ACTA BOT. CROAT. 78 (1), 2019 17 Acta Bot. Croat. 78 (1), 17–24, 2019 CODEN: ABCRA 25 DOI: 10.2478/botcro-2019-0002 ISSN 0365-0588 eISSN 1847-8476 Morpho-chemical divergence and fatty acid profile of shea tree seeds (Vitellaria paradoxa) collected from different locations in Kwara State, Nigeria David Adedayo Animasaun1*, Stephen Oyedeji1, Kehinde Stephen Olorunmaiye1, Musibau A. Azeez2*, Idowu Abdulfatah Tijani3, Joseph Akintade Morakinyo1 1 Department of Plant Biology, Faculty of Life Sciences, University of Ilorin, Ilorin, Kwara State, Nigeria 2 Department of Pure and Applied Biology, Ladoke Akintola University of Technology, Ogbomoso, Oyo State, Nigeria 3 Department of Chemical Engineering, University of Ilorin, Ilorin, Kwara State, Nigeria Abstract – The present study characterizes seed-related traits, phytochemical, physiochemical parameters and fatty acid profile of shea (Vitellaria paradoxa) seeds collected from the Kosubosu, Fufu and Sare areas of Kwara State, Nigeria to determine the effects of microclimate on seed morphology, biochemical and oil constituents. Seed morphological data were analyzed for variability. Seed oil was extracted for phytochemical constituents, physicochemical properties, and fatty acid profiling by gas chromatography equipped with mass spectrometry (GC/MS). Results showed intra and inter-locational variations in seed characters. Most fruits had 1–2 seeds. Seeds were predominantly brown and very few were dark brown. Phytochemicals and physicochemical parameters of the seed oil varied with place of collection. Alka- loid, saponin, tannin and phytate contents ranged between 0.79–0.84, 1.20–1.26, 1.48–1.56 and 0.15–0.18 mg g–1 respec- tively. The density of the oil was less than that of water, acid value ranged from 10.58–13.56 mg KOH g–1 and iodine values were between 36.63 to 40.32 g I2 (100 g)–1. Saponification values lie between 160.39 and 184.14 mg KOH g–1; and free fatty acid was within 5.32–6.81 %. Peroxide, ɑ-tocopherol, total phenol and oxalate values as well as viscosity of the oil also varied; however, refractive index was similar. Ethyl oleate and octadecanoic acids were present and most abun- dance in all the locations, while glycidol stearate was only found in Fufu samples with three other fatty acids. Five fatty acids were present in Kosubosu, while Sare had only two. The results obtained in the present study indicate that shea oil could be used for medicinal, nutritional and industrial purposes. Since seed characters, phytochemical, physicochemi- cal and fatty acid compositions varied with the microclimate, environmental and micro-ecological conditions should be considered when collecting seeds for oil utilization. Keywords: fatty acids, phytochemical analysis, physicochemical parameters, shea butter, Vitellaria paradoxa * Corresponding authors: animasaun.ad@unilorin.edu.ng, musibau_azeez@yahoo.com Introduction Vitellaria paradoxa Gaertn. F., commonly known as the shea tree is a tree of the Sapotaceae family indigenous to Sub-Sahara Africa where it grows in the wild and has huge economic and ecological potentials (Ademola et al. 2012). The tree occurs in West African and is popular among the local dwellers for its numerous social and economic utiliza- tions (Kobomoje et al. 2013). The shea tree grows wild across a wide belt of savanna including West African countries of Senegal, Mali, Cote d’ Ivoire, Burkina Faso, Togo, Ghana, Benin, Nigeria, Niger and Cameroon. The tree is also found further to the east, in Uganda, Sudan and Ethiopia (Maranz et al. 2004, Goreja 2004, Masters et al. 2010). Among these countries, Ghana and Burkina Faso are the main Shea net ex- porters (Walter et al. 2003). In Nigeria, the shea tree is known as “Emi” among the Yoruba people of the western part. The shea grows up to 9–12 m in height with profuse branch- es. Commercial fruiting quantity usually commences at ap- proximately 20 years and the tree can continuously produce shea fruit for many more years (Alander 2004). Late matu- rity of the tree has been the bane of its commercial plantation and shea butter related industries depend on nuts collected from scattered, naturally growing shea trees. In Nigeria, the ANIMASAUN D. A., OYEDEJI S., OLORUNMAIYE K. S., AZEEZ M. A., TIJANI I. A., MORAKINYO J. A. 18 ACTA BOT. CROAT. 