Microsoft Word - 245-257 Chemistry | 227 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Spectrophotometric Determination of Sulfathiazole Using 2,4 –dinitrophenylhydrazine as Coupling Reagent Israa T. Humedy Ismael M. Ali Dept. of Chemical Engineering/ College of Engineering / University of Tikrit Received in: 6/December/2015 ,Accepted in:1/March/2016 Abstract A rapid, sensitive and selective spectrophotometric method was developed for determination of sulfathiazole (STHZ) in aqueous solution. The method is based on the oxidative coupling reaction with 2,4-dinitrophenylhydrazine (2,4-DNPH) in a basic medium (pH 10.9) in the presence of potassium periodate to produce an intense orange colour, soluble in water , stable product and absorbs at 492 nm. Beer's law was in the linear range 2.0-28.0 μg/ml of sulfathiazole, the molar absorptivity, Sandellʼs sensitivity index and detection limit were 1.1437 ×104 liter. mol-1.cm-1,0.0223 μg.cm-2 and 0.1274 μg/ml respectively. The RSD value was 0.75 - 1.12 % depending on the concentration. This method was applied successfully to the determination of sulfathiazole veterinary injection liquid solution (bio prime) with average recovery of not less than 100.38 % . Keyword: Spectrophotometric, sulfathiazole, 2,4-DNPH reagent. Chemistry | 228 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Introduction The scientific name for the sulfathiazole[1] is: 4-amino-N-(thiazol-2-yl)benzenesulphonamide The molecular structure for the sulfathiazole is C9H9N3O2S2 and the molecular weight of 255.3 g/mol [2].  Sulfathiazole is an organosulfur compound used as a short-acting sulfa drug.It is used for treatment on a regular basis and prevent injuries germ and the drug used as a narcotic and prohibitive for the growth of bacteria and it was used most commonly to treat urinary areas as well as resistance to infections as sulfathiazole used as antibiotics in veterinary applications. Side effects of the drug itching ,rash, allergy, edness of skin.[3,4] Many different analytical methods were used for the determination of sulfathiazole such as spectrophotometric methods[5-7], high performance liquid chromatographic methods (HPLC)[8-14], flow-injection method[15-16], FT-Raman technique [17-18] , gas-liquid chromatographic method(GLC)[19], electrochemical method[20-21]and chemiluminescence method[22]. In this research a simple, accurate and sensitive spectrophotometric method for determining of sulfathiazole in pure form as well as in veterinary injection liquid solution (bio prime) based on the oxidative coupling using 2,4-dinitrophenylhydrazine (2,4-DNPH) in presence of potassium periodate in basic medium. Experimental Apparatus Spectrophotometric measurements have been perform using shimadzu UV-Visible spectrophotometer UV-160, ultrasonic with water bath, UNISONICS, jenway pH meter 3310, Sartorius BL210 S AG and hot plate with magnetic stirrer (BIOSAN MSH 300). Reagents and chemicals used All chemicals and analytical reagents used in this research are of high purity. Preparation of solutions 1-Standard sulfathiazole solution,1000 μg/ml-1 The solution was prepared by dissolving 0.1 g of sulfathiazole in amount of distilled water and the volume is diluted to 100 ml with distilled water in a volumetric flask. 