i n d u s t r i a l c r o p s a n d p r o d u c t s 2 9 ( 2 0 0 9 ) 18

available at www.sciencedirect.com

journal homepage: www.elsevier.com/locate/indcrop

Review

Chemical composition and prole characteristics of Osage orange Maclura pomifera (Ran.) Schneider seed and seed oil
Fatnassi Saloua , Nehdi Imed Eddine, Zarrouk Hedi
Laboratory of Lipid Corps, Institute of Research and Physico-Chemical Analysis, Technical Pole, 2020 Sidi Thabet, Ariana, Tunisia

a r t i c l e
Article history:

i n f o

a b s t r a c t
Studies were conducted on the properties of seeds and oil extracted from Maclura pomifera seeds. The following values (on a dry-weight basis) were obtained for M. pomifera seed, respectively: moisture 5.88%, ash 6.72%, oil 32.75% and the high protein content 33.89%. The carbohydrate content (20.76%) can be regarded as a source of energy for animals if included in their diets. The major nutrients (mg/100 g oil) were: potassium (421.65), calcium (218.56) and magnesium (185.00). The physicochemical properties of the oil include: the saponication number 174.57; the iodine value 141.43; the p-anisidine value 1.86; the per-

Received 24 December 2007 Received in revised form 8 April 2008 Accepted 11 April 2008

Keywords: Maclura pomifera Seed oil Differential Scanning Calorimeter Physical and chemical parameters

oxide value 2.33 meq O2 /kg; the acid value 0.66; the carotenoid content 0.59 mg/100 g oil; the chlorophyll content 0.02 (mg/100 g oil) and the refractive index 1.45. Polymorphic changes were observed in thermal properties of M. pomifera seed oil. This showed absorbency in the UV-B and UV-C ranges with a potential for use as a broad spectrum UV protectant. The main fatty acids of the crude oil were linoleic (76.19%), oleic (13.87%), stearic (6.76%) and palmitic acid (2.40%). The polyunsaturated triacylglycerols (TAGs) LLL, PLL, POL + SLL, OLL, OOL (L: linoleic acid, O: oleic, P: palmitic acid and S: stearic acid) acids were the major TAGs found in M. pomifera seed oil. A relatively high level of sterols making up 852.93 mg/100 g seed oil was present. The sterol marker, -sitosterol, accounted for 81% of the total sterol content in the seed oil and is followed by campesterol (7.4%), stigmasterol (4.2%), lupeol (4.1%) and
5

-avenesterol (3.2%). The seed oil was rich in tocopherols with the following composition

(mg/100 g): -tocopherol 18.92; -tocopherol 10.80; -tocopherol 6.02 and -tocopherol 6.29. The results showed that M. pomifera seed oil could be used in cosmetic, pharmaceutical and food products. 2008 Elsevier B.V. All rights reserved.

Contents
1. 2. Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Materials and methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.1. Preparation of the material . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.2. Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.2.1. Analysis of the seed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 2 2 2 2

Corresponding author. Tel.: +216 71 537 659/666; fax: +216 71 537 688. E-mail address: fatnassisaloua@yahoo.fr (F. Saloua). 0926-6690/$ see front matter 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.indcrop.2008.04.013

i n d u s t r i a l c r o p s a n d p r o d u c t s 2 9 ( 2 0 0 9 ) 18

3.

4.

2.2.2. Physical analysis of the oil . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.2.3. Chemical analysis of the oil . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Results and discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.1. Chemical composition of M. pomifera seed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.2. Physical properties of seed oil . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.3. Chemical properties and composition of seed oil . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.3.1. Fatty acid composition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.3.2. TAGs prole . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.3.3. Tocopherols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.3.4. Sterols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

3 3 4 4 4 5 5 6 6 6 6 7

1.