78 (1), 2019 shea is found in the Guinea savanna with high tree concen- trations in Niger, Kwara, Kebbi and Kaduna states (Warra 2011). Shea trees blossom between February to March, and the fruit matures and falls in May-June. The fruit is ellipitic, yellow green or light green, about 5–8 cm in circumference and similar to the fig (Chalfin 2004). The fruit has a butter- like, mucous pericarp covering an oval brown or light brown seed surrounded by a fragile shining shell with a large hilum on a broad base (Olaniyan et al. 2007). Usually, a shea fruit contains one seed but occasionally has two to three oil-rich nuts (Alander 2004). The shea tree has the potential to improve nutrition, boost healthcare, reduce rural poverty and support sustain- able land care (Moore, 2008). The fruit pulpy pericarp can be eaten while the nut is discarded or crushed for shea but- ter extraction. The fat obtained from the shea nut known as shea butter (Ori: Yoruba; Nigeria), which contains 40–60% oil, is the most valued product from the shea tree (Warra and Komo, 2014). Shea butter is used for local domestic pur- poses such as cooking, lightning, soap making, skin mois- turizer and cosmetics as well as traditional medicine among the local population where the tree grows in abundance in Nigeria. Also, it is used to treat wounds, suppress inflam- mation, relieve rheumatic and joint pains and other health conditions (Kraft and Lynde 2005, Soro et al. 2011, Warra and Komo 2014). The potential of shea oil to protect the skin from damaging ultra violet rays was demonstrated by Velasco et al. (2008). On a global scale, the importance of the shea tree is related to the usefulness of its seed fat in food and cosmetic industries. Decolourised shea butter could be used as a cheaper alternative to cocoa butter due to their similar physical and chemical properties (Lipp et al. 2001, Alander 2004). The high levels of unsaponifiable matter in shea nut oil compared to other vegetable fats could be exploited for the development of more shea butter products (Rogers and Lenick 2009). Shea oil finds applications in traditional and social rituals such as marriages, funerals, coronations and rainmaking (Ferris et al. 2004, Gwali et al. 2011, Hall et al. 1996, Moore 2008). Also, high grade charcoal and furniture are made from the wood while the latex yields a consider- able amount of glue (Lovett and Haq 2000). Consequently, the shea tree could generate significant incomes for the ru- ral households. However, despite the enormous economic and healthcare potentials, shea butter and its products are underutilised in Nigeria and only little information is available on the seed variability. In particular, not much work has been done on locational variation of seed characters in relation to the nut phytochemicals and fatty acid characterization of seeds from different locations. The present work was carried out to char- acterize seed related traits, quantify phytochemical constitu- ents of the nut and profile the fatty acids of the shea seed col- lected from different locations in Kwara State, Nigeria. This study hopes to identify existing variations in the shea seeds from different locations using seed and oil characteristics to throw more light on how microclimate affects seed morphol- ogy and chemical constituents in shea tree. Materials and methods Fruit and seed collection Vitellaria paradoxa fruits were collected from three lo- cations in Kwara State, Nigeria, because of the abundance of shea trees, the folkloric processing and utilization of shea but- ter for socio-economic purposes. Kwara State is located in the Southern Guinea savanna belt of Nigeria, which is dominated by grasses interspersed mainly with tree species such as Dani- ellia oliveri, Vitellaria paradoxa, Prosopis africana and Parkia biglobosa. The state is within the coordinates 8°30'0.00"N and 4°32'32"E and at an elevation of 303 m above sea level (NGIA, 2016). The names and coordinates of the three locations from where shea fruits and seeds are collected are shown in Table 1. The soil was mainly sandy, lateritic or