20 ml of this solution diluted to 100 ml with distilled water, to obtain a solution with a concentration of 200 μg/ml (7.834× 10-4 M).This solution was prepared to be used not more than one month. The absorption spectrum of this solution versus distilled water on Figure (1) shows that the a maximum absorption of this solution is 286 nm. 2- 2,4-dinitro phenylhydrazine reagent solution (2 ×10-3M) The solution was prepared by dissolving 0.1981g of 2,4-dinitro phenyl hydrazine in 5 ml of sulphuric acid and the volume is completed to 100 ml in a volumetric flask with distilled water, then 20 ml of this solution diluted to 100 ml with distilled water to obtain a solution with a concentration of (2 ×10-3M). Chemistry | 229 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 3-Potassium periodate solution (5×10-3M ) A 0.1150g of potassium periodate was dissolved in amount of distilled water using ultrasonic and the volume is completed to 100 ml in a volumetric flask with distilled water. -4 Interference solutions 1000 µg / ml A 0.1000 g of each foreign compounds was dissolved in distilled water then the volume is completed to 100 ml in a volumetric flask with distilled water. 5-Sodium hydroxide solution, ( approximate concentration 1.0 M) The solution was prepared by dissolving 4.0 g of sodium hydroxide in 100 ml of distilled water in a volumetric flask, and the solutions for the lowest concentration are prepared with dilution. 6- Solution of STHZ injection formulation (1000 µg/ml) Veterinary injection liquid solution (bio prime) (Bioagripharm GmbH-germany), every 1.0 ml contains 40 mg of sulfathiazole, the solution was prepared as follows: The solution was prepared by taking an equivalent of 0.100 g from sulfathiazole and the volume has been completed to 100 ml with distilled water to obtain a solution with a concentration of 1000 µg/ml. A solution of 200 µg/ml is prepared by dilution of 20 ml of the above solution by distilled water in a volumetric flask of 100 ml. Preliminary investigations A 1 ml of 2,4- DNPH reagent is added to 1.5 ml of standard STHZ solution in the presence of 1 ml of potassium periodate solution in basic medium using 0.5 ml of 1.0 M sodium hydroxide then diluted with distilled water in a 25 ml volumetric flask to produce an orange color product. Absorption spectrum of the colored dye against its corresponding blank reagents shows maximum absorption at 492 nm in contrast to blank reagent. Results and discussion Optimization of the experimental conditions The effect of various variables on the absorption intensity of 1.5 ml of sulfathiazole solution (200µg/ml), 1.0 ml of (2,4-DNPH) and 1.0 ml of KIO4 in alkaline medium(0.5 ml,1.0M NaOH ) was studied to establish the optimum conditions. Selection of the oxidizing agent The study was conducted by adding 1.0 ml of different types of oxidizing agents (5×10-3M) to 1.0 ml of 2,4-dinitro phenyl hydrazine solution (2×10-3M) and 0.5 ml of sodium hydroxide solution(1.0 M)such as: ammonium ceric sulphate dihydrate,  N-bromosuccinimde  potassium hexacynoferrate(III), potassium periodate, potassium iodate and ferric sulphate, the results showed the potassium periodate solution gives a higher intensity for colored product at 492 nm compared with other oxidizing agents used , so this oxidizing agent was selected in subsequent experiments. Effect of pH The effect of pH was studied by adding 0.1-2.5 ml of 1.0 M sodium hydroxide solution. The best pH is found to be in the range of 9.8–11.2, so the pH of 10.9 and 1.5 ml of sodium hydroxide solution was adopted in subsequent experiments, the results are shown in Table (1).  Chemistry | 230 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Effect of the amount of oxidizing agent The study was conducted to select the best amount of oxidizing agent KIO4 (5×10-3M) by adding different volumes (0.3-2.5 ml) of oxidizing agent to volumetric flasks containing 1.5 ml of STHZ (200 µg/ml) and 1.0 ml of 2,4-DNPH  reagent (2×10-3M), then addition of 1.5 ml of 1.0 M sodium hydroxide and the volume was completed to 25ml with distilled water. The results shown in the Table (2) indicate that the volume of 1.2 ml of oxidizing agent KIO4 (5×10-3M) is the optimum amount because of highest absorbance, so it was selected in