Introduction

Maclura pomifera (Ran.) Schneider (SYN. Maclura aurantiaca Nutt, Ioxylon pomiferum Raf., Toxylon pomiferum Raf.ex Sarg) is a member of the Moraceae family. The plant, known as Osage orange, horse apple, mock orange or hedge apple (Wolfram et al., 1963; Wolfrom and Bhat, 1965), is actually native to southern Oklahoma and northern Texas, and is planted throughout the United States (Mahmoud, 1981). Osage orange typically occurs in open sunny areas, is very drought tolerant and can grow in a wide range of soil types and soil moisture conditions. However, it has a lower soil pH limit of 4.5. It grows best in well-drained soils. It is not only most frequently found in hedgerows or pastures, but also occurs in disturbed and oodplain forests (Sternberg, 1989). It is reported that the Osage Indians made their hunting bows from this beautiful and hard wood, and it is also used to make furniture. From April to June, Osage orange puts out its inconspicuous green owers but these are followed by the very conspicuous fruits. The fruits are 45-in. diameter, rough textured, heavy green balls which ripe to yellow green and fall in October and November. These fruits are inedible, the juice acid and milky, but squirrels relish the small seeds buried inside the pulp. When the fruits drop, they can be very messy and, for this reason, male, fruitless trees should be selected if you plant this tree. Osage orange is thorny, just like true citrus trees, and forms thickets if left to grow on its own. However, there are thornless cultivars available (Edward and Dennis, 1994). Various parts of M. pomifera are used in folkloric medicine worldwide: the decoction prepared from the roots is used for the treatment of sore eyes by the Comanche Indians in North America (Carlson and Volney, 1940), the plant sap is used in Bolivia for the treatment of tooth pain, the bark and leaves for uterine haemorrhage (Bourdy et al., 2004); and Native Americans used the Osage orange for cancer treatment (Mahmoud, 1981). Several biological activities of M. pomifera and its components including antibacterial, antifungal, antiviral, cytotoxic, antitumor, estrogenic and antimalarial activities have been reported (Peterson and Brockemeyer, 1953; Jones and Soderberg, 1979; Mahmoud, 1981; Voynova et al., 1991; Maier et al., 1995; Bunyapraphatsara et al., 2000; Hay et al., 2004). Few works have been published on the prole of M. pomifera seed oil, for example Orhan et al. (2001), but they emphasized only their fatty acid composition.

In this paper, we report the seed oil extraction from M. pomifera grown in Tunisia, in order to evaluate its physical and chemical properties, and to determine its nutritive and industrial uses.

2.

Materials and methods

This study was conducted in 2006 with M. pomifera tree from the Belvedere garden in Tunisia. The garden is located in (latitude 36 48 N; longitude 10 10 E; elevation: 3 m). The soil was characterized by medium water capacity, and low clay content. The average temperature at the time of owering until the fruit maturation was between 22 and 32 C. M. pomifera fruits were collected from plantations that were established 20 years ago and had been abandoned for several years.

2.1.

Preparation of the material

The mature fruits (harvest of November) resulting from only one tree, have approximately 0.5 g weight and 8 cm of diameter. The seeds were removed from the fruits, washed with water and then oven-dried at 60 C. Their relative percentage weight compared with the fresh fruit weight was about 11%. And prior to extraction, the seeds were ground by a grinder to pass 500 m screens.

2.2. 2.2.1.

Methods Analysis of the seed

Moisture was determined according to the AOAC Ofcial Method 930.15 (AOAC, 1999); the results are expressed in percentage. Ash and mineral contents were determined by removal carbon; about 2 g (powdered) seed, was ignited and incinerated in a mufe furnace at about 550 C for 2 h. The ask was removed from heat and left to cool. Two mililiters of H2 O2 were added and the ask was put back in a mufe furnace for further calcinations over 1 h. The total ash was expressed as percentage of dry weight. The mineral constituents (Ca, Na, K, Fe, Mg, Zn, and Cu) present in M. pomifera seed were analysed, using an atomic absorption spectrophotometer (NOUVA400, ANALYTIKJENA, Germany) and a ame ionisation spectrophotometer (Flame Photometer 410, SCHERWOOD, Germany). The phosphorus content (P) was determined by the spectrophotometric molybdovanadate method according to AOAC 970.39. Total protein was determined by the Kjeldahl method. Pro-

i n d u s t r i a l c r o p s a n d p r o d u c t s 2 9 ( 2 0 0 9 ) 18

tein was calculated using the general factor (6.25) (El-Shurafa et al., 1982). Data were expressed as percent of dry weight. Carbohydrate content was estimated by difference of mean values: 100 (sum of percentages of moisture, ash, protein and lipids) (Al-Hooti et al., 1998; Barminas et al., 1999). Oil extraction from seeds was extracted using hexane; the ground dried M. pomifera seeds (40 g) were placed into a cellulose paper cone and extracted with 400 ml hexane using a soxhlet extraction apparatus for 8 h. The solvent was removed via a rotary vacuum distillation at 4050 C ushing with nitrogen to blanket the oil during storage. The residue was weighed and stored at 20 C until it was analysed. The weight of the oil extracted was determined from 40 g of the seed powder to calculate the lipid content. The result was expressed as the lipid percentage in the seed powder dry matter.

Table 1 Norm of chemical parameters of M. pomifera seed oil Parameter

Peroxide value Para-anisidine value Acid value Iodine value Saponication value Unsaponiable matter

Norm
ISO 3960 ISO 6885 ISO 660 ISO 3961 ISO 3657 ISO 3596

and unsaponiable values were determined according to the Norm ISO (Table 1).