ferralitic, mean annual rainfall is 640–1350 mm. Usually, the rainy season lasts from May to October and dry season from November to April. Rel- ative humidity is 75–80% in the wet season and about 65% during the dry season (Ajadi et al. 2011). The average month- ly temperature is 31.4–33.5 °C, but fluctuates between 33 and 34 °C from November to January and 34–35 °C (February to April) (Ilorin Atlas, 1982; NIMET, 2016). Tab. 1. Names, local division, coordinates and elevations of the locations from where shea fruits and seeds were collected for the study in Kwara State of Nigeria. Location name Local government area Coordinates Altitude (m) Kosubosu Baruten 9°35'N, 3°15'E 273-364 Fufu Ilorin West 8°30'N, 4°32'E 300-310 Sare Ifelodun 9°40'N, 3°22'E 370-418 Fruit and seed morphological studies Ten mature fruit bearing trees were selected at each lo- cation (Kosubosu, Fufu and Sare) from May to July, 2016. Each location has five sites from which two fruiting trees were selected. Thirty ripe fruits were randomly plucked from each tree for characterization. Epicarp thickness and seed di- ameter were determined using an electronic vernier calliper (ATD-8656). Number of seed per fruit, seed dimension and seed coat colour were also recorded; the mean values of ten readings were used for the seed characters. Shea nuts oil extraction The pulpy pericarp of the fruits were removed, the seeds cleaned and sun dried for two weeks. The dried seeds were deshelled, the kernels were ground and kept in airtight con- tainers prior to oil extraction. Oil extraction was carried out in Soxhlet apparatus using n-hexane as solvent. 50 g of the ground material was transferred to a 30 mm × 200 mm cel- lulose thimble placed in the extraction chamber of a 250 mL Soxhlet apparatus fitted with a condenser on a 500-mL dis- tillation flask containing 250 mL of n-hexane solvent. Shea nut oil was extracted under reflux with n-hexane following VARIATION IN SHEA SEEDS FROM DIFFERENT LOCATIONS ACTA BOT. CROAT. 78 (1), 2019 19 the procedure described by Ali et al. (2015). After extraction, hexane was removed by using a heated rotary evaporator (Stuart, England), under vacuum conditions to recover the oil. Extractions of oil from seeds of different locations were performed in triplicate, and the mean values were reported. Oil yield was expressed as percentage weight of oil obtained relative to the weight of shea butter used for extraction ac- cording to Warra et al. (2011). Phytochemical and physicochemical properties determination Methanolic extracts of the shea nut were screened for presence or otherwise of bioactive compounds using stan- dard procedures (Sofowora 1993, Kokate 1999, Kumaran and Karunakaran 2006). The amount of tannin was estimated using the method described by Hagerman et al. (2000), sa- ponin, and alkaloid present in the shea nut extract were de- termined according to procedure of Obadoni and Ochuko (2001). Total phenol, oxalate and phytate contents in the extract were quantified according to the methods of Yadav et al. (2011) and Falana et al. (2016), while the amount of α-tocopherol was determined by the standard procedure of he Association of Official Analytical Chemists (2010). Iodine values (IV) of the shea oil were estimated using the method of Gafar et al. (2012). The value was calculated from fatty acid methyl ester compositions of oil. Saponifica- tion value was calculated from fatty acid methyl ester com- positions of oil using the equation of Kalayasiri et al. (1996). Determination of the acid value, which is the amount of car- boxylic acid groups in the oil, was carried out based on a titration method using the AOAC (2010) procedure. Vis- cosity of the shea butter oil was recorded using an Ostwald viscometer (DV-III ultra, UK). The free fatty acid of the oil was estimated by placing 0.2 g of oil into 250 cm3 Erlenmeyer flask, 100 cm3 of ethanol was added and followed by 2 cm3 of phenolphthalein indicator. The mixture was titrated against 0.1 M NaOH till endpoint (slight pink colour that persists for 30seconds), the free fatty acid was calculated according to Chopra and Kanwar (1991). To determine the peroxide value of the oil, 2.0 g of oil was added to 