subsequent experiments. Effect of the amount of coupling reagent The effect of the amount of coupling reagent was studied by adding different volumes (0.3- 2.5ml) of reagent solution (2×10-3 M) to the volumetric flasks containing 1.5 ml of sulfathiazole ( 200µg/ml) and 1.2 ml of oxidizing agent KIO4 (5×10-3M), then the addition of 1.5 ml of 1.0 M sodium hydroxide and the volume is completed to 25ml with distilled water, the results are shown in Table (3), it is clear that the volume of 2.0 ml of 2,4-DNPH reagent (2×10-3M) is the optimum amount because it gave the highest absorption. So it is used in subsequent experiments. Order of additions The effect of different orders of addition on the absorption of the colored product was studied.The results are shown in Table (4) indicate that the addition (STHZ 2,4−DNPH KIO4 OH-) achieves a higher absorption of colored product. So it was adopted in subsequent experiments. Effect of oxidation time The color intensity reached maximum, after drug it reacted with 2,4-DNPH and KIO4 for 10 min in basic medium, therefore, a 10 min is sufficient for the oxidation to be completed, so it is adopted in the subsequent experiments. The color obtained was stable for 70 min and the results are shown in Table (5). Effect of temperature The effect of temperature (5-60°C) on the absorption of the colored product was studied.  The results are shown on Figure (2) indicate that the optimum temperature is (25°C)  because it gave the best absorption. So it is used in subsequent experiments. Effect of the solvents The effect of the solvents on the formed colored product was studied, the dilution was carried out by different organic solvents instead of water. The results are shown in Table (6) indicate that the water is a good medium for reaction and gives good absorption value at the wavelength of 492 nm and due to its availability, it has been used as the best solvent in the subsequent experiments. Final absorption spectra The spectrum of the colored product by coupling of 1.5 ml of sulfathiazole solution (200µg/ml) with 2.0 ml of 2,4-DNPH (2×10-3M) in the presence of 1.2 ml of KIO4 (5×10-3M) in basic medium (1.5 ml,1M NaOH ) (pH 10.9 ) and temperature 25°C against its corresponding reagent blank show a maximum absorption at 492 nm in contrast to the blank reagent. The spectra are shown on Figure (3). Chemistry | 231 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Procedure for construction of calibration curve To a series of volumetric flasks (25ml), 0.25-3.5ml of (200 µg/ml) of sulfathiazole were transferred, 1.2ml of KIO4 (5×10-3 M) and 2.0 ml of 2,4-DNPH reagent (2×10-3M), 1.5 ml of 1.0M sodium hydroxide solution(pH 10.9 )were added at 25°C . After that the solutions were left for 10 min to complete the reaction, then the volumes were completed to the mark with distilled water. The absorbance was measured at 492 nm against the blank reagent. Figure (4) illustrates that the calibration curve is linear over the concentration range of 2.0 -28.0 μg/ml while higher concentrations show a negative deviation from Beer's law. The molar absorptivity value is 1.1437 × 104 liter. mol-1.cm-1and the Sandellʼs sensitivity index 0.0223µg/cm2. Accuracy and precision Accuracy and precision were studied by measuring absorption at 492 nm for two different concentrations of the drug within the limits of Beer's law, the average recovery (99.88 %) and the relative standard deviation ( 1.12 %) indicate that the method is of high accuracy and precision . The results are shown in Table (7). Detection limit Detection limit was calculated by measuring the absorption for the lower concentration 2 µg/ml at optimal conditions at 492 nm. The results are shown in Table (8). The nature of the formed product To know the nature