Microsoft Word - 258-271 Chemistry | 240 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Simultaneous Determination of Sulfanilamide and Furosemide by Using Derivative Spectrophotometry Samar A. Darweesh Dept. of Chemistry/ College of Education for Pure Science/( Ibn Al-Haitham) / University of Baghdad. Received in:8/December/2015,Accepted in:1/March/2016 Abstract A simple, precise and accurate spectrophotometric method has been developed for simultaneous estimation of sulfanilamide and furosemide in their mixture by using first and second order derivative method in the ultraviolet region. The method depends on first and second derivative spectrophotometry, with zero-crossing and peak to base line and peak area measurements. The first derivative amplitudes at 214, 238 and 266 nm were selected for the assay of sulfanilamide and 240, 260, 284, 314 and 352 nm for furosemide. Peak area at 201- 222, 222-251 and 251-281 nm selected for estimation of sulfanilamide and at 229-249, 249- 270, 270-294, 294-333 and 333-382 nm for furosemide. The second derivative amplitudes at 220, 252 and 274 nm for sulfanilamide and 248, 272, 292, 334, and 364 nm for furosemide. Peak area at 209-229, 239-262 and 262-285 nm for sulfanilamide and at 238-253, 262-281, 281-303, 315-353 and 353-383 nm for furosemide. The first derivative absorption at 270.5 nm (zero cross point of furosemide) was used for determination of sulfanilamide and 322.5 and 352 nm (zero cross point of sulfanilamide) for determination of furosemide. The second derivative absorption at 261 and 283.3 nm (zero cross point of furosemide) was used for determination of sulfanilamide and 266, 334 and 364 nm (zero cross point of sulfanilamide) for determination of furosemide. The linearity was established over the concentration range of 1-35 μg/ml and 1-60 μg/ml for sulfanilamide and furosemide with correlation coefficient R2 0.9991 and 0.9995 respectively. Accuracy and precision of the determination method on the various amounts of sulfanilamide and furosemide with known concentrations were evaluated in their binary mixtures. The proposed method has been successfully applied to the estimation of sulfanilamide in its synthetic samples and furosemide in its drug tablets. Keywords: Derivative spectroscopy, Simultaneous determination, Sulfanilamide, Furosemide. Chemistry | 241 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Introduction Sulfanilamide chemically is 4-amino benzene sulfonamide (Figure1), it is a medicinal compound used to guard against certain bacterial infections. It is frequently used in the form of a topical cream or powder to treat surface infections, as well as a pill for internal infections. It falls into the category of sulfonamide antibacterial drugs, common infections treated by sulfanilamide include urinary tract infections, vaginal infections, strep throat, and some staph infections. Depending on the type of infection, either a cream or a pill will be prescribed [1]. Furosemide chemically is 4-chloro-N-furfuryl-5-sulfamoyl-anthranillic acid, (Figure 2) an effective diuretic, has been widely used in the treatment of chronic renal failure, hypertension, congestive heart failure and cirrhosis of the liver. Furosemide is often classified as a loop diuretic due to its predominant action in the nephron, where the drug interferes with the tubular re-absorption of sodium on Henle’s loop. Furosemide acts inhibiting the co- transportation of sodium, potassium and chloride, and further cause´s excretion of calcium, magnesium and bicarbonate ions. Intense and fast dieresis may also mask the ingestion of other doping agents by reducing their concentration in urine [2]. Various methods such as, spectrophotometric [3,4], HPLC [5-7], flow injection [8], ion- selective electrodes [9] have been reported in the literature for the determination of sulfanilamide