Synthesis, characterization, and application of olive/oleic bio-alkyd resin for determination of molybdenum in pharmaceutical samples
Elhossein A. Moawed*, Maha A. El-Hagrasy, Alaa E.A. Senan
Chemistry Department, Faculty of Science, Damietta University, Damietta, Egypt
*Corresponding author: E-mail [email protected]; Phone: 00201028921272; Fax: 0020572403868; P.O. Box: 34517- Damietta; Egypt
The study of (olive/oleic bio-alkyd resin, OOBAR) as a new biosorbent was prepared in open esterification system from an olive tree, glycerol, oleic acid and phthalic anhydride. The characterization of OOBAR was performed by using ultraviolet/visible, infrared spectra, Boehm titration, zero-point charge, iodine number, methylene blue index, density and bulk DC electrical conductivity. The data obtained show that the OOBAR surface was contained 2.6 and 1.3 mmol/g of acidic and basic sites. The pHZCP was 3.6, ?pH at pH 9.27 was -4.33, the I2 number was 2.3 mmol/g (291.9 mg/g), methylene blue was 0.69 mmol/g (220.7 mg/g), density was 0.6 g/mL and DC conductivity was 1.0 ×10-9 ?-1 cm-1. The effects of acidity, reducing agent, NH4SCN concentration, shaking time, Mo concentration, temperature, OOBAR dose, batch factor and ionic strength for the determination of Mo(V) were evaluated. The maximum OOBAR capacity for sorption of Mo(V) was 1.3 mmol/g. The molar ratio of Mo(SCN)6-, OOBAR+ was 1:1 in acidic medium. The equilibrium isotherms, kinetics and thermodynamic models for sorption of Mo(V) onto OOBAR were studied. Thermodynamic parameters such as standard enthalpy (?H), standard entropy (?S) and standard free energy (?G) was -46.2 kJ/mol, -0.082 kJ/Kmol and -21.8 kJ/mol that indicated the sorption process was spontaneous, exothermic nature with decrease disorder and randomness at the solid-liquid interface of Mo with OOBAR. Dynamic technique experiments using glass column was indicated a good affinity chromatographic separation for its applications in many pharmaceutical and biological areas including liver mice tissue and pharmaceutical vitamin drugs. The value of lower relative standard deviation (RSD%) for pharmaceutical applications samples (n=5) was found from 1.1%
Keywords: Olive; Oleic acid; Alkyd resin; Molybdenum; Pharmaceutical; Sorption
Molybdenum (Mo) has wide applications in many pharmaceutical samples 1. Mo is an economically important silvery-white transition metal and has five main oxidation states ranging from (II) to (VI). It has an atomic number of 42 and an atomic mass of 96 2,3. Mo is a bio-essential metal for humans, animals, and plants. It has relatively low toxicity because of a component or co-factor of enzymes which are important for life, so without Mo, organisms cannot function and will show signs of deficiency 4,5. Metallic Mo offers many advantages like good corrosion resistance, excellent mechanical, thermal, electrical properties, high temperature and its melting point (2883 K) because of the low coefficient of its thermal expansion and a high thermal conductivity. These excellent properties make it be widely used in electronics, metallurgy, aerospace and electrical industries 6-8.

Biosorbent is a biological origin solid system from bacterial, fungal, plant or animal origin. It has several functional groups such as carboxyl, ether, carbonyl, hydroxyl, and ester groups and more effective alternatives for metal ions sorption (e.g Mo) from aqueous solution 9-12. Adsorption using a cheap, abundant and environmental-friendly adsorbent originated from a plant such as an olive tree (Olea Europea) is currently being researched as effective substitutes as is the simplest and most useful method 13, 14. The olive tree was one of the first small fruit domesticated trees in the family Oleaceae cultivated by man for more than 5500 years ago. Its common name was for about 35 species of evergreen shrubs and trees in the olive family of the genus Olea. It has varieties of parts such as fruit, leaves, and oil. Olive trees native to the Medial East, Egypt, Palestine, Jordon, Syria, Africa and Asia. They have many varieties that exhibit major or minor phenotypical and genetic differences 15-17.