of the formed orange color complex (stoichiometry of drug with the reagent), Job's method and molar ratio method were applied. In both methods, the concentration of each of the standard STHZ solution and 2,4-DNPH reagent solution is equal to 7.834× 10-4 M. In Job's method, in a series of volumetric flasks (25 ml ), different volumes of the drug solution ranging from 1-9 ml and different volumes (9-1 ml) of reagent solution were mixed. A 1.2 ml of potassium periodate (5×10-3 M) and 1.5 ml 1 M of sodium hydroxide solution were added and volumes were completed to the mark with distilled water. The absorbance was measured at 492 nm against the blank reagent. The results as it Figure (5) show that the ratio is 1:1. In molar ratio method, 1.5 ml of the standard drug solution (7.834× 10-4 M ) in a series of volumetric flasks (25 ml) were transferred and different volumes 0.25 – 4.0 ml of 2,4-DNPH reagent solution, 1.2 ml of potassium periodate (5×10-3M) and 1.5 ml 1.0 M of sodium hydroxide solution were added. The volumes were completed to the mark with distilled water and the absorbance was measured at 492 nm against the blank reagent. Molar ratio was found to be 1:1. The results are shown in Figure (6) which is in agreement with the Job's method results . Scheme 1 shows the formed complex structure. Calculation of the stability constant Stability constant for the dye formed was calculated according to mole ratio method under the optimum conditions. For the reaction between a drug and reagent giving ML complex, degree of dissociation and stability constant was calculated [23, 24]: M + L ML K = – If the α degree of dissociation and C concentration of colored product,so: K = – – α – Am = is the greatest value of the absorption As = absorption value at the equivalence point (when the ratio of product 1:1) Chemistry | 232 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 The stability constant was calculated for the value of AS is taken (0.458) when the volume of both (drug and reagent) is 1.5 ml of 7.834× 10-4 M, while the value of Am (0.482) is taken at volume of 4 ml for 7.834× 10-4 M reagent. The value of the stability constant, is found to be 4.89×105 liter.mol-1 for colored product STHZ - 2,4-DNPH under optimal experimental conditions and this indicates that the product is of high stability. Effect of interferences In order to test the efficiency and selectivity of the proposed method, the effect of some foreign substances that usually present in dosage forms was studied by taking volumetric flasks ( 25 ml) containing 1.5 ml of sulfathiazole (200 µg / ml) , then different volumes (2.5 , 5 ,7.5 ml) of foreign substances (1000 µg / ml) were added resulting in a final concentration of (100, 200 ,300 µg / ml ). The optimum conditions were applied and the volumes were completed to the mark with distilled water.The absorbance was measured at 492 nm versus blank reagent and recovery were calculated. The results showed that there is no interferences ,Tablet (9). The proposed equation for reaction can be written as follows: Scheme (1) Applications This method was applied for the determination of STHZ in its pharmaceutical formulation bio prime injection (40 mg ). Direct method In this method, different volumes (0.25, 0.5ml) of a pharmaceutical formulation solutions (200 µg/ml) were transferred to 25 ml volumetric flasks and the resulting concentrations (2, 4 µg/ml) were treated as in construction of calibration curve. The absorbance was measured at 492 nm for six times. Recovery and RSD were calculated and Table (10) shows the efficiency and success of the developed method for the determination of STHZ in its pharmaceutical formulation, the average recovery is 100.38 %. Standard additions method To prove that the developed