2.2.3.1. Fatty acid composition. The fatty acid methyl esters

(FAME) composition was determined by the conversion of oil to fatty acid methyl esters prepared by adding 1 ml of n-hexane to 40 mg of oil followed by 200 l of sodium methoxide (2 M). The mixture is heated in the bath at 50 C for few seconds followed by adding 200 l HCl (2 N). The top layer (1 l) was injected onto a GC (Agilent 6890N, California, USA) equipped with a ame ionisation detector (FID) and a polar capillary column (HP-Innowax polyethylene glycol, 0.25 mm internal diameter, 30 m in length and 0.25 m lm in thickness) to obtain individual peaks of fatty acid methyl esters. The detector temperature was 275 C and the column temperature was 150 C held for 1 min and increased at the rate of 15 C/min to 200 C and the rate of 2 C/min to 250 and held for four minute. The run time was 45 min. The fatty acid methyl esters peaks were identied comparing their retention times with individual standard FAME of lauric (C12:0), myristic (C14:0), palmitic (C16:0), palmitoleic (C16:1), stearic (C18:0), oleic (C18:1), linoleic (C18:2), linolenic (C18:3), arachidic (C20:0), eicosenoic (C20:1), behenic (C22:0) and lignoceric (C24:0) acids, approximately 99% pure, were purchased from (Supelco, USA) and analysed with the Agilent Technologies Chemstation A09.01 Software. The relative percentage of the fatty acid was calculated on the basis of the peak area of a fatty acid species to the total peak area of all the fatty acids in the oil sample.

2.2.2.

Physical analysis of the oil

The refractive index of the seed oil was determined using a Sopelem Series 3296 refractometer (Sopelem, France). The spectroscopic indices, E232 and E268 , in the UV region, were determined according to Norm ISO 3656 and the oil was diluted with isooctane. The absorbencies at 670, 610, 560, and 535 nm, related to chlorophylls and at 475, 448, and 414 nm, related to carotenoids, a 10% (v/v) solution of oil in hexane, were measured with a spectrophotometer (JASCO V-530, WITEG Labortechnik., Gmbh). The chlorophyll pigment was determined according to AOCS Ofcial Method Cc 13i-96 (AOCS, 1998), and nally the carotenoid content was determined according to AOAC ofcial method (958.05), used to evaluate the oil carotenoid content, expressed as micrograms of -carotene per gram of oil, was applied by a calibration curve constructed by preparing solutions of increasing concentration, from 0.5 to 2.5 g -carotene/ml hexane. The absorbency was recorded at 440 nm (JASCO V530, WITEG Labortechnik., Gmbh) using hexane as blank. The oil was diluted with hexane to bring the absorbance reading within the range of the calibration curve. Finally, the thermal characteristics of M. pomifera seed oil were measured by using a differential scanning calorimeter (DSC-131, SETARAM, France). A ow of nitrogen gas (1.5 ml/min) was used in the cell cooled by helium (1.5 ml/min) in a refrigerated cooling system. The instrument was calibrated for temperature and heat ow with mercury (melting point, m.p. = 38.834 C, H = 11.469 J/g), tin (m.p. = 231.928 C, H = 60.22 J/g), indium (melting point, m.p. = 156.598 C, H = 28.5 J/g) and lead (melting point, m.p. = 327.45 C, H = 24.72 J/g). The oil samples (45 mg) were weighed in open solid fat index (SFI) aluminium pans (No. S08/HBB37408, SETARAM) with an empty pan used as a reference. The sample and reference pans were then placed inside the calorimeter and kept at 70 C for 2 min. The temperature was increased from 70 to 70 C at a rate of 5 C/min. The samples were then kept at 70 C for 1 min, and then decreased again, at the same rate, up to 70 C. The scans were performed at 5 C/min.

2.2.3.2. Triacylglycerols

composition. The triacylglycerols (TAGs) prole was obtained by a reverse phase high performance liquid chromatography (HPLC) (Agilent 1100, California, USA) equipped with a G1354 quaternary pump, a G1313A standard auto sampler, and a G1362A refractive index detector. The chromatogram was carried out using Agilent Technology Chemstation software. The TAGs were separated using a commercially packed Hypersil ODS column (125 mm 4 mm) with a particle size of 3 m and were eluted from the column with a mixture of acetonitrile/acetone (65/35) at a ow rate of 0.5 ml/min; the TAG was detected with a refractive index detector. Ten microliter of 0.05 g oil diluted in 1 ml (chloroform/acetone 50/50, v/v) was injected into the HPLC. The total run time was 45 min. TAG peaks were identied by comparison with chromatograms of sunower and corn oil obtained in the same manner.

2.2.3.