Oleic acid (OA, cis-9 C18:1) is a monounsaturated fatty acid (MUFA). It is abundant in most of the fatty acids by (48%) or in olive oil by (70%-80%). It can be extracted from peanut and its products. Researches indicate that the MUFA-rich diet has protective effects on cardiovascular risk and diabetes 18, 19.
Alkyd resins are highly branched polymer with a polyester backbone which significant for synthetic polymers 20. They are thermoplastic polyester resins can be synthesized by polycondensation 21 of three types of monomers of polyhydric alcohols (e.g; glycerol), fatty acids (e.g; oleic acid) and dibasic acids or their anhydrides (e.g; phthalic anhydride) 22-24. They can be dissolved in organic solvents and have good interactions with polar substrates such as wood and steel 25. They have excellent properties such as acceptable, inexpensive to produce, high gloss, retention color, dissolved in organic solvents, excellent thermal stability, eco-friendly, excellent wetting properties, good adhesion and good interactions with polar substrates such as wood and steel 26, 27. These properties make them suitable to apply in industrial coating, adhesives, the binder for composites and anticorrosion paint. Bio-alkyd resin contains reactive sites such as carbon-carbon double bond, a polar ester group, methylene group attached to the carbonyl site, allylic methylene attached to double bond 28.
The present paper approaches the study of OOBAR as a new biosorbent which used for Mo(V) sorption was synthesis. Properties of OOBAR are were characterized by UV-Vis, FTIR, and bulk DC electrical conductivity. The sorption of Mo(V) onto OOBAR was studied to optimize the best conditions for the sorption process. The obtained data have been examined for kinetic, thermodynamic, and equilibrium situations. Mo was applied in many pharmaceutical and biological areas including tissue of lever mice, tap water, and vitamin drugs.

2. Experimental
2. 1. Apparatus
The JASCO FTIR-4100 spectrometer in the 4000–400 cm-1 regions by using KBr disc (KBr pellet) was performed the characterization of olive bio-alkyd resin. Ammonium molybdate tetrahydrate ((NH4)6Mo7O24.4H2O) absorbance measurements were demonstrated by using JASCO (V-630 UV-VIS Spectrophotometer, Japan) at ?max 485. Bulk DC conductivity was established using Keithley, 6517B electrometer-high resistance meter after pressing OOBAR disk by a Hydraulic press at 12 tons. Dynamic technique for chromatographic separation experiments was carried out by using a glass column with a bed height of 15 cm approximately that contained 9.5 g of OOBAR was 35 cm long and 1.5 cm in diameter.
2. 2. Reagents, materials and real samples
Olive biosorbent (OB): Olive tree legs and green leaves were cut to small particle size followed by washing with water, drying at 250 ?C and blending using food blender. Carboxy olive biosorbent (COB) preparation: 100 g of OB with 80 mL of concentrated HNO3 were heating in a beaker until brown foams finished, then washing with distilled water and methanol then drier at 80 ?C.
Olive/oleic bio-alkyd resin (OOBAR) Preparation: A 85 g of oleic acid with 28 g of glycerol were refluxed for 6 h. Then the product was refluxed with 15 g biosorbent 6 h, followed by adding 15 g of phthalic anhydride then heated 6 h. The final product was washed with distilled water then methanol and left to dry at room temperature then blending and sifting at 355 µm (0.0355 cm).
Molybdenum stock solution (1 g/L): was prepared by dissolving 0.185 g of (NH4)6Mo7O24.4H2O in distilled water to 100 mL.

Vitamins drugs stock solutions: 1.217 g Vitayami for the deficiency of iron and vitamins tablets which contain (Cu: 1000 mcg; Fe: 30 mg: Mn: 2.5 mg: Mo: 15 mcg) and other content (Multi-Apex for Pharmaceutical Industries, Badr City – Cairo – Egypt), 1.067 g V2 plus multivitamins and minerals capsules which contain (Mo: 0.2 mg; Cu: 1 mg: Mn: 1 mg: Fe: 10 mg) and other content (Pharco pharmaceuticals, Alexandria – Egypt), 1.299 g Vitamax plus dietary supplements capsules which contain (Cu: 2 mg: Fe: 9 mg; Mn: 5 mg; Mo: 30 mcg) and other content (El Salam City – Cairo – Egypt) and 1.67 g Vitona plus energized and biotonic capsules which contain (Fe: 14 mg; Cu: 2 mg; Mn: 2.5 mg; Mo: 0.186 mg) and other contents (Egyptian Int. Pharmaceutical Industries CO, E. I. P. CO, 10th of Ramadan City – Egypt) were prepared by dissolving of each one in aqua regia and gently evaporated several times till dryness and removing any excess of them. The residual was dissolved in distilled water to 100 mL in a measuring flask containing 1mL of concentrated HNO3.

The tissue of lever mice stock solution: A 0.5 g of liver tissue spiked with Mo nanoparticle (0.25 mg) was prepared by dissolving in aqua regia and gently evaporated several times till dryness and removing any excess of them. The residual was dissolved in distilled water to 10 mL in a measuring flask containing 1mL of concentrated HNO3.

2.3. Recommended procedures
A 0.1 g of OOBAR was mixed with adjust concentration of molybdenum solution, H2SO4, L-ascorbic acid, and NH4SCN then diluted to 25 mL and shaken 60 min at room temperature. The remaining concentrations of Mo(V) were determined using spectrophotometrically (?max 485 nm) 29. The sorption percentage of Mo(V) and sorption capacity of OOBAR (Q, mmol/g) were calculated.
By using a dynamic technique, 10 g of OOBAR was packed through glass column which has 35 cm long and 1.5 cm in diameter with a bed height at L= 15 cm. A series of 25 mL of tap water, liver mice tissue or vitamins solutions (n = 5) were passed through the OOBAR columns at different flow rate 0.2-1.7 mL/min. The effluent solutions were collected and analyzed spectrophotometrically. Mo(V) was eluted from OOBAR columns with NH4OH (0.05 mol/L) as eluent at a flow rate of 3 mL/min then determined spectrophotometrically.