method is free from interferences , method of standard additions is applied for determining of STHZ in its pharmaceuticals. Different volumes (0.25, 0.5ml) of a pharmaceutical formulation solutions ( 200 µg/ml) were transferred to six volumetric flasks ( 25 ml) for each volume, then increasing volumes (0.25-2.0 ml) of 200 µg/ml of STHZ standard solution were added with leaving the sixth flask without addition. The solution was treated as in construction of calibration curve. The absorbances were measured at 492 nm (Figure 7) the measured concentration was calculated from the NO2 O2N NHNH2 NO2 O2N N KIO4  NaOH N N O2N NO2 Orange dye NO2 O2N NN N NH2 S O O N H N S Chemistry | 233 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 equation of the straight line and the results shown in Table (11) indicate that method of standard additions is in consistent with the direct method within the acceptable range of error, indicating that the method is satisfactory and free from interferences. The average recovery is 101.13%. Conclusions The results obtained confirm that the proposed method is simple, rapid and of good sensitivity for the determination of sulfathiazole. The method is based on oxidative coupling between sulfathiazole and 2,4-dinitrophenylhydrazine reagent in presence of potassium periodate in basic medium to form orange colored dye which is water soluble, stable and shows a maximum absorption at 492 nm. This method does not require temperature control, nor use of organic solvents, or solvent extraction and it can be applied successfully for determination of sulfathiazole in veterinary pharmaceuticals formulation with recovery of not less than 100. 38%. References 1-British Pharmacopeia, (2005). 5th Ed., by system simulation ltd., the stationary office, London, in CD-ROM" 2-British Pharmacopeia,(2009). 6th Ed., by system simulation ltd., the stationary office, London, in CD-ROM" 3- Harvey, R. A.; Clark, M. A.; Finkel, R. ; Rey, J . A. and Whalen, K.; (2012). 5th Ed.; "Lippincotts Illustrated Reviews : Pharmacology". 4- www. webmd.com 5- Azeez, Y. J. (2009) ; "Spectrophotometric determination of sulfathiazole in different pharmaceutical formulations", Wasit Journal for Science & Medicine 2(1), 30 - 38. 6-Dombrowski, L. J.; Browning, R. S. and Pratt, E. L. (1977) ; "Direct spectrophotometric determination of sulfathiazole in presence of sulfadiazine and sulfamerazine", J. Pharm. Sci., 66(10),1413-1415. 7- Salinas,F.; Espinosa Mansilla, A. and Berzas Nevado, J.J.(1991); "Simultaneous determination of sulfathiazole and oxytetracycline in honey by derivative spectrophotometry", Microchemical Journal , 43(3), 244–252. 8- Zheng, H.; Wang, P. and Li, J.(2007) ; "Determination of 12 sulfonamides in cosmetics by ultra- performance liquid chromatography", Se. Pu., 25(2),238-240. 9- Wu, Y.; Zhao,L.; Liu, Y.; Jiang,Y.; Liu, X. and Shen, J.(2007) ; "Simultaneous determination of nine sulfonamide residues in milk using solid phase extraction and high performance liquid chromatography", Se. Pu., 25(5), 728-731. 10- Martel, A. C. and Zeggane, S.(2003) ; "HPLC determination of sulfathiazole in French honeys", Journal of Liquid Chromatography & Related Technologies, 26(6), 953- 961. 11- Albert, K.; Riter, K.L. and Smallidge, R.L.(2003) ; " Determination of sulfathiazole in type C medicated swine feed by reversed-phase liquid chromatography with post- column derivatization", J. AOAC. Int.,86(4),623-630. 12- Smallidge, R.L.; Kentzer, E.J.; Stringham, K.R.; Kim, E.H.; Lehe, C.; Stringham, R.W. and Mundell, E.C. (1988) ; "Sulfamethazine and sulfathiazole determination at residue levels in swine feeds by reverse-phase liquid chromatography with post- column derivatization", J. Assoc. Anal. Chem.,71(4),710-717. 