in pharmaceutical preparations and water samples. On the other hand many publications described for the determination of furosemide include spectrophotometric [10,11], potentiometry [12,13], voltammetry [14], HPLC [15-17], GC [18], TLC [19], flow injection [20], fluorimetry [21,22] in pharmaceutical formulations and biological samples. The beginning of derivative spectrophotometry is dated on 1953 when the first analogue spectrophotometer was built by Singleton and Cooler [23]. But the fast development of this technique started in 70-s of the twentieth century, when new generation of spectrophotometers controlled by computers were constructed. An apogee of its popularity was occurred in 80-s of last century. Nowadays, it is only additional technique, rarely used, though it is fully available as a build-in function in software of modern spectrophotometers [24]. Derivative spectra can be obtained by optical, electronic, or mathematical methods. Optical and electronic techniques were used in early UV-Vis spectrophotometers, but have largely been superseded by mathematical techniques. The advantages of the mathematical techniques are that derivative spectra may be easily calculated and recalculated with different parameters, and smoothing techniques may be used to improve the signal-to-noise ratio [25,26]. A narrowing of new signals is observed during the generation of consecutive derivative spectra. This feature leads to narrowing bands and as a consequence to separate the overlapped peaks [24] and allow the assay of certain analytes from complex mixtures or matrices via mathematical interpretation of the absorption signal [27]. The purpose of the present study was to investigate the utility of derivative spectrophotometry in the assay of sulfanilamide in its synthetic samples and furosemide in its drug tablets without the necessity of sample pre-treatment. Experimental Instruments U.V.-Visible double beam spectrophotometer with 10 mm quartz cell shimadzu 1800, Windows 7 computer (DELL). Chemicals and reagents Pharmaceutical grade sulfanilamide and furosemide powder received in pure form (99.99%) were provided as a gift from the State Company for Drug Industries and Medical Appliances Samara-Iraq (SDI), sodium hydroxide (99.9 %) was provided by Panreac . All chemicals used were of analytical grade. Chemistry | 242 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Preparation of standard sulfanilamide and furosemide stock solution (100 µg.mL-1) The standard solution of sulfanilamide and furosemide were prepared by dissolving accurate weighted 10 mg of pure drug in 1-2 mL of 1 M sodium hydroxide and further diluted to 100 mL with distilled water. Preparation of synthetic sulfanilamide sample 1- To 20 mg of the bulk drug, 5 mg of interfering substance mixture (consisting of equal weights of each substance, namely, glucose, fructose, lactose, soluble starch and sucrose) were added. 2- 12.5 mg of the resulted mixture was dissolved in in 1-2 mL of 1 M sodium hydroxide and further diluted to 100 mL with distilled water to obtain 100 μg.mL-1. Preparation of furosemide from drug tablets The content of 10 tablets was grinded and mixed well. A 10 mg of the fine powder was accurately weighted to and dissolve in 1-2 mL of 1 M sodium hydroxide then diluted to 100 mL in a volumetric flask with distilled water to obtain 100 μg.mL-1. The solution was filtered by using Whatman filter paper No.41 to avoid any suspended or un-dissolved material before use, and the first portion of the filtrate was rejected. General procedures Assay procedure for the determination sulfanilamide or furosemide 1.0 mL aliquots, of sulfanilamide standard solution containing 10-350 μg (or furosemide standard solution containing 10-600 μg), were transferred into a series of 10 mL volumetric flask and diluted with distilled water. The spectrum