3. Result and discussion
3. 1. Characterization of olive bio-alkyd resin (OOBAR)
FTIR spectroscopy was used for identification of specific functional groups of OB, COB, OOBAR, and Mo:OOBAR in range 4000–400 cm-1. OB spectrum have broadband at 2996-3660 cm-1 (?OH), sharp peaks at 2933 cm-1 (?CH), 1612 cm-1 (?C?C) and 1084 cm-1 (?C-O-C). The bands of COB spectrum were shifted to 2343-3664, 1606 and 1097 cm-1. In addition, the new band has appeared at 37001 cm-1 while the band at 2933 was absent due to an oxidation process. Also, the bands of OOBAR spectrum were shifted to 3027-3741, 2925, 1631.5 and 1166 cm-1. The new bands have appeared at 2979, 2854 and 1739 and 1459 cm-1 due to C-H (aromatic), C-H (aliphatic), C=O and COOR. There are many sharp peaks for Mo:OOBAR was appeared at 780, 693, 519 and 507 cm-1 due to Mo(V) complexion and other bands for O-H, C-H (aromatic), C-H (aliphatic), C=O and COOR was disappeared due to the cleating agent.

Figure 1
UV-VIS electronic spectra of OB, COB, OOBAR, and Mo:OOBAR were estimated in solid state using Nujol mulls procedure. The higher energy of UV spectra bands of OB was performed at 241-265 nm which were attributed to the ?-?* transitions, and 293-340 nm which was attributed to the n-?* transitions localized on the conjugated system. Higher energy adsorption band in COB was assigned to 241-265 nm which was attributed to the ?-?* transitions, the second band was shifted to 293-355 nm and after oxidation process, there was a new band at 368-370 nm which were assigned to the n-?* transitions localized on the conjugated system. UV spectrum of OOBAR was shown that many absorption bands between 200 and 250 nm at (201, 205, 216, 224, 227-229, 232, 237, 241 and 245) nm which were assigned to ?-?* transitions and due to the several functional groups of OOBAR. Also, many lower energy bands have appeared between 300 and 320 nm due to n-?* transitions that localized on the conjugated system. Mo:OOBAR has higher energy adsorption bands at 233-245, 250 and 261 nm which were attributed to the ?-?* transitions, and 264-296 nm which was attributed to the ?-?* and n-?* transitions, and also at 299 and 307 nm which were attributed to the n-?* transitions localized on the conjugated system. There are many lower energy bands were appeared at 247, 255 and 259 nm which were assigned to ?-?* transitions, and also between 300 and 340 nm due to n-?* transitions that localized on the conjugated system.

Figure 2
Boehm titration offers an identification of the active surface sites such as carboxylic, carbonyl and phenolic. Total acidic and basic sites were detected by back titration using 0.1 mol/L HCl and NaOH solutions. Carboxylic, lactonic and phenolic groups were evaluated by using titration with 0.1 mol/L NaHCO3, Na2CO3, and NaOH solutions. The acidic and basic sites of OB were (1.8 and 0.9 mmol/g), COB was (1.6 and 0.9 mmol/g), and OOBAR was (2.6 and 1.3 mmol/g). It is clear that the amount surface of total acidic sites was greater than basic sites in OB, COB, and OOBAR which have mainly acidic character. OB has 0.7 mmol/g carboxylic sites, 0.6 mmol/g lactonic sites and 0.5 mmol/g phenolic sites. The quantity of lactonic and phenolic sites was decreased from 1.1 mmol/g in OB to 0.6 mmol/g of COB while the number of carboxylic sites of COB was increased to 1.0 mmol/g due to an oxidation process. The amount of carboxylic and carbonyl groups of OOBAR is higher than COB due to the addition of phthalic acid.