13- Clark,S.B.; Turnipseed, S.B.; Madson,M.R.; Hurlbut, J.A.; Kuck, L.R. and Sofos, J.N. (2005) ; "Confirmation of sulfamethazine, sulfathiazole, and sulfadimethoxine residues in condensed milk and soft-cheese products by liquid chromatography/tandem mass spectrometry", J. AOAC. Int., 88(3), 736-743. Chemistry | 234 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 14- Fang, G.Z.; He, J.X. and Wang, S. (2006) ; "Multiwalled carbon nanotubes as sorbent for on- line coupling of solid-phase extraction to high-performance liquid chromatography for simultaneous determination of 10 sulfonamides in eggs and pork", Journal of Chromatography A, 1127(1–2),12–17. 15- Gui-Hua, L.I.U; De-Man, H.A.N. ; Hua-Ding, L.I.A.N.G. ; Shou- Wan, T.A.N.G. ; Fu- You P.A.N. and Rui-Qiang, Y.A.N.(2012); "Determination of Four Kinds of ulfonamides in Aquatic Products by Flow Injection On-line Pre concentration andHigh Performance Liquid Chromatography", Chinese Journal of Analytical Chemistry, 03. 16- Evgen'ev, M. I. ;  Garmonov, S.Yu. and Shakirova, L. Sh. (2002) ; "Flow-Injection determination of sulfanilamides in drugs and biological fluids with spectrophotometric detection", Journal of Analytical Chemistry, 57(1), 64-70. 17- Sanchez, M. L. ; Rama, M.J. R. ; Medina, A. R.; Diaz , A. M. and Canada, M. J. A.(2008) ; "Pharmaceutical powders analysis using FT-Raman spectrometry: simultaneous determination of sulfathiazole and sulfanilamide", Talanta, 74(5),1603-1607. 18- Rama, M.J. R. ; Sanchez, M. L.; Medina, A. R.; Diaz , A. M. and Canada, M.J. A. (2005);"Flow- through sensor with Fourier transform Raman detection for determination of sulfonamides", Analyst, 130(12),1617-1623. 19- Munns, R.K. and Roybal, J.E.(1983) ; " Rapid gas-liquid chromatographic method for determination of sulfathiazole in swine feed", J. Assoc. Off Anal. Chem., 66(2),287-290. 20- Giahi, M. ; Pournaghdy, M. and Rakhshaee, R.(2009) ; "A new lidocaine-selective membrane electrode based on its sulfathiazole ion-pair"; Journal of Analytical Chemistry, 64(2),195-200. 21- Rizzotto, S. B. y. M. ; Okulik, N. and Jubert, A. (2007) ; "The interaction between sulfathazole and cobalt(II): potentiometric studies"; Quim. Nova,30(5), 1136-1142. 22- Liu, J. ; Fang, G. ; Zhang, Y. ; Zheng, W. and Wang, S.(2009); "Development of a chemiluminescent enzyme-linked immunosorbent assay for five sulfonamide residues in chicken muscle and pig muscle"; Journal of the Science of Food and Agriculture, 89(1), 80–87. 23- Bosque-Sendra, J.M.; Almansa-Lopez, E.; Garcia-Campana, A.M. and Cuadros, L. (2003);"Rodriguez; Data analysis in the determination of stoichiometries and stability constants of complexes"; Anal. Sci., 19,1431-1436. 24- Charan, D.D. (2011) ; "Analytical Chemistry"; PHI Learning Pvt. Ltd., 79-85. Table (1 ) Effect of base pH Absorbance ml of 1.0 M NaOH 5.4 0.198 0.3 7.9 0.251 0.5 8.6 0.278 0.7 9.8 0.322 1.0 10.4 0.358 1.2 10.9 0.377 1.5 11.2 0.345 2.0 11.6 0.294 2.5 Table (2) Effect of the amount of oxidizing agent. 2.5 2.0 1.5 1.2 1.0 0.7 0.5 0.3 ml of KIO4 (5×10-2M) 0.355 0.361 0.384 0.421 0.378 0.349 0.287 0.218 Absorbance Chemistry | 235 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Table (3) Effect of the amount of coupling reagent. 2.5 2.0 1.5 1.2 1.0 0.7 0.5 0.3 ml of 2,4-DNPH (2x10-3M) 0.463 0.489 0.454 0.438 0. 