Chemical analysis of the oil 2.2.3.3. Tocopherol composition. Prior to the HPLC analysis, the
seed oil 0.5 g was diluted with 5 ml hexane and 5 l sam-

The peroxide, para-anasidine acid (percentage of free fatty acid (FFA) was calculated as linoleic acid), iodine, saponication

i n d u s t r i a l c r o p s a n d p r o d u c t s 2 9 ( 2 0 0 9 ) 18

ples were injected. The tocopherol composition of M. pomifera seed oils was determined using HPLC according to norm ISO 9936. The sample was analysed by an HPLC (Agilent 1100, CA, USA) consisting of a G1354 quaternary pump, a G1313A standard auto sampler, a G1321A uorescence detector set at ( excitation = 295 nm, and emission = 330 nm) and a chemstation software. A normal phase column (Pinnacle II silica) (150 mm 3.2 mm 3 m) was used with hexane/isopropanol (99.5/0.5, v/v) as a mobile phase. The system was operated isocratically at a ow rate of 0.5 ml/min. The separations were carried out at 30 C. The quantication was based on an external standard method. The mixed tocopherol standards in a hexane solution (2 mg/ml) were prepared from the standard compounds: -, -, -, and the -tocopherols (Sigma Chemical Co., St. Louis, MO, USA).

3.
3.1.

Results and discussion

Chemical composition of M. pomifera seed

2.2.3.4. Gas chromatography analysis of phytosterols (ST). Separation of ST was performed according to the method ISO 12228. Lipids (250 mg) were reuxed for 15 min with 5 ml ethanolic KOH solution (3%, w/v) after addition of cholesterol (1 mg; Fluka) as an internal standard and a few antibumping granules. The mixture was immediately diluted with 5 ml of ethanol. The unsaponiable part was eluted over a glass column packed with slurry of aluminium oxide (Scharlau) in ethanol (1:2, w/v) with 5 ml of ethanol and 30 ml of diethyl ether at a ow rate of 2 ml/min. The extract was evaporated in a rotary evaporator at 40 C under reduced pressure, and then ether was completely evaporated under a steam of nitrogen. For the characterization of sterols, a silica gel F254 plate (Fluka) was developed in the solvent system n-hexane/diethyl ether (1:1, v/v). For the detection of sterols, the thin-layer plate was sprayed with methanol; the sterol bands were scraped from the plate and recovered by extraction with diethyl ether. The sterols trimethylsilyl ether (TMS) derivatives were prepared by adding 100 l of a silylant reagent N-methyl-N-(trimethylsilyl) triuoroacetamide/pyridine (1/10, v/v) in a capped glass vial and heated at 105 C for 15 min. Preparation of standard solutions: a mixture of standard solutions of sterols was prepared by derivatization. (cholesterol, sitosterol, stigmasterol, ergosterol and campesterol). The sterols trimethylsilyl ether derivatives were analysed using the GC system (Agilent 6890N, California, USA) equipped with a FID and the GC chemstation software. A HP-5 (5% pheynyl methyl polysiloxane column) was used (0.32 mm i.d. 30 m in length; 0.25 m lm in thickness; an Agilent 19091J-413, CA, USA). A carrier gas (helium) ow was 1.99 ml/min (splitsplitless injection with a split ratio of 1:200). The detector and the injector were set at 320 C, and the injected volume was 1 l. The total analyses were set at 71 min to ensure the elution of all ST. The operational conditions were: injector temperature 320 C, column temperature: a gradient of 4 /min from 240 to 255 C. Sterols peak identication was carried out according to the ISO 12228 reference method and conrmed by GCMS (NIST 2002 database) operating in the same conditions as used for the GCFID. All analytical determinations were performed at least in triplicate. The values of different parameters were expressed S.D.). as the mean standard deviation (x

Table 2 presents the average compositions of M. pomifera seed. The seed contained 5.88% of moisture, 6.72% ash, 20.76% carbohydrate, and crude protein and the fat contents (dryweight basis) were 33.89% and 32.75%, respectively. The yield of oil exceeded that described by (Orhan et al., 2001) for the same fruit cultivated in Turkey and that of Treculia africana which belongs to the same botanical family (Moracea) having 20.83% (Ajiwe et al., 1995). The high percentage of oil gives this seed a distinct potential for the oil industry according to Benthall (1946), Burkill (1966), and Irvine (1961). The seeds were found to be good sources of mineral elements (Table 2). The result revealed potassium to be the prevalent mineral element, followed in descending order by calcium, magnesium, phosphorus, sodium, iron, zinc and copper. The chemical composition of M. pomifera seed revealed their nutritional value for human and/or animal consumption. In order to justify the extraction of M. pomifera seed fat, it is necessary to study its functional properties.

3.2.