Table 1
The values of iodine number (I2 No) of OB, COB, and OOBAR were evaluated using back titration with 0.05 mol/L I2 and Na2S2O3 solutions. The I2 No of OB, COB, and OOBAR were 406.1, 320.1 and 291.9 mg/g (3.2, 2.5 and 2.3 mmol/g). The I2 No of OOBAR were decreased than OB and COB due to the coupling the biosorbent with phthalic acid. Methylene blue value (MB) of OB, COB and OOBAR were 176.97, 216.32 and 220.7 mg/g (0.59, 0.68 and 0.69 mmol/g). The increasing of MB value of OOBAR due to additional of functional groups. The results obtained indicated that the I2 No depends on the adsorption process while the MB value depends on the cation exchange process (the amount surface of total acidic sites is greater than basic sites in OB and COB). The I2 No of OOBAR was 291.9 mg/g (2.3 mmol/g) and methylene blue was 220.7 mg/g (0.69 mmol/L), these values are greater than those reported for other biosorbents 30-40.
Table 1
Table 2
The zero-point charge is the pH value that the charge of a biosorbent surface is equal to zero. The pHPZC values of OB, COB and OOBAR were7.0, 3.2 and 3.6. Maximum values of ?pH was 1.5, -1.9 at pH= 5.2, 9.1 for OB, -6.7 at pH= 11 for COB and -4.4 at pH= 9.4 for OOBAR. The surface would be negatively charged due to the deprotonation of the surface functional groups for OB, COB and OOBAR at pH values above 7.0, 3.2 and 3.6 while the surface became positively charged below pH 7.0, 3.2 and 3.6. The shifted of pHPZC value OB from pH 7 to pHs 3.2 and 3.6 for COB and OOBAR due to the increasing of functional groups in the matrix of COB and OOBAR. The OB, COB, and OOBAR sorbents have surface buffering (non-effect with acidic and basic medium) at pHs (5.4-9), (4.2-10.8), and (6.9-9.3).
The higher percentages (98%) for the sorption of molybdenum (V) from aqueous solution were in strongly acidic medium H2SO4 (0.6 – 3.6 mol/L). This was happened because of ion association complex between cationic OOBAR+ in acidic medium (pH < 3.6) and anionic ammonium thiocyanate Mo(SCN)6– complex.
Table 1
Figure 3
Figure 4
Electrical conductivity measurements were obtained by the method of Ahmenda using an EDT instrument BA380 41. Bulk electrical DC conductivity (?) was recorded at room temperature in the solid state, pressing the samples at 10 tons in the form of a circular disk and potential equal to 2 volts. The values of electrical conductivity were 0.12×10-7 and 4.1×10-6 ?-1 cm-1 for OB and COB. The value of COB was greater than OB due to increasing of active sites (functional groups) with the oxidation process. While the lower value of OOBAR than COB (0.001 ?-1 cm-1) indicates that due to converting from matrix of COB to polymer chain in OOBAR.
Table 1
3. 2. Batch sorption behavior of the sorption of Mo(SCN)6– using OOBAR
The obtained data was shown that higher values of sorption percentages were in strongly acidic medium H2SO4 (3.6 mol/L) at low concentrations of both ascorbic acid (0.08 mol/L) as reducing agent and NH4SCN (0.12 mol/L) which form orange-red color for Mo(V) complex (Fig. 4). In the effect of dose batch factor (0.05-0.5 g OOBAR in 25mL sample volume), the higher sorption percentages slightly increased (92-100%) and its capacity decreased by decreasing batch factor (v/m) and increasing amount of OOBAR dose so the sorption process can’t depend on it (Fig. 5). While in the effect of volume batch factor, the sorption percentages (98-22%) and its capacity were decreased by increasing batch factor (V/M) and sample volume (25-100 mL) (Fig. 6). By studying the effect of ionic strength of salts, the results indicate that the sorption behavior of both NaCl and KCl are similar with maximum sorption percentages (86-92%), unlike the sorption behavior percentages (80-21%) of NH4Cl indicate that the ammonium salts can be used in the stripping of Mo ion from OOBAR biosorbent (Fig. 7).
Figure 4-7
3. 3. Equilibrium studied
A 3.6 mol/L H2SO4, 0.08 mol/L ascorbic acid, 0.12 mol/L NH4SCN and shaking 1 h at room temperature, the OOBAR capacity for sorption Mo(V) was increased by increasing initial concentrations until be reached to maximum capacity (Qmax). The Qmax value of OOBAR for Mo(V) was 0.17 mmol/g (16.3 mg/g) with intercept and the correlation coefficients (R2) were 0.001 and 0.991.

Table 3
There are many isotherm models which employed to analyze the sorption mechanism and determined the parameters equilibrium. These models are Langmuir, Freundlich, Dubinin–Radushkevich, Temkin and Harkins Jura models which selected to predict the adsorption capacities and suitable for the experimental equilibrium data. Adsorption isotherm of the Langmuir model is an empirical model was obtained from an assumption of the uniform energy of adsorption sites onto the absorbent surface and the adsorbate along the plane hasn’t existed on the surface. It hypothesis that there is a fixed number of active sites is homogeneous in which adsorption occurs inside the surface of the adsorbent. These active sites have the identical affinity for a monolayer adsorption molecule and there is no interaction between molecules adsorbed. For sorption of Mo(V) complexion, the values of b, KL, and R2 were 50.5 L/mmol, 0.18 mmol/g and 0.997. Freundlich adsorption isotherm model is an empirical equation assigned to characterize adsorption on heterogeneous systems with an interaction between the adsorbed molecules, and formation of a monolayer unrestricted. It assumes due to adsorbate concentration increases; its concentration also increases on the adsorbent surface. The resulted values for sorption of Mo(V) complexion of 1/n, KF and R2 were 0.35, 0.28 L/g and 0.933. The observed results of correlation coefficients (R2) of sorption Mo(V) complexion in Langmuir and Freundlich isotherms indicated that the Langmuir model is a good fit to the adsorption experimental data and suggests monomolecular layer as well as a homogeneous distribution of active sites on OOBAR surface.