422 0.395 0.331 0.271 Absorbance Table (4) Order of additions. Order number Order of addition Absorbance I STHZ 2,4−DNPH KIO4 OH- 0.489 II -OH 2,4−DNPH4KIO STHZ 0.475 III -OH +STHZ4KIO DNPH-2,4 0.156 IV DNPH-2,44KIO −OH STHZ 0.108 V STHZ +4KIO −OH DNPH-2,4 0.242 Table (5) Effect of oxidation time. Time(min) 5 10 15 20 25 30 35 40 50 60 Absorbance 0.267 0.490 0.489 0.489 0.490 0.488 0.488 0.487 0.485 0.485 Table ( 6 ) Effect of the solvents. Propanol Isobutanol Aceton Methanol Ethanol Water Solvent 0.284 0.173 0.335 -------- 0.264 0.489 Absorbance 490 503 500 turbid 485 492 λmax, nm Table (7) Results of accuracy and precision. RE% Average recovery% Recovery*% RSD% Conc. of STHZ(ppm) -0.53 99.88 99.47 1.12 4 0.29 100.29 0.75 8 * Average of six determinations Table (8) Detection limit. D.L µg/ ml S x (Absorption) Concentration µg/ ml 0.1274 0.00153 0.075 2 * Average of ten determinations Table ( 9 ) Effect of interferences. Recovery (%) of 12 µg . ml-1 of STHZ per µg . ml-1 foreign compound added Foriegn compound 300 200100 100.15 98.76 99.53 Starch 99.85 99.35100.25Glucose 96.95 98.13 100.29 Fructose 98.18 97.5497.39Maltose 102.02 100.39 100.66 Sucrose Table (10) Direct method for determination of STHZ in bio prime injection. Recovery*,% RSD,% RE,% STHZ measured µg/ml STHZ present µg/ml 101.50 1.03 1.50 2.03 2 99.25 1.36 -0.75 3.97 4 * Average of six determinations Chemistry | 236 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Table (11) Results of standard additions method. Figure (1) :absorption spectrum of the sulfathiazole versus distilled water Figure (2): Effect of temperature on the absorption of the colored complex 0 0.1 0.2 0.3 0.4 0.5 0.6 0 10 20 30 40 50 60 70 A b so rb a n ce Temp  ͦC Recovery, (%) RE% STHZ measurd µg/ml STHZ present µg/ml Type of Drug 102.50 2.52.052 Injection .99 75 -0.25 3.99 4 A b so rb a n ce Wavelength,(nm) 0.50 Chemistry | 237 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Figure (3): Final absorption spectrum of the colored complex (from 200 μg/ml Sulfathiazole). SB : Absorption spectrum of colored complex versus blank. SW: Absorption spectrum of colored complex versus distilled water. BW: Absorption of blank versus distilled water. Figure (4): Calibration curve for determination STHZ by oxidative coupling with 2,4-DNPH reagent. Figure (5): Job's method of formed product by oxidative coupling of STHZ(7.834× 10-4M) with 2,4-DNPH reagent (7.834× 10-4 M). 0 0.1 0.2 0.3 0.4 0.5 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 A b so rb a n ce [STHZ] /[STHZ] + [2,4‐DNPH] 0.50 SW BW SB A b so rb an ce Wavelength, (nm) Chemistry | 238 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Figure (6): Molar ratio method of formed product by oxidative coupling of STHZ(7.834× 10-4 M) with 2,4-DNPH reagent(7.834× 10-4 M). Figure (7): Standard additions curve for the determination of STHZ in injection. y = 0.045x + 0.0922 R² = 0.9979 y = 0.0449x + 0.1793 R² = 0.9981 ‐0.2 0 0.2 0.4 0.6 0.8 1 ‐6 ‐4 ‐2 0 2 4 6 8 10 12 14 16 18 A b so rb a n ce Concentration of STHZ, µg/ml 0.25 ml, 2 ug/ml 0.5 ml, 4 ug/ml 0 0.1 0.2 0.3 0.4 0.5 0.6 0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 A b so rb a n ce ml of Reagent Chemistry | 239 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 كاشف االقتران عمالباست التقدير الطيفي للسلفاثايازول هيدرازين نيتروفنيلثنائي -4,2 حميدياسراء طالب اسماعيل محسن علي قسم الهندسة الكيمياوية/كلية الهندسة/جامعة تكريت 2016/اذار/1،قبل في: 2015/كانون األول/6استلم في: الخالصة هذه .تستند المائي المحلول في السلفاثايازول عقار لتقدير وحساسة وانتقائية سريعة طيفية طريقةيتضمن البحث تطوير بوجود العامل المؤكسد في وسط قاعديثنائي نيتروفنيل هيدرازين -4,2الكاشف مع التأكسدي االقتران تفاعل الطريقة على نانوميتر. 492الموجي الطول عند امتصاص أعلى ويعطي الماء في ذائب برتقالي اللون ناتج لتكوينالبوتاسيوم بيريودات 1.1437االمتصاصية الموالرية و. السلفاثايازولايكروغرام/مل من م 28.0 - 2.0 كانت حدود قانون بير في مدى التراكيز - 0.75االنحراف القياسي النسبي ت قيمة. وتراوح2-سم.مايكروغرام 2230.0وداللة ساندل 1- .سم 1- لتر.مول 410 × الحقن سائل محلول في لتقدير السلفاثايازول بنجاح الطريقة هذه تطبيق تمّ . مايكروغرام/مل 0.1274، وحد كشف % 1.12 .% 100.38من اقل ليست وبمعدل استرجاعية )بايوبرايم(البيطري ثنائي نيتروفنيل هيدرازين. - 4,2الطريقة الطيفية, السلفاثايازول, :الكلمات المفتاحية