Physical properties of seed oil

As for the physical properties, the refractive index was studied. At room temperature, M. pomifera seed oil was present in a liquid state. The seed oil gave a refractive index reading of 1.46 (Table 2). Concerning the thermal prole, differential scanning calorimeter is a fast and direct way to assess the quality of oil (Gloria and Aguilera, 1998). Using this method, various physical properties of M. pomifera seed oil can be studied. M. pomifera seed oil exhibited a simple thermogram (Fig. 1) with two distinct peaks: a rst temperature crystallization peak having a crystallisation temperature of 32 C, crystallization enthalpy 15.58 J/g and on onset temperature 35.38 C followed by a second temperature melting peak having a melting temperature of 13.81 C, a melting enthalpy of 52.37 J/g and an onset temperature of 22.10 C. The presence of the mono and polyunsaturated fatty acids in M. pomifera seed oil (Section 3.3.1); support a higher unstable crystal form instead of most stable form. A good point concerning this oil is its ability to stay in liquid form at low temperature (13.81 C) which is useful in food application. The crude M. pomifera seed oil has showed some absorbency in the UV (100400 nm) range (Table 3). Thus, M. pomifera seed oil may be used in formulation of UV protectors against ultraviolet light UV-A and UV-B (290400 nm) responsible for must cellular damage. The high absorption at 232 nm (E232 , Table 3) may be due to the conjugation of double linkers resulting from the oxidation of polyunsaturated fatty acids and hydroperoxides of the linoleic acid resulting from the oil autoxidation. However, the low absorption at 268 nm (E268 , Table 3); indicate the absence of the secondary products of oxidation. M. pomifera seed oil contained a yellow color as indicated by the absorption between 0.42 and 0.32 (Table 3) at 414478 nm for 1% oil in hexane. These yellow colors which include carotenoids are benecial, since they simulate the appearance of butter without the use of primary colorants such as

i n d u s t r i a l c r o p s a n d p r o d u c t s 2 9 ( 2 0 0 9 ) 18

Table 2 Chemical composition, physical and chemical properties from seed and seed oil of M. pomifera Component
Chemical composition of seed Crude proteina (%) Oilb (%) Carbohydratec (%) Moisture content (%) Ashd (%) Potassiume Calciume Magnesiume Phosphoruse Sodiume Irone Zince Coppere Properties of seed oil Saponication value Iodine value (g/100 g oil) Peroxide value (meq O2 /kg) Anisidine value Unsaponiable matter (%) Index of refraction Acid value -Carotene (mg/kg oil) Chlorophyll (mg/kg oil) Physical state at room temperature
a b c d e

Table 3 Quality characteristics (color) of M. pomifera seed oil Quality characteristics

33.89 0.21 32.75 0.56 20.76 5.88 0.07 6.72 1.51 421.65 0.01 218.56 0.06 185.00 0.00 104.7 0.02 20.86 0.10 3.25 0.07 2.61 0.02 1.15 0.01 174.57 0.03 141.43 0.39 2.33 0.28 1.86 0.05 1.57 0.03 1.458 0.01 0.66 0.05 0.59 0.03 0.02 0.01 Liquid

E232 E268 A412 A414 A428 A448 A453 A475 A482 A535 A560 A610 A670 E, specic extinction; A, absorbance.

2.37 0.65 0.44 0.42 0.36 0.34 0.34 0.23 0.22 0.09 0.09 0.08 0.08

3.3.

Chemical properties and composition of seed oil

Crude protein = N (%) 6.25. Oil = weight of extracted oil 100/weight of seed. Carbohydrate obtained by difference. In %, dry matter basis. In mg/100 g of dry matter.

carotenes, annattos, and apocarotenals commonly used in the oil and fat industry (Oomah et al., 2000). The carotenoid content of M. pomifera seed oil was 5.9 mg/kg of oil (Table 2). The green pigment, particularly the chlorophyll content, usually measured at 535760 nm was negligible (0.2 mg/kg; Table 2).

Table 2 presents the chemical properties of M. pomifera seed oil. The latter shows a comparatively high iodine value (141.43) due to its high content of unsaturated fatty acids (Table 4); high iodine shows that the seed oil has the good qualities of edible oil and drying oil purposes (Eromosele et al., 1997). The acid, peroxide and para-anisidine value of M. pomifera oil showed very low values (as crude seed oil) of 0.66, 2.33 and 1.86; the peroxide values suggest that the oil has begun to degrade as expected from this highly unsaturated oil. As a result of determining the saponication number, M. pomifera seed oil showed a much lower number of (174.57) which is very low compared to safower, sunower and corn oil (OBrien, 2004) with average saponication numbers ranging between 191 and 250 (Gunstone et al., 1994). Nonetheless, the oil showed a low unsaponiable matter of 1.57%.