Table 4
Dubinin-Radushkevich model of adsorption isotherm is an empirical equation employed to indicate the difference between physical and chemical adsorption, describes adsorption for both heterogeneous and adsorption mechanism. R2 value was 0.975 which approximately agreement with Freundlich isotherm model, ? was -0.008 mmol2/kJ2 and KD-R was 0.19 mmol/g. The sorption free energy (?E = (-2 ?)-½) can be obtained from the value of ? as 7.91 kJ/mol.
Table 4
Temkin isotherm model was obtained as the effect of some indirect sorbate/adsorbate interactions and suggested that because of these interactions the adsorption heat of all molecules in the layer would decrease linearly. The results obtained of sorption Mo(V) complexion were R2: 0.984, A: 1.7×103 mmol/g and B: 0.03. By applied Harkins Jura adsorption isotherm model, A, B constant parameters and R2 were -1.2×10-3, 0.8 and 0.6 for sorption Mo(V) complexion.
Table 4
3. 3. Kinetic studies
Rate sorption percentage (74-91%) of Mo(V) complexion was very rapid at the initial stage of the contact period time (0.5-3 min) then rate sorption became slower until reached to the equilibrium time. This phenomenon was occurred due to the fact during the initial stage of the adsorption process, a large number of vacant surface sites were available for adsorption. Near the equilibrium, vacant surface sites remaining were difficult to occupy due to the slow Mo(V) ion pores diffusion on OOBAR and the repulsive forces between bulk phases and solid molecules.

Kinetic studies were estimated to investigate the effects of contact shaking time at a definite quantity adsorbed of initial Mo ion concentration at room temperature. It was illustrated by using five simplified kinetic models namely (pseudo-first-order and pseudo-second-order) and diffusion models (Weber and Morris intraparticle, Reichenberg, Boyd, and Bangham) to identify the rate and kinetics of sorption of Mo(V) complexion on OOBAR.

In contact shaking time batch experiments, the sorption rate of Mo(V) ion onto a given adsorbent is proportional to the adsorbed amount of Mo(V) ion from the solution phase. Adsorption kinetics can be characterized by a pseudo-first-order equation, its k1, t1/2 and R2 factors were 0.072 min-1, 9.65 min and 0.951 while resulted factors of k2, t1/2, and R2 in the pseudo-second-order kinetic model were 0.02 g/mmol min, 0.98×103 min and 1 for sorption of Mo(V) ion complex. By comparing values of correlation coefficients in both pseudo-first-order and pseudo-second-order, it clears that the pseudo-second-order is a good fit for the experiment.
Table 4
Weber-Morris or intraparticle-diffusion is a single-resistance model used to analyze the nature of the ‘rate?controlling step’. It has a basic attention due to in most liquid systems the internal diffusion determines its adsorption rate. The results obtained for R2 and ki were 0.518 and 9×10-4 mmol/g min0.5. In Reichenberg diffusion model, its linear relation Bt and t was plotted, it clears that R2 value was 0.951 for Mo(V) sorption. The better suggestion known as Boyd’s film-diffusion model is an intraparticle diffusion in a spherical particle which its Di and R2 parameters were obtained as 7.3×10-9 cm2/s and 0.518. By plotting a linear relation for Bangham’s pore diffusion model between log log Co/ (Co-Qt m) vs. log t, it does not pass through the origin and values of ko, ? and R2 were 6.3×10-5 mL/g L, 0.042 and 0.761.

Table 4
3. 4. Thermodynamic studies
Temperature solution (22-57 ?C) that effect on sorption Mo(V) complexion was studied. Sorption percentages of Mo(V) complexion were slightly decreased from 98% to 84% with increasing of temperature. Parameters value of thermodynamic obtained from batch adsorption studies were recorded by using the Van’t Hoff equation. The negative parameter values for ?G value: -21.8 kJ/mol indicated that the sorption process has a spontaneous process, ?H: -46.2 kJ/mol has exothermic nature and ?S: -0.082 kJ/K.mol at the solid-liquid interface of Mo with OOBAR has the decrease disorder and randomness.

Table 4
3. 3. Application
Application of real samples such as liver tissue mice, tap water, and vitamins drug for recovery of Mo(V) using OOBAR was estimated. A series of 25 mL from each sample were studied at optimum strong acidic medium, suitable reducing agent, NH4SCN, constant concentration and shaking for 1 hour with 0.1 g of OOBAR at room temperature.