22 cm3 of a solution containing 12 cm3 chloroform and 10 cm3 acetic acid. 0.5 cm3 saturated potassium iodide was added and allowed to stay with intermittent agitation for 1 minute before 30 cm3 distilled water was added. The mixture was titrated against 0.1 M of Na2S2O3 in the presence of starch indicator until the blue colour had just disappeared. The peroxide value deter- mination was carried out in accordance with Neilson (2003). Refractive index of the oil with reference to air was mea- sured with the Abbe Refractometer (Mettler-Toledo, USA). The specific gravity of the oil relative to water was measured by hydrometer. Fatty acid profiling Fatty acid composition of the oil was determined at the Chemical Engineering Laboratory, University of Ilorin, Ni- geria by a gas chromatograph equipped with a mass spec- trometer (GC/MS; Agilent Technologies; Model: 7890A) with HP column, 30 cm long × 0.32 mm ID × 0.25 μm thick- ness film (SGE, Australia). The GC is fitted with Hewlett Packard (USA) flame ionization detector (FID) and 99.9% helium was used as carrier gas. The oil triacylglycerides (TG) was converted to fatty acid methyl ester (FAME) to decrease the boiling point. 0.2 mL of biodiesel was added to a 1 mL mixture consisting of hexane and 2-propanol (4/5 volume ratio). Before charging the oil, the samples were purified by filtration using 0.2 μm polytetrafluoroethylene syringe micro filter, then 1 μL of the sample was injected into the GC/MS using a 5 μL micro syringe (SGE, Australia). The gas chro- matography condition was continuous flow mode. The in- let temperature was 250 °C while the oven temperature was 250 °C. The injection volume was 1 µL and the split ratio 1:50. The procedure was run for 20 minutes with an average veloc- ity of 37.789 cm s–1 at 17.04 psi pressure. On the other hand, the mass spectrometry was run under transfer line tempera- ture of 240 °C, source temperature at 220 °C. The solvent de- lay was for 2 minutes while the scan mass range was 45–500. Data analysis was conducted with GC/MSD ChemStation In- tegrator software. Results The fruits and seeds of the shea tree collected from three selected local government areas of Kwara State, Nigeria var- ied in number of seed/fruit, pericarp thickness, seed coat co- lour and seed dimension as well as phytochemical and fatty acid constituents. Among the 300 shea fruits (30/tree) col- lected from Kosubosu, Baruten Local Government Area, 265 (88.33%) had 1 seed/fruit, 34 were 2-seeded and only one fruit had 3 seeds (Tab. 2). The average ratios of seeds to fruit were 26.5, 3.4 and 0.1 for 1-seed, 2-seed and 3-seed fruits respectively. In addition, 164 of the seeds had brown seed coats, 112 were light brown while 24 were dark brown. The fruits collected from Fufu (Ilorin West Local Govt Area) are predominantly one seeded. 254 (84.66%) of the fruits were 1-seed, 42 had 2 seeds per fruit and 4 fruits were found with 3 seeds each (Tab. 2). The seed colour distribution showed 129 light brown seeds, 121 brown and 50 dark brown seed coats. Similarly, 1-seed fruits occurred most among the fruits col- lected from Sare (Ifelodun Local Govt Area). 252 out of the 300 sampled fruits had 1 seed, 2 seeds per fruit were found in 44 and 3 seeds occurred in 5 fruits. Brown seeds were the most frequent (160) followed by light brown (119) and the few (21) dark brown seeds were from Sare samples. The mean epicarp thickness ranged from 2.77–3.48 mm in fruits collected from Kosubosu (Fig. 1a). Mean seed diameter and circumference ranged between 2.41 to 3.95 cm and 6.44 to 7.46 cm, respectively. Similarly, seed lengths among the Ko- subosu seed population were alike. Considering fruit and seed characters, Fufu fruits had mean pericarp thickness between 2.88–3.36 mm (Fig. 1b). The seed diameter and circumference were smaller than those of Kosubosu and ranged between 2.41 and 87 cm and 5.66 and 7.10 cm respectively. The greatest seed length was 4.96 cm while the least was 3.88 cm. Seed diam- ANIMASAUN D. A., OYEDEJI S., OLORUNMAIYE K. S., AZEEZ M. A., TIJANI I. A., MORAKINYO J. A. 20 ACTA BOT. CROAT. 