for each solution was recorded against a distilled water as blank. Zero order spectrum was then manipulated for each to get its first derivative (D1) and second derivative (D2). Assay procedure for the determination each drug in the presence of the other 1.0 mL aliquots, of sulfanilamide standard solution containing 10-350 μg (or furosemide standard solution containing 10-600 μg), were transferred into a series of 10 mL volumetric flask containing 1.0 mL of 10 μg of furosemide solution (or 1.0 mL of 50 μg of sulfanilamide solution); the mixture was then diluted with distilled water. The spectrum for each solution was recorded against a distilled water as a blank. The recorded spectra were then manipulated to get D1 and D2. Results and discussion Absorption spectra The absorption spectra of sulfanilamide, furosemide and their mixture were recorded against distilled water as a blank. The absorption spectrum of sulfanilamide which has maximum wavelength of absorption at 252 nm, and the absorption spectrum of furosemide which appears absorption maxima at 272.5 nm, in addition to the absorption spectrum of mixture of two drugs which show a maximum wavelength of absorption at 270 nm which is related to the absorption maxima of the two compounds. Figure (3) shows the absorption spectrum of sulfanilamide and furosemide and theirs mixture. Chemistry | 243 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 First and second derivative modes Sulfanilamide and furosemide are shown broad and overlap spectrum when they are present in the same solution, therefore; they cannot determine using zero order absorption measurements. For this reason, the derivative spectroscopy method applied has the advantage that it locates hidden peak in the normal spectrum. First and second order are demonstrated to be adequate because they allow high amplitude and good spectra profile. The first and second order derivative spectra of sulfanilamide and furosemide and their mixture are shown in Figures (4 and 5) respectively. Calibration curves for sulfanilamide In order to determine the values of derivative spectra, three graphical techniques: peak to baseline (peak height), area under peak and zero crossing have been used via UV- spectrophotometric method for qualitative analyses of sulfanilamide individually and in its mixture with furosemide. The calibration curves were constructed by plotting the graphically measured (nm) amplitudes of the first and second order derivatives spectra vs. the corresponding concentrations of the examined drugs. Figure 6 shows first order spectra for sets of solutions containing various amounts of sulfanilamide (1-35 μg.mL-1) in the presence of (1 μg.mL-1) of furosemide. In the first derivative techniques, the results indicated that when the concentration of furosemide is kept constant and the concentration of sulfanilamide varied, the peak amplitudes measured at peak to baseline, zero cross, area under peak were found to be in proportion to the sulfanilamide concentration. For the second derivative technique, the spectra recorded for the previous mixtures of sulfanilamide and furosemide and the results of peak to baseline (peak height), zero cross and peak area at the specified wavelength are used to determine the exact concentration of sulfanilamide in the presence of furosemide. Figure (7) shows the measured second derivative spectra of mixture of the examined drugs. Table 1 summarizes all the results for sulfanilamide analysis by using first and second derivative technique. Calibration curves for furosemide Under the experimental conditions described, the graph obtained for UV, first and second derivative spectra showed linear relationship when calibration curves of furosemide were plotted. Peak to baseline, zero cross and area under peak techniques are also used for quantitative determination. Figure 