yl)undecanoic acid, (+)- (Hydnocarpic acid) 0.72 Fatty acid Anti-oxidant 12 Dodecanoic acid (Lauric acid) 0.59 Fatty acid Anti-microbial 13 1-Hexadecanol 1.08 Fatty alcohol Anti-microbial and anti- oxidant 14 Hexadecane 3.96 Acyclic alkane Anti-microbial and anti- oxidant 17 Tetradecanoic acid (Myristic acid) 1.24 Fatty acid Larvicidal and repellent activity 18 1-Nonadecene 1.04 Acyclic hydrocarbon Anti-fungal 19 and 41 Heneicosane 5.85 Alkane Anti-microbial file:///C:/user/Downloads/azojete143/www.azojete.com.ng file:///C:/Users/Engr.%20Samuel/Documents/Engr%20Oyeniyi/azojete/AZOJETE%20ARCHIVE/UPLOAD/VOL%2019%20NO%202/AZOJETE%20VOL%2019%20NO%202/adebayoadewale@ymail.com,%20adebayow@oauife.edu.ng Adebayo: Chemical Constituents of Solvent Extracted Bitter Kola Nut Oil and its Benefcial Health Properties. AZOJETE, 19(2):355-366. ISSN 1596-2490; e-ISSN 2545-5818, www.azojete.com.ng Corresponding author’s e-mail address: adebayoadewale@ymail.com, adebayow@oauife.edu.ng 363 20 Pentadecanoic acid (Pentadecylic acid) 0.60 Fatty acid Anti-microbial 24 Hexadecanoic acid, methyl ester 0.51 Fatty acid ester Anti-microbial 25 Dibutyl phthalate 4.06 Phthalate ester Anti-microbial 26 n-Hexadecanoic acid (palmitic acid) 10.10 Fatty acid Anti-inflammatory, anti- oxidant, anti-microbial 28 n-Nonadecanol-1 1.94 Fatty alcohol Anti-bacterial 29, 47, 50,56 and 60 Eicosane 6.37 Acyclic alkane Anti-microbial 32 n-Heptadecanol-1 0.50 Fatty alcohol Anti-bacterial 33 9-Octadecenoic acid, methyl ester, (E)- 0.23 Fatty acid methyl ester Anti-microbial 35 9,12-Octadecadienoic acid (Z,Z) (Linoleic acid) 3.92 Fatty acid Anti-oxidant activity 36 9-Octadecenoic acid, (E)- (Oleic acid) 4.13 Fatty acid Anti-microbial 37 Octadecanoic acid (stearic acid) 1.80 Fatty acid Anti-oxidant activity 59 Decanedioic acid, bis (2- ethylhexyl)ester 0.75 Fatty acid ester Anti-microbial 61 Squalene 7.11 Aromatic hydrocarbon Anti-oxidant and anti- cardiovascular disease 62 1-Heptacosanol 4.10 Fatty alcohol Anti-microbial, anti- oxidant 63 Octadecane,3-ethyl-5-(2- ethylbutyl)- 1.78 Acyclic alkane Anti-microbial The biologically active compounds with relatively high concentration are n-Hexadecanoic acid (Palmitic acid) (10.10%), which has reported anti-inflammatory, anti- oxidant and anti-microbial properties (Abubakar and Majinda, 2016); Eicosane (6.37), Heneicosane (5.85%), Dibutyl phthalate (4.06%) and Hexadecane (3.96%) which have reported anti-microbial activity (Usha et al., 2015; Ahsan et al., 2017; Kim, 2020). Although, there is dearth of information on the biological activity of Kaur-16-ene, its derivatives have been reported to have anti-inflammatory activity (Chavan et al., 2010). Hexadenoic acid methyl ester, 9-Octadecenoic acid methyl ester and 9,12- Octadecadienoic acid were also documented by Fapohunda et al. (2017) although, they occurred in higher concentrations of 4.08, 12.42 and 5.04% respectively. These differences in chemical composition of bitter kola nut oil could be as a result of different levels of maturity of bitter kola nut samples, loss of moisture during transportation and storage, different types of growing soil and different farming practices. http://www.azojete.com.ng/ file:///C:/Users/Engr.%20Samuel/Documents/Engr%20Oyeniyi/azojete/AZOJETE%20ARCHIVE/UPLOAD/VOL%2019%20NO%202/AZOJETE%20VOL%2019%20NO%202/adebayoadewale@ymail.com,%20adebayow@oauife.edu.ng Arid Zone Journal of Engineering, Technology and Environment, June, 2023; Vol. 19(2):355-366. ISSN 1596-2490; e-ISSN 2545-5818; www.azojete.com.ng Corresponding author’s e-mail address: adebayoadewale@ymail.com, adebayow@oauife.edu.ng 364 4. Conclusion This study shows the chemical composition and biological activity of bitter kola nut oil. Chemical compounds such as n-Hexadecanoic acid (Palmitic acid), Eicosane Heneicosane, Dibutyl phthalate and Hexadecane which occur in relatively high concentration have notable anti-inflammatory, anti-oxidant and anti-microbial properties. Also, other compounds such as 1-Dodecene, n- Nonadecanol and n-Heptadecanol-1 have anti-bacterial activity. Behenic alcohol has anti-viral activity, Squalene has anti-cancer activity and Geraniol, which has the lowest concentration, has anti-depressant, cardio-protective and anti-diabetic activity. 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