78 (1), 2019 nol and alkaloids were found in nuts from Kosubosu which invariably had the least amount of ɑ-tocopherol. The or- der of the amount of ɑ-tocopherol in the samples was Sare>Fufu>Kosubosu while saponin content was highest (1.26 mg g–1) in samples from Sare, followed by Kosubo- su (1.21 mg g–1) and the least (1.20 mg g–1) in Fufu. In con- trast, Fufu samples produced the highest amount of oxalate (0.82 mg g–1) as against 0.80 mg g–1 from Kosubosu and 0.78 mg g–1 obtained from Sare samples. eter and circumference were similar for the fruits collected from 10 different trees in Sare, but pericarp thickness and seed length varied (Fig. 1c). The fruits and seeds are smaller than those of Kosubosu and Fufu areas. The average pericarp thickness, seed diameter and length were between 2.61 and 3.37 mm, 2.47 and 2.66 cm, 5.02 and 5.82 cm respectively, however, seed circumference varied from 7.02–7.70 cm. The phytochemical compositions of the shea nut are presented in Table 3. Highest phytate, tannin, total phe- Fig. 1. Morphological seed related characters of shea tree seeds collected from different locations in Kwara State, Nigeria: (a) Kosubosu, Baruten, (b) Fufu, Ilorin West, (c) Sare, Ifelodun. Tab. 2. Morphological characteristics of shea tree seeds collected from selected locations in three local government areas of Kwara State, Nigeria. TN – number of tree, NFS – number of seeds collected, DB- dark brown, LB- light brown, B – brown. TN NFS Kosubosu Fufu Sare No of seed/fruit Seed colour No of seed/fruit Seed colour No of seed/fruit Seed colour 1 2 3 DB LB B 1 2 3 DB LB B 1 2 3 DB LB B 1 30 29 1 0 13 17 22 7 1 6 13 11 13 16 2 1 13 16 2 30 24 6 0 7 11 12 26 3 1 0 22 8 29 1 0 3 10 17 3 30 30 0 0 3 3 24 28 2 0 0 14 16 28 2 0 3 7 20 4 30 30 0 0 4 9 17 30 0 0 25 4 1 29 0 1 4 9 17 5 30 30 0 0 0 15 15 23 7 0 3 10 17 25 5 0 0 14 16 6 30 14 15 1 10 5 15 26 4 0 1 4 25 28 2 0 3 8 19 7 30 29 1 0 0 21 9 28 2 0 0 19 11 21 8 1 1 21 8 8 30 28 2 0 0 19 11 24 5 1 4 18 8 26 3 1 2 18 10 9 30 22 8 0 0 3 27 28 2 0 7 14 9 23 7 0 0 8 22 10 30 29 1 0 0 13 17 19 10 1 4 11 15 30 0 0 4 11 15 Ʃ 300 265 34 1 24 112 164 254 42 4 50 129 121 252 44 5 21 119 160 mean 30 26.5 3.4 0.1 2.4 11.2 16.4 25.4 4.2 0.4 5.0 12.9 12.1 25.2 4.4 0.5 2.1 11.9 16.0 VARIATION IN SHEA SEEDS FROM DIFFERENT LOCATIONS ACTA BOT. CROAT. 78 (1), 2019 21 Percentage oil yield of the shea nut was highest (53.06%) for Kosubosu, followed by Sare (49.26%) and the least in Fufu (47.31%) (Tab. 4). The specific gravity and refractive index of the oil from the different locations were similar, but the least acid value, iodine value and free fatty acid were recorded for Kosubosu samples. Saponification value was higher (184.14 mg KOH g–1) in the Sare sample than in the Kosubosu (182.15 mg KOH g–1) but a much lower value was obtained for Fufu (160.39 mg KOH g–1) which invari- ably had the highest viscosity. Furthermore, the order of peroxide values in the shea oil was Kosubosu>Sare>Fufu (Tab. 4). Table 5 shows the retention time, types and percentage composition of the fatty acids present in shea nut oils. Five fatty acids were found in Kosubosu samples, four in Fufu and only two in Sare. 2-hydroxyl-1(hydroxymethyl) ethyl ester with percentage compositions of 38.89 and 47.43% was the most abundant fatty acid in Kosubosu and Fufu samples re- spectively; it was however not found in Sare samples. Ethyl oleate constituted 59.96% of the total fatty acids in the Sare sample and octadecanoic acid accounted for 40.04%. While ethyl oleate and octadecanoic acids were common to all the locations, hexadecanoic acid was present in a trace amount (1.36%) only in Kosubosu. Discussion Analysis of seed/fruit revealed that fruits from different locations are predominantly one-seeded, a few had 2 seeds per fruit, and 3 seeds per fruit are rare. Although, shea fruit is a berry, occurrence of more than two seeds in a fruit is a rare phenomenon. The higher frequency of one-seeded fruit in Kosubosu than in other locations may be due to en- vironmental and/or edaphic variations. This observation aligned with the seed number variation among fruits from the same tree as reported by Abbiw (1990) and Djekota et al. (2014) who claimed that 2–3 seeds were found in some shea Tab. 3. Phytochemical composition of shea butter from different