8 shows the recorded spectra using first derivative UV spectrophotometry for solutions containing (1-60 μg.mL-1) furosemide with (5 μg.mL-1) of sulfanilamide. Mixture of furosemide (1-60 μg.mL-1) and (5 μg.mL-1) sulfanilamide of UV second derivetive spectra are obtained and presented in Figure(9). The results calculated for the determination of furosemide by proposed methods are listed in Table (2). Accuracy and precision To study the accuracy of the proposed method, the relative error percentage was carried out for five replicate analyses of two different amounts of each of the examined drug (with Beer's law). The techniques of derivative (peak to baseline, zero cross and area under the peak) were selected to deal with the recorded spectra. To determine the precision of the method, two drug solutions at studied concentration levels were analyzed each five times for both first and second order derivative spectrophotometric method, and percent coefficient of variation was calculated, Table (3) shows all results. Chemistry | 244 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Interferences study To check the interference from excipients may be used in the dosage forms, percentage recovery were calculated. This study was performed by addition of known amounts of excipients to mixture solutions of two examined drugs. First and second derivative techniques are used at the selected wavelength for the concentrations of drugs measurement. High recovery showed that no interferences were found using first and second derivative mode for the determination of sulfanilamide and furosemide in their mixture even in the presence of the excipients added, these results are listed in Table (4). Application in synthetic sample and in tablet In order to evaluate the efficiency of the derivative technique in the determination of sulfanilamide and furosemide drug, D1 and D2 procedures used for the applications of sulfanilamide in synthetic sample and furosemide in its Tablet. Good recovery % and C.V% values indicated the suitability of these methods for routine analysis of sulfanilamide and furosemide. The summary of the results is depicted in Table (5). Conclusion Derivative Spectrophotometric method was found to be rapid, simple, economical, and sensitive. It can be used in routine analysis of sulfanilamide and furosemide in their pure forms, synthetic samples and drug tablets without prior separation or treatment. Reference 1. Kent, M. (2000) “Advanced Biology”, Oxford University Press, 46. 2. Espinosa Bosch, M.; Ruiz Sánchez, A.J.; Sánchez Rojas, F. and Bosch Ojeda, C. (2013) “Analytical Determination of Furosemide: The Last Reserches” IJPBS, 3:168-181. 3. Betageri, S.; Kulkarni, M.; Shivaprasad, K. H. and Shivshankar, M. (2011) “Kinetic spectrophotometric determination of Sulfa drugs in Pharmaceutical formulations”, Der. Pharma. Chemica, 3(2):227-235. 4. Klokova, E. V. and Dmitrienko, S. G. (2008) “Spectrophotometric determination of sulfanilamides by a condensation reaction with p-dimethyl amino cinnamaldehyde” Moscow University Chemistry Bulletin 63:284-287. 5. Herrera-Herrera, A. V.; Hernandez-Borges, J.; Afonso, M. M.; Palenzuela, J. A. and Rodriguez-Delgado, M. A. (2013) “Comparison between magnetic and nonmagnetic multi-walled carbon nanotubes-dispersive solid-phase extraction combined with ultra-high performance liquid chromatography for the determination of sulfonamide antibiotics in water samples” Talanta 116:695-703. 6. Waleed, M. M.; Khaleel, N.D.H.; Hadad, G.M.; Abdel-Salam, R.A.; Haiss, A. and Kummerer, K. (2013) “Simultaneous Determination of 11 Sulfonamides by HPLC-UV and Application for Fast Screening of Their Aerobic Elimination and Bio-degradation in a Simple Test” Clean-Soil Air Water 41(9):(907-916). 