The average recovery percentage of remaining concentration of Mo(V) was from 82-97%. The higher limit of quantitation (LOQ = 10 ?, where ? is the standard deviation) was 6-11.6 ?g/L and lower limit of detection (LOD = 3 ?) was 1.8-3.5 ?g/L which both indicates the higher sensitivity of Mo(V) determination. The addition values in liver mice tissue (0.95-0.551 mg/L) (950 – 551 ?g/L), in tap water (184-2760 ?g/L), in vitayami (50.8-33.5 ?g/L), in V2 plus (800-458.6 ?g/L) (0.8-0.46 mg/L), in vitamax plus (95.8-58.2 ?g/L) and in vitona plus (636-401 ?g/L) (0.64-0.4 mg/L) plus have recovery percentage of Mo(V) in the range 95-55% for liver mice tissue, 89-42% for tap water, 86-55% for vitayami, 100-57% for V2 plus, 80-48% for vitamax plus, 86-53% for vitona plus. Average lower values of relative standard deviation (RSD%) for real samples (n=5) were found at 1.1 % (less than 10%), which reflect the accuracy and precision of the proposed method. The result obtained was shown that OOBAR was suitable for recovery Mo(V) ions in pharmaceutical and biological samples.

Figure 8
Table 5
The sample flow rate effect depends on the recovery percentage of Mo(V) in real samples through OOBAR columns was studied. So, recovery of Mo(V) from OOBAR column were determined at different flow rates. Maximum recovery percentages (93.7-97.3%) was shown at a flow rate in the range of 0.3-1.04 mL/min in vitamins drugs. It clears that the recovery of Mo(V) from vitamins drugs was very fast and this indicated that it doesn’t depend on the change in flow rate effects. Also, at the flow rate (0.25-1.6 mL/min) and (0.2-1.2 mL/min) with recovery percentages (95-55%) and (89-42%) which decreased by increasing Mo-liver mice tissue and Mo-Tap water concentrations.

Figure 9
Table 6
The elution of Mo(V) from OOBAR columns was carried out at a flow rate of 3 mL/min by using NH4OH (0.05 mol/L) as eluent due to form ammonium molybdate complexes, then the eluent concentration was determined spectrophotometrically. The chromatogram separation of Mo(V) was completely eluted at a flow rate in the range of 0.95 to 6 mL/min with recovery percentages (32.3-0%) at first 3-15 mL in Mo-Vitamins drug.
4. Conclusion
Olive Bio-alkyd resin was successfully prepared as a new biosorbent in open esterification system. OOBAR surface was characterized by UV-VIS, FTIR, Boehm titration, zero-point charge, iodine number, methylene blue index and bulk DC electrical conductivity. From obtained thermodynamic parameters for sorption of Mo(V) which indicates a spontaneous nature for them. They indicate also chemisorption and exothermic with decreased disorder and randomness. Due to the observed correlation coefficients for Mo(V) sorption, it clears that the Pseudo-second-order is a good fit to the experimental adsorption data and also the Langmuir model is the best isotherm model which suggests monomolecular layer as well as a homogeneously. The result obtained was shown that OOBAR was suitable for recovery Mo(V) ions in pharmaceutical and biological samples including liver mice tissue and pharmaceutical vitamin drugs. The efficiency of chromatographic column decreases due to smaller plate height and larger plates number, so OOBAR column has a good effect for Mo(V) separation.

1 Bai, G. Y., Lan, X. W., Chen, G. F., Liu, X. F., Li, T. Y., & Shi, L. J. (2014). A novel sodium iodide and ammonium molybdate co-catalytic system for the efficient synthesis of 2-benzimidazoles using hydrogen peroxide under ultrasound irradiation. Ultrasonics Sonochemistry, 21, 520–526
1 Fallah, N., Taghizadeh, M., & Hassanpour, S. (2018). Selective adsorption of Mo (VI) ions from aqueous solution using a surface – grafted Mo (VI) ion imprinted polymer. Polymer, 144, 80–91.

2 Kapp, R.W. (2014). Molybdenum. Encyclopedia of Toxicology (Third Edition), 3, 383-388.

3 Arancibia, V., Romo, C. R., Aliaga, M. E., & Stegmann, E. (2018). Fast and highly sensitive method for molybdenum(VI) determination by catalytic adsorptive stripping voltammetry. Sensors and Actuators B: Chemical, 258, 612–620.
4 Burzlaff, A., Beevers, C., Pearce, H., Lloyd, M., & Klipsch, K. (2017). New studies on the in vitro genotoxicity of sodium molybdate and their impact on the overall assessment of the genotoxicity of molybdenum substances. Regulatory Toxicology and Pharmacology, 86, 279–291.

5 Wang, D. H., Sun, G. D., & Zhang, G. H. (2018). Preparation of ultrafine Mo powders via carbothermic pre-reduction of molybdenum oxide and deep reduction by hydrogen. International Journal of Refractory Metals and Hard Materials. 75, 70–77.