locations in Kwara State, Nigeria. Phytochemical constituent Location Kosubosu Fufu Sare Phytate (mg g–1) 0.18 0.15 0.16 Tannins (mg g–1) 1.56 1.50 1.48 Oxalate (mg g–1) 0.80 0.82 0.78 Total phenol (mg g–1) 0.52 0.50 0.49 α-Tocopherol (mg g–1) 26.38 28.60 34.42 Alkaloid (mg g–1) 0.84 0.79 0.80 Saponin (mg g–1) 1.21 1.20 1.26 Tab. 4. Oil yield and physicochemical parameters of extracted shea butter oil from different locations of Kwara State, Nigeria. Physicochemical property Location Kosubosu Fufu Sare Oil yield (%) 54.60±2.71 46.35±2.18 49.25±2.63 Specific Gravity (g cm-3) 0.90±0.01 0.90±0.01 0.90±0.01 Acid value (mg KOH g-1) 10.58±0.70 13.56±0.94 13.16±0.91 Iodine value (g I2 (100 g)-1) 36.63±2.67 40.32±3.52 39.28±2.91 Free fatty acid (% oleic acid) 5.32±0.49 6.81±0.53 6.60±0.56 Reflective index 1.46±0.09 1.46±0.09 1.46±0.09 Saponification value (mg KOH g-1) 182.15±3.18 160.39±2.90 184.14±3.14 Peroxide value (mEq kg-1) 5.20±0.07 4.10±0.08 4.82±0.04 Viscosity 2.60±0.02 2.70±0.01 2.61±0.02 Tab. 5. Chemical composition of the extracted shea nut oil from different locations in Kwara State, Nigeria. Location Compound name Retention time (min) % composition Kosubosu Hexadecanoic acid, ethyl ester 33.175 1.36 Ethyl oleate 36.267 20.89 Octadecanoic acid, ethyl ester 36.768 15.78 2-hydroxyl-1(hydroxymethyl) ethyl ester 41.262 38.80 2-hydroxyl-1,3-propanediyl ester 41.685 23.18 Fufu Ethyl oleate 36.267 7.21 Octadecanoic acid ethyl ester 36.768 4.25 2-hydroxyl-1(hydroxymethyl) ethyl ester 44.306 47.43 Glycidol stearate 41.729 41.13 Sare Ethyl oleate 36.278 59.96 Octadecanoic acid ethyl ester 36.796 40.04 ANIMASAUN D. A., OYEDEJI S., OLORUNMAIYE K. S., AZEEZ M. A., TIJANI I. A., MORAKINYO J. A. 22 ACTA BOT. CROAT. 78 (1), 2019 fruits collected from different locations of Mondoul in the Chad Republic. Fruit and seed morphological parameters such as pericarp thickness, petiole length, seed dimensions were used to partition populations of shea tree (Djekota et al. 2014). Seed coat colours were mainly in three categories (dark brown, light brown and brown), but while light brown and brown seeds occurred frequently, only very few seeds had dark brown coat for fruits obtained from same tree in a location. Variation in seed colour observed in the pres- ent study matched previous findings on shea nut seed coat (Nafan et al. 2007, Lamien et al. 2007). Since seed formation involves meiosis, coat colouration in this case is most likely genetic and can be caused by mutation or recombination of genes. Also, these variations can be explained by natural and/ or human selection and gene flow mediated from genetic drift (Tremblay et al. 2010, Abasse et al. 2011). Differences recorded in quantitative seed characters intra and inter-lo- cational in the present study concurred with the findings on Tamarindus indica (Soloviev et al. 2004). Furthermore, sig- nificant intra and inter-locality variations have been demon- strated in shea tree and seed characters (Mbaiguinam et al. 2007, Gwali et al. 2011). The phytochemical constituents varied with locations, the variations could be accounted for by environmental stress, rainfall regimes and soil characteristics as opined by Sanou et al. (2005). Amount of phytochemicals were less than those obtained by Falana et al. (2016) who worked on V. paradoxa collected from Onipako village, Ilorin and this could be due to locational differences. The presence of simi- lar phytochemicals has been reported in other tropical plants in Nigeria and some of them exhibit varying biological ac- tivities (Sofowora 1993, Onwuliri 2004). The oil yield varied with locations, and the higher amount of oil obtained from the Kosubosu seeds could be explained by the same factors that account for the morpho- logical and biochemical compositions. The yield of the oil from the study locations were in accordance to the values earlier reported (Warra and Komo 2014). The present study revealed that in terms of oil yield, Kosubosu samples are the best. Physicochemical parameters are of great importance in determination of oil quality. Pure oils have marked ranges of specific gravity and refractive index; thus the degree of vari- ation of typical oil from its true