7. Shaaban, H. and Go´recki, T. (2011) "Optimization and validation of a fast Ultrahigh pressure liquid chromatographic method for simultaneous determination of selected sulfonamides in water samples using a fully porous sub-2 µm column at elevated temperature " J. Sep. Sci. 35:216-224. Chemistry | 245 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 8. Icardo, M. C.; Mateo, J. V. G.; Lozano, M. F. and Calatayud, J. M. (2003) "Enhanced flow injection chemiluminometric determination of sulfonamides by on-line photochemical reaction" Analytica Chimica Acta 499:57-69. 9. Kharitonov, S. V. and Gorelov, I. P. (2000) “Ion-Selective Electrodes for Determination of Some Sulfanilamide Drugs” Pharmaceutical Chemistry Journal 34:673-676. 10. Semaana, F. S. and Cavalheiro, É. T. G. (2006) “Spectrophotometric Determination of Furosemide Based on Its Complexation with Fe(III) in Ethanolic Medium Using a Flow Injection Procedure” Analytical Letters 39:2557-2567. 11. Naveed, S.; Qamar, F. and Zainab, S. (2014) “Simple UV spectrophotometric assay of Furosemide” JIPBS 1 (3):97-101. 12. Semaan, F. S.; Pinto, E. M.; Cavalheiro, E´. T. G.; and Brettb, C. M. A. (2008) “A Graphite-Polyurethane Composite Electrode for the Analysis of Furosemide” Electroanalysis 20(21):2287-2293. 13. Santini, A. O.; Pezza, H. R.; Sequinel, R.; Rufino, J. L. and Pezza, L. (2009) “Potentiometric Sensor for Furosemide Determination in Pharmaceuticals, Urine, Blood Serum and Bovine Milk” J. Braz. Chem. Soc. 20(1):64-73. 14. Shetti, N. P.; Sampangi, L. V.; Hegde, R. N. and Nandibewoor, S. T. (2009) “Electrochemical Oxidation of Loop Diuretic Furosemide at Gold Electrode and its Analytical Applications” Int. J. Electrochem. Sci., 4:104-121. 15. Kher, G.; Ram, V.; Kher, M. and Joshi, H. (2013) “Development and Validation of a HPTLC Method for Simultaneous Determination of Furosemide and Spironolactone in Its Tablet Formulation” RJPBCS 4:365-377. 16. Rao, B. U. and Nikalje, A. P. (2012) “Determination of Furosemide and Zonisamide as a Drug Substance and in Dosage Form by Ion Pair –Reversed Phase Liquid Chromatographic Technique” Journal of Applied Pharmaceutical Science 2(5):94-99. 17. Patel, H. and Solanki, S. (2012) “Simultaneous Estemation of Furosemide and Spironolactone in Combined Pharmaceutical Dosage Form by RP-HPLC” Asian J Pharm Clin Res 5:195-198. 18. Zaporozhets, O.; Tsyrulneva, I. and Ischenko, M. (2012) “Determination of 8 Diuretics and Probenecid in Human Urine by Gas Chromatography-Mass Spectrometry: Confirmation Procedure” American Journal of Analytical Chemistry 3:320-327. 19. Wesley-Hadzija, B. and Mattocks A.M. (1982) “Thin-layer chromatographic determination of furosemide and 4-chloro-5-sulfamoyl anthranilic acid in plasma and urine” Journal of Chromatography B 229:425-432. 20. Semaan, F. S.; de Sousa, R. A. and Cavalheiro, É. T. G. (2005) “Flow Injection Spectrophotometric Determination of Furosemide in Pharmaceuticals by the Bleaching of a Permanganate Carrier Solution” J. Flow Injection Anal. 22(1):34-37. 21. Liu, Y.; Wang, H.; Wang, J. and Li, Y. (2013) “A simple and sensitive spectro-fluorimetric method for the determination of furosemide using zinc(II)- 1,4-bis(imidazol-1- ylmethyl)benzene complexes” Luminescence 28(6):882-887. 22. Semaana, F. S.; Nogueiraa, P. A. and Cavalheiro, É. T. G. (2008) “Flow Based Fluorimetric Determination of Furosemide in Pharmaceutical Formulations and Biological Samples: Use of Micelar Media to Improve Sensitivity” Analytical Letters 41:66-79. 23. Talsky, G. (1994) “Derivative Spectrophotometry” 1st edition., VCH, Weinheim. Chemistry | 246 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 24. Karpinska, J. (2012), "Basic Principles and Analytical Application of Derivative Spectro- photometry" Macro to nano spectroscopy, book edited by Jamal Uddin 253-256. 25. Owen, A. J. (1995) "Uses of Derivative Spectroscopy" UV-Visible Spectroscopy, Agilent Technologies. 