6 Kwon, N. Y., Kimb, Y. D., Sukc, M. J., Lee, S., & Oh, S. T. (2017). Synthesis of Mo-Si-B intermetallic compounds with continuous ?-Mo matrix by pulverization of ingot and hydrogen reduction of MoO3 powders. International Journal of Refractory Metals and Hard Materials, 65, 25–28.
7 Ku, J. G., Oh, J. M., Kwon, H., & Lim, J. W. (2017). High-temperature hydrogen-reduction process for the preparation of low-oxygen Mo powder from MoO3. International Journal of Hydrogen Energy, 42, 2139–2143.

8 Albadarin, A. B., Solomon, S., Daher, M. A., & Walker, G. (2018). Efficient removal of anionic and cationic dyes from aqueous systems using spent Yerba Mate “Ilex paraguariensis”. Journal of the Taiwan Institute of Chemical Engineers, 82, 144-155.

9 Jain, S. N., & Gogate, P. R. (2018). Efficient removal of Acid Green 25 dye from wastewater using activated Prunus Dulcis as biosorbent: Batch and column studies. Journal of Environmental Management, 210, 226 – 238.

11 Bouras, H. D., Yeddou, A. R., Bouras, N., Hellel, Nadjemi, D., Holtz, M. D., Sabaou, N., Chergui, A., Nadjemi, B. (2017). Biosorption of Congo red dye by Aspergillus carbonarius M333 and Penicillium glabrum Pg1: Kinetics, equilibrium and thermodynamic studies. Journal of the Taiwan Institute of Chemical Engineers, 80, 915-923.
12 Omar, O. S., Hussein, M. A., Hussein, B. H. M., & Mgaidi, A. (2018). Adsorption thermodynamics of cationic dyes (methylene blue and crystal violet) to a natural clay mineral from aqueous solution between 293.15 and 323.15 K. Arabian Journal of Chemistry, 11, 615-623.

13 Enenebeaku, C. K., Okorocha, N. J., Enenebeaku, U. E., & Onyeachu, B. I. (2016). Removal of Crystal Violet Dye by Adsorption onto Picrilima Nitida Stem Bark Powder: Kinetics and Isotherm Studies. IOSR Journal of Applied Chemistry, 9, 14–23.

14 Kafkaletou, M., & Tsantili, E. (2018). The paradox of oleuropein increase in harvested olives (Olea europea L.). Journal of Plant Physiology, 224–225, 132–136.

15 Baba, E., Acar, Ü., Y?lmaz, S., Zemheri, F., & Ergün, S. (2018). Dietary olive leaf (Olea europea L.) extract alters some immune gene expression levels and disease resistance to Yersinia ruckeri infection in rainbow trout Oncorhynchus mykiss. Fish & Shellfish Immunology, 79, 28-33.

16 Domínguez, H. V., Roca, M., & Rojas, B. G. (2014). Thylakoid peroxidase activity responsible for oxidized chlorophyll accumulation during ripening of olive fruits (Olea europaea L). Food Research International, 65, 247-254.

17 Souza, J. d., Preseault, C. L., & Lock A. L. (2018). Altering the ratio of dietary palmitic, stearic, oleic acids in diets with or without whole cottonseed affects nutrient digestibility, energy partitioning, and production responses of dairy cows. Journal of Dairy Science, 101, 172-185.

18 Chen, X., Li, L., Liu, X., Luo, R., Liao, G., Li, L., Liu, J., Cheng, J., Lu. Y., & Chen, Y. (2018). Oleic acid protects saturated fatty acid mediated lipotoxicity in hepatocytes and rat of non-alcoholic steatohepatitis; Life Sciences, 203, 291-304.

19 Nosal, H., Nowicki, J., Warza?a, M., Semeniuk, I., & Sabura, E. (2016). Synthesis and characterization of alkyd resins based on Camelina sativa oil, glycerol and selected epoxidized vegetable oils as functional modifiers. Progress in Organic Coatings, 101, 553–568.

20 Lawandy, S. N., Moustafa, H., Zahran, M. A. H., & Rabee, M. (2017). Effect of bio-alkyd resin oil content and viscosity on the adhesion of EPDM to polyester fabric; Journal of Adhesion Science and Technology, 32, 302–316.

21 Pierlot, C., Ontiveros, J. F., Royer, M., Catté, M., & Salager, J. L. (2018). Emulsification of viscous alkyd resin by catastrophic phase inversion with nonionic surfactant. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 536, 113-124.

22 Uzoh, C. F., Mbonu, O. F., & Onukwuli, O. D. (2016). Investigating the optimum unsaturated fatty acid content and oillength for auto-oxidative drying of palm-stearin-based alkyd resin. Progress in Organic Coatings, 101, 71 – 80.