values may indicate its rela- tive purity. The specific gravity of the extracted shea nut oil was less than 1.0 g cm–3, which implies the oil is less dense than water. The specific gravity of 0.902 g cm–3 obtained in this study correlates with the 0.90 g cm–3 documented by Raimi et al. (2014) and is congruent with those of Persea macrophylla (0.89 g cm–3) and Persea gratesima (0.90 g cm–3) (Akubugwo et al. 2008). Furthermore, the refractive index (1.468), which was the same for the locations, was also the same as the value reported by Raimi et al. (2014) and simi- lar to Anacandium occidentlis oil (1.458) (Akinhanmi et al. 2008) and walnut kernel oil (1.534) (Ozcan et al. 2010). The iodine value of the oil suggests it is a typical nondrying oil containing saturated and a low level of unsaturated fatty ac- id. Hence, the oil may be utilized for vegetable oil-based ice cream manufacturing, but, the non-drying nature of the oil makes it not applicable for paint and varnish production. Shea oil from Fufu has low saponification value compared with the other two locations. The range of saponification in- dicates that the oil may be useful in making soap and sham- poo (Ugbogu et al. 2013). Low acid value of the shea nut oil although higher than value reported by Raimi et al. (2014) was far less than of Livistona. chinensis (Nwosu et al. 2012) and this signify it could be used as an edible oil. The free fatty acid which is also an indicator of oil edibility was low and close to the range reported by Ugbogu et al. (2013). This suggests a low level of hydrolytic and lipolytic activities in the oil, thus, the extracted oil could be used as raw materi- als for industries (Obasi et al. 2012). Analysis of the fatty acid profile of shea nut oil revealed the presence of 6 fatty acids in varying quantities. It is of in- terest to note that while five fatty acids were found in Ko- subosu samples, four were present in Fufu and only two in Sare samples. Ethyl oleate and octadecanoic acids were pres- ent in all the samples irrespective of their locations. This re- sult buttresses an earlier report that variation exists in fatty acid composition of same seed oil from different locations (Wara, 2015). Ethyl oleate acid which is the most abundant in the extracted shea nut oil is an unsaturated fatty acid, natu- rally present in most seed oil and can be used for lotion and pharmaceutical solvents (PubChem, 2014). The variation in chemical constituents and fatty acid composition of the stud- ied samples may be accounted for by environmental varia- tions (Sanou et al. 2005, Mbaiguinam et al. 2007). This ex- plains why 2-hydroxyl,3-propanediyl ester, was only present in Kosubosu samples while glycidol stearate occurred only in Fufu samples. Hexadecanoic acid ethylester (palmitic ac- id), used mainly for soaps, cosmetics and release agent pro- duction, was present only in Kosubosu shea nut oil. Thus, considering the fatty acid composition of the shea nut oil, its industrial and domestic application potentials, location- al variation should be taken into consideration in collecting seeds for oil production. The present study revealed that shea tree seeds from Kosubosu had a higher yield in term of quan- tity and quality of shea nut oil than the other two locations. Conclusion The present study showed that morphological variations exist within and among locations in the seed characters of V. paradoxa from different areas of Kwara State in Central Nigeria. Also, from the results, shea nut oil phytochemical constituents and physicochemical parameters varied with lo- cations of seed collection. The fatty acid also varied with lo- cations and the presence of vital fatty acids in a large percent- age in the oil enhances the potential of shea oil as a candidate oil for industrial revolution and rural economy development. However, in sourcing for shea seed for butter and oil produc- tion, locational variations should be taken into consideration as important factors that could affect oil quantity and quality. The present study can be extended to other regions where the VARIATION IN SHEA SEEDS FROM DIFFERENT LOCATIONS ACTA BOT. 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