3.3.1.

Fatty acid composition

The most abundant fatty acids of M. pomifera oil were linoleic (76.2%), oleic (13.9%), palmitic (6.8%) and stearic (2.4%), which

Fig. 1 DSC prole of Maclura pomifera seed oil.

i n d u s t r i a l c r o p s a n d p r o d u c t s 2 9 ( 2 0 0 9 ) 18

Table 4 Fatty acid composition of Maclura pomifera seed oil (%) Fatty acid
Lauric Myristic Palmitic Palmetoleic Stearic Oleic Linoleic Linolenic Arachidic Behenic SAFA MUFA PUFA P/S

Table 6 Tocopherol and phytosterols contents of M. pomifera seed oil (mg/100 g oil) Compound
-Tocopherols -Tocopherols -Tocopherols -Tocopherols Total Campesterol Stigmasterol -Sitosterol 5 -Avenasterol Lupeol Total

Carbon length
C12:0 C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 C20:0 C22:0

Composition (%)
0.03 0.02 0.05 0.02 6.76 0.01 0.08 0.01 2.40 0.03 13.87 0.04 76.19 0.02 0.38 0.03 0.14 0.01 0.13 0.02 9.48 13.95 76.57 8.07

mg/100 g oil
18.92 0.01 6.03 0.03 6.29 0.06 10.81 0.05 42.05 63.29 0.02 36.06 0.01 691.02 0.01 27.13 0.08 35.42 0.02 852.93

SAFA, saturated fatty acids; MUFA, monounsaturated fatty acids; PUFA, polyunsaturated fatty acids.

together composed about 99.2% of the total fatty acids (Table 4). This is in agreement with the results obtained by Orhan et al. (2001). They contained a high amount of linoleic acid (76.2%), which makes them especially prone to oxidation, but which may have favorable nutritional implications and benecial physiological effects in the prevention of coronary heart disease and cancer (Oomah et al., 2000). A high ratio polyunsaturated/saturated fatty acids (8.07); is regarded favorably in the reduction of the serum cholesterol and atherosclerosis and the prevention of heart diseases (Rudel et al., 1998; Ruggeri et al., 1998). Finally, the percentage of oleic acid (13.8%) in the oil makes it desirable in terms of nutrition Corbett (2003).

(LLL) followed by palmitoyl-dilinoleoylglycerol (PLL), linoleoyloleoyl-palmitoylglycerol + dilinoleoyl-steareoylglycerol (POL + SLL), dilinoleoyl-oleoylglycerol (OLL) and linoleoyldioleoylglycerol (OOL). The amounts of monounsaturates oleoyl-dipalmitoylglycerol (PPO) were 4.32%. Good agreement between the fatty acid and triacylglycerol compositions was also found.

3.3.3.

Tocopherols

3.3.2.

TAGs prole

The triacylglycerol composition of M. pomifera showed that the majority of TAGs are in tri- and di-unsaturated form and no tri saturates were identied (Table 5). Considering the fatty acid composition; the major constituent was trilinoleoylglycerol

Having an important role for protection against oxidative deterioration of polyunsaturated fatty acids in plant material, tocopherol level in seed oils are extremely important, they are natural lipophilic antioxidants found in vegetable oils. (Table 6) shows the tocopherol content of M. pomifera seed oil: the major tocopherols were - and -tocopherol 18.92 and 10.80 mg/100 g. The -tocopherol is 45% of the total tocopherol. It is recommended for human and animal consumption because it has a higher biological activity than other tocopherols. But the -tocopherol has been suggested to possess a higher antioxidant capacity as compared to the -tocopherol. The - and -tocopherol have similar values 6.03 and 6.29 mg/100 g, respectively. The level of tocopherols in M. pomifera (43.05 mg/100 g oil) is very close to that of safower oil (3060 mg/100 g oil) (Commissione Technica, 1988) but very lower than soybean, maize and ax seed oil (Carlo et al., 2007).

Table 5 Triglycerides composition of Maclura pomifera seed oil Triglycerides

Polyunsaturated LLL OLL PLL OOL POL + SLL Monounsaturated PPO

3.3.4.

Sterols

ECN
42 44 44 46 46

Relative composition (%)

33.48 10.12 24.26 7.50 20.30 0.02 0.02 0.01 0.05 0.02

46

4.32 0.08

The levels of phytosterols (ST) in vegetable oils are used for the identication of oils, oil derivatives and for the determination of the oil quality (Artho et al., 1993; De-Blas and Del-Valle, 1996; Grob et al., 1990; Homberg, 1991; Horstmann and Montag, 1987). Furthermore, the concentration of ST has been reported to be little affected by environmental factors and/or by cultivation (Hirsinger, 1989; Homberg, 1991). Table 6 shows the distribution of the ve main phytosterols of M. pomifera seed oil: -sitosterol was the predominant component (691.02 mg/100 g) followed by campesterol (63.29), stigmasterol (36.06), lupeol (53.43), and 5 -avenesterol (27.13).