23 Ibrahim, K. A., Abusbeih, K. A., Al-Trawneh, I., & Bourghli, L. (2014). Preparation and Characterization of Alkyd Resins of Jordan Valley Tomato Oil; Journal of Polymers and the Environment, 22, 553 – 558
24 Kurt, I., Acar, I., & Güçlü, G. (2014). Preparation and characterization of water reducible alkyd resin/colloidal silica nanocomposite coatings. Progress in Organic Coatings, 77, 949 – 956.

25 Khorasani, S. N., Ataei, S., & Neisiany, R. E. (2017). Microencapsulation of a coconut oil-based alkyd resin into poly (melamine–urea–formaldehyde) as shell for self-healing purposes. Progress in Organic Coatings, 111, 99–106
26 Gogoi, P., Boruah, M., Bora, C., & Dolui, S. K. (2014). Jatropha curcas oil based alkyd/epoxy resin/expanded graphite (EG) reinforced bio-composite: Evaluation of the thermal, mechanical and flame retardancy properties. Progress in Organic Coatings, 77, 87 – 93.

27 Lianga, L., Liua, C., Xiaoa, X., Chen, S., Huc, A., & Feng, J. (2014). Optimized synthesis and properties of surfactant-freewater-reducible acrylate-alkyd resin emulsion. Progress in Organic Coatings, 77, 1715 – 1723
28 El-Shiekh, R., Zahran, F., & Gouda, A. A. E. (2007). Spectrophotometric determination of some anti-tussive and anti-spasmodic drugs through ion-pair complex formation with thiocyanate and cobalt(II) or molybdenum(V). Spectrochimica Acta Part A, 66, 1279-1287.

29 Beltrame, K. K., Cazetta, A. L., Souza, P. S. C., Spessato, L., Silva, T. L., & Almeida, V. C. (2018). Adsorption of caffeine on mesoporous activated carbon fibers prepared from pineapple plant leaves. Ecotoxicology and Environmental Safety,147, 64–71.

29 Dai, H., Huang, Y., & Huang, H. (2018). Eco-friendly polyvinyl alcohol/carboxymethyl cellulose hydrogels reinforced with graphene oxide and bentonite for enhanced adsorption of methylene blue. Carbohydrate Polymer, 185, 1–11.
29 Soleimani, K., Tehrani, A. D., & Adeli, M. (2018). Bioconjugated graphene oxide hydrogel as an effective adsorbent for cationic dyes removal. Ecotoxicology and Environmental Safety. 147, 34–42.

30 Shaban, M., Abukhadra, M. R., Khan, A.A.P., & Jibali, B.M. (2018). Removal of Congo red, methylene blue and Cr(VI) ions from water using natural serpentine. Journal of the Taiwan Institute of Chemical Engineers, 82, 102–116.
31 Olusegun, S. J., Lima, L. S., & Mohallem, N. D. S. (2018). Enhancement of adsorption capacity of clay through spray drying and surface modification process for wastewater treatment. Chemical Engineering Journal, 334, 1719–1728.

32 Zirak, M., Abdollahiyan, A., Eftekhari-Sis, B., & Saraei, M. (2018). Carboxymethyl cellulose coated [email protected] core–shell magnetic nanoparticles for methylene blue removal: equilibrium, kinetic, and thermodynamic studies. Cellulose, 25, 503–515.

33 Wang, F. (2017). Effect of oxygen-containing functional groups on the adsorption of cationic dye by magneticgraphene nanosheets. Chemical Engineering Research and Design, 128, 155–161.

34 Gupta, N., Kushwaha, A.K., & Chattopadhyaya, M.C. (2016). Application of potato (Solanum tuberosum) plant wastes for the removal of methylene blue and malachite green dye from aqueous solution. Arabian Journal of Chemistry, 9, S707–S716.
35 Dhananasekaran, S., Palanivel, R., & Pappu, S. (2016). Adsorption of Methylene Blue, Bromophenol Blue, and Coomassie Brilliant Blue by a-chitin nanoparticles. Journal of Advanced Research, 7, 113–124.
36 Ofomaja, A. E, Pholosi, A., & Naidoo, E. B. (2015). Application of raw and modified pine biomass material for cesium removal from aqueous solution. Ecological Engineering, 82, 258–266.
37 Suneetha, M., Sundar, B. S., & Ravindhranath, K. (2015). Removal fluoride from polluted waters using low-cost active carbon derived from the bark of vitex negundo plant. Journal of analytical chemistry, 12, 33-49.

38 Mohammed, A., & Majumder, C. B. (2014). Removal of fluoride from synthetic wastewater by using bioadsorbents. Int. J. Res. Eng. Technol. 3, 776-785
39 A.B. Albadarin, S. Solomon, T.A. Kurniawan, C. Mangwandi, G. Walker, Single, simultaneous and consecutive biosorption of Cr(VI) and Orange II onto chemically modified masau stones, Journal of Environmental Management, 204, (2017), 365-374