L, linoleic acid; O, oleic acid; P, palmitic acid; S, stearic acid; LLL, glycerol-trilinoleate; OLL, glycerol-oleate-dilinoleate; PLL, glycerol-palmitate-dilinoleate; OOL, glycerol-dioleate-linoleate; POL, glycerol-palmitate-oleate-linoleate; SLL, glycerol-steareatedilinoleate; PPO, glycerol-dipalmiteate-oleate. ECNs, equivalent carbon number.

4.

Conclusions

This study has revealed that M. pomifera seeds are a rich source of many important nutrients that appear to have

i n d u s t r i a l c r o p s a n d p r o d u c t s 2 9 ( 2 0 0 9 ) 18

a very positive effect on human health and give a considerable yield of oil which seems to be a good source of the essential fatty acids and the lipid-soluble bioactives. The high linoleic acid content makes the oil nutritionally valuable. The oil can protect against UV light, which justies their use in the cosmetic industry. The use of M. pomifera seed oil for industrial applications could necessitate its exposure to high thermal treatments that could lead to changes in quality characteristics of the oil. So, a study of thermo-oxidation effects on physicochemical parameters of M. pomifera seed oil must be undertaken. The production of oil from M. pomifera seed provides the use of a renewable resource, and at the same time adding value to agricultural products.

references

Ajiwe, V.I.E., Okeke, A.C., Agbo, U.H., 1995. Extraction and utilisation of breadfruit seed oil (treculia Africana). Bioresour. Technol. 53, 183184. Al-Hooti, S., Sidhu, J.S., Qabazard, H., 1998. Chemical composition of seeds date fruit cultivars of United Arab Emirates. J. Food Sci. Tech. 35, 4446. AOAC, 1999. Methods 930.15 and 958.05. In: Ofcial Methods of Analysis of the Association of Ofcial Analytical Chemists, 16th ed. Association of Ofcial Analytical Chemists, Washington, DC. AOCS, 1998. Method Cc 13i-96. In: Ofcial Methods and Recommended Practices of the American Oil Chemists Society. AOCS Press, Champaign, U.S.A. Artho, G., Grob, K., Marianai, C., 1993. On-line LCGC for the analysis of the minor components in edible oils and fatsthe direct method involving silylation. Fat. Sci. Technol. 95, 176180. Barminas, J.T., James, M.K., Abubakar, U.M., 1999. Chemical composition of seeds and oil of Xylopia aethiopica grown in Nigeria. Plant Foods Hum. Nutr. 53, 193198. Benthall, A.P., 1946. Trees of Calcutta and its neighbourhood. In: Morton J. F. (1991). The horse radish tree. A boon to arid lands. Econ. Bot. 45, 318333. Bourdy, G., de Michel, L.R.C., Roca-Coulhard, A., 2004. Pharmacopoeia in a shamanistic society: the IzocenoGuarani (Bolivian Chaco). J. Ethnopharmacol. 91, 189208. Bunyapraphatsara, N., Dechsree, S., Yoosook, C., Herunsalee, A., Panpisutchai, Y., 2000. Anti-herpes simplex virus component isolated from Maclura cochinchinensis. Phytomedicine 6, 421424. Burkill, J.H., 1966. A Dictionary of Economic Products of the Malay Peninsula, 2 Vols. Printing Works Art, Kuala Lumpur. Carlo, I.G.Tuberoso, Adam, K., Erika, S., Paolo, C., 2007. Determination of antioxidant compounds and antioxidant activity in commercial oilseeds for food use. Food Chem. 103, 14941501. Carlson, G.G., Volney, H.J., 1940. Some notes on uses of plants by the Comanche Indians. Papers of the Michigan Academy of Science. Arts Lett. 25, 517542. Commissione Technica, 1988. Olio di cartamo, caracteristiche e metodi di analisi. In: La Rivista Delle Sostanze Grasse, 65, pp. 4950. Corbett, P., 2003. It is time for an oil change! Opportunities for high oleic vegetables oils. Inform 14, 480481. De-Blas, O.J., Del-Valle, G.A., 1996. Determination of sterols by capillary column gas chromatography. Differentiation among

different types of olive oil: virgin, rened and solvent extracted. J. Am. Oil Chem. Soc. 73, 16851689. Edward, F.G., Dennis, G.W., 1994. Maclura pomifera (Osage-Orange), Fact Sheet ST-368. Institute of Food and Agricultural Sciences, University of Florida, USA. El-Shurafa, M.Y., Ahmed, H.S., Abou-Naji, S.E., 1982. Organic and inorganic constituent of dates palm pit (seeds). J. Date Palm 2, 275284. Eromosele, I.C., Eromosele, C.O., Innazo, P., Njerim, P., 1997. Short communication: studies on some seeds and seed oils. Bioresour. Technol. 64, 245247. Gloria, H., Aguilera, J.M., 1998. Assessment of the quality of heated oils by differential scanning calorimetry. J. Agric. Food Chem. 46, 13631368. Grob, K., Laufranchi, M., Mariani, C., 1990. Evaluation of olive oils through the fatty alcohols, the sterols and their esters by coupled LCGC. J. Am. Oil Chem. Soc. 67, 626634. Gunstone, F.D., John, L.H., Fred, B.P., 1994. The Lipid Handbook, 2nd ed. Chapman & Hall Chemical Database, United States. Hay, A.E., Helesbeux, J.J., Duval, O., Labaied, M., Grellier, P., Richomme, P., 2004. Antimalarial xanthones from Calophyllum caledonicum and Garcinia vieillardii. Life Sci. 75, 30773085. Hirsinger, F., 1989. New annual oil crops. In: Roebbelen, G., Downey, R.K., Ashri (Eds.), Oil Crops of the World. McGraw Hill, New York, pp. 518532. Homberg, E., 1991. Sterinanalyse als Mittel zum Nachweis von vermischungen und Verfalschungen. Fat. Sci. Technol. 93, 516517. Horstmann, P., Montag, A., 1987. Sterinanlytik zum Nachweis zu Saorol. Fat. Sci. eines Zusatzes von Sonnenblumenol Technol. 89, 381388. Irvine, F.R., 1961. Woody Plants of Ghana with Special Reference to their Uses. Oxford University Press, London. Jones, J.M., Soderberg, F., 1979. Cytotoxicity of lymphoid cells induced by Maclura pomifera (MP) lectin. Cell Immunol. 42, 319326. Mahmoud, Z., 1981. Antimicrobial component from Maclura pomifera fruit. Planta Med. 42, 299301. Maier, C.G.A., Chapman, K.D., Smith, D.W., 1995. Differential estrogenic activities of male and female plant extracts from two dioecious species. Plant Sci. 109, 3143. OBrien, R.D., 2004. Fats and oils. In: Formulating and Processing for Applications, 2nd ed. Routledge, Washington, USA. Oomah, D.B., Ladet, S., Godfrey, V.D., Liang, J., Giarard, B., 2000. Characteristics of raspberry (Rubus idaeus L.) seed oil. J. Food Chem. 69, 187193. Orhan, I., Kusmenoglu, S., Sener, B., 2001. Fatty acid composition of Maclura pomifera seed oil. Gazi Univ. Pharm. Fac. Rev. 18, 13. Peterson, C.F., Brockemeyer, E.W., 1953. The antifungal activity of an aqueous extract of osage orange wood. Am. J. Pharm. Sci. Suppl. Pub. Hlth. 125, 303310. Rudel, L.L., Kelly, K., Sawyer, J.K., Shah, R., Wilso, M.D., 1998. Dietary monounsaturated fatty acids promote aorti atherosclerosis in LDL receptor-null ApoB100-overexpressing transgenic mice. Arterioscler. Thromb. Vasc. Biol. 18, 18181827. Ruggeri, S., Cappelloni, M., Gambelli, L., Nicoli, S., Carnovale, E., 1998. Chemical composition and nutritive value of nuts grown in Italy. Ital. J. Food Sci. 10, 243252. Sternberg, Guy, 1989. Osage Orange (Maclura pomifera (Raf.) Schneider): Species Character, vol. 6. Division of Special Services, Illinois Department of Conservation, Springeld, IL, pp. 16.

i n d u s t r i a l c r o p s a n d p r o d u c t s 2 9 ( 2 0 0 9 ) 18

Voynova, E., Dimitrova, S., Naydenova, E., Karadjov, P., 1991. Inhibitory action of extracts of Maclura aurantiaca and Epilobium hirsutum on tumour models in mice. Acta Physiol. Pharmacol. Bulgaria 17, 5052. Wolfram, M.L., Komitsky Jr., F., Fraenkel, G., Looker, J.H., Dickey, E.E., McWain, P., Thompson, A., Mundell, P.M., Windrath, O.M.,

1963. Macluraxanthone and two accompanying pigments from the root bark of the osage orange. Tetrahedron Lett. 4, 749755. Wolfrom, M.L., Bhat, H.B., 1965. Osage orange pigmentsVII. 1,3,6,7-Tetrahydroxyxanthone from the heartwood. Phytochemistry 4, 765768.