26. Owen, T. (2000) "Obtaining derivative spectra" Fundamentals of modern UV-visible spectroscopy, Agilent Technologies 8. 27. Lakiss, H.; Ilie, M.; Baconi, D. L. and Bălălău, D. (2012) "Derivative UV Spectrophotometry Used for The Assay of Diazepam from Human Blood Plasma" FARMACIA 60(4):565-570. Table (1): Statistical analysis for the determination of sulfanilamide using first and second derivative spectrophotometric technique. R2 Regression equation λ (nm) Mode of calculation Order of derivative 0.9954 Y= 0.0082x+0.0012 214 Peak to base line First 0.9991 Y=0.0042x+0.0109 238 Peak to base line 0.9993 Y= 0.0042x+0.0059 266 Peak to base line 0.9999 Y= 0.0039x-0.0055 270.5 Zero cross 0.9541 Y= -0.0935x-0.4170 201-222 Peak area 0.9992 Y= 0.0771x-0.1971 222-251 Peak area 0.9990 Y= -0.0680x+0.0336 251-281 Peak area R2 Regression equation λ (nm) Mode of calculation Order of derivative 0.9995 Y= 0.0011x-0.0006 220 Peak to base line Second 0.9960 Y= 0.0005x+0.0001 252 Peak to base line 0.9991 Y=0.0003x-0.0011 274 Peak to base line 0.9963 Y= 0.0002x+0.0003 261 Zero cross 0.9988 Y=9E-05x-1E-05 283.3 Zero cross 0.9803 Y= 0.021x-0.0786 209-229 Peak area 0.9978 Y= -0.0085x+0.0111 239-262 Peak area 0.9951 Y= 0.0038x-0.0129 262-285 Peak area Chemistry | 247 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Table (2): Statistical analysis for the determination of furosemide using first and second derivative spectrophotometric technique. R2 Regression equation nm (λ) Mode of calculationOrder of derivative 0.9824 Y=0.0046x+0.023 240 Peak to base line First 0.9997 Y=0.002x-0.0075 260 Peak to base line 0.9995 Y=0.0035x+0.0167 284 Peak to base line 0.9991 Y=0.0003x+0.0014 314 Peak to base line 0.9993 Y=0.0004x-0.0001 352 Peak to base line 0.9992 Y=0.0003x+0.0007 322.5 Zero cross 0.9993 Y=0.0004x-0.0001 352 Zero cross 0.9866 Y=-0.0565x-0.287 229-249 Peak area 0.9993 Y=0.0249x-0.0872 249-270 Peak area 0.9991 Y=-0.0461x-0.1651 270-294 Peak area 0.9992 Y=0.0091x+0.1612 294-333 Peak area 0.9999 Y=-0.0119x-0.0383 333-382 Peak area R2 Regression equation nm (λ) Mode of calculationOrder of derivative 0.9982 Y=0.0006x+0.004 248 Peak to base line Second 0.9994 Y=0.0005x+0.0009 272 Peak to base line 0.9997 Y=0.0004x+0.0008 292 Peak to base line 0.9998 Y=4E-05x+0.0002 334 Peak to base line 0.9994 Y=3E-05x+0.0001 364 Peak to base line 0.9991 Y=3E-05x+8E-05 266 Zero cross 0.9998 Y=4E-05x+0.0002 334 Zero cross 0.9994 Y=3E-05x+0.0001 364 Zero cross 0.9842 Y=0.0066x+0.0126 238-253 Peak area 0.9981 Y=-0.0074x-0.002 262-281 Peak area 0.9995 Y=0.0011x+0.0293 281-303 Peak area 0.9993 Y=-0.0006x-0.0059 315-353 Peak area 0.9992 Y=0.0004x+0.0004 353-383 Peak area Chemistry | 248 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): Evaluation of accuracy and precision for the determination of sulfanilamide and furosemide by derivative technique. Drug Order of derivative Mode of analysis λ(nm) Drug conc. (μg.mL-1) RE % C.V% Taken *Found SFM First Peak to baseline 238.0 10.000 10.087 0.870 1.190 25.000 25.068 0.272 0.182 Zero cross 270.5 10.000 10.100 1.000 0.629 25.000 24.790 -0.840 0.344 Peak area 222.0-251.0 10.000 9.883 -1.170 0.486 25.000 24.873 -0.508 0.307 Second Peak to baseline 252,0 10.000 9.714 -2.860 0.559 25.000 24.969 -0.124 0.258 Zero cross 283.3 10.000 10.072 0.720 0.836 25.000 24.832 -0.672 0.475 Peak area 239.0-262.0 10.000 9.800 -2.000 0.789 25.000 25.075 0.300 0.348 Drug Order of derivative Mode of analysis λ(nm) Drug conc. (μg.mL-1) Taken *Found RE % C.V% FUM First Peak to baseline 260.0 20.000 20.072 0.360 0.201 40.000 40.060 0.150 0.078 Zero cross 352.0 20.000 19.830 -0.850 0.506 40.000 39.855 -0.363 0.127 Peak area 249.0-270.0 20.000 20.081 0.400 0.387 40.000 40.100 0.250 0.095 Second Peak to baseline 292.0 20.000 19.947 -0.260 0.603 40.000 40.084 0.210 0.219 Zero cross 334.0 20.000 20.095 0.475 0.294 40.000 40.125 0.313 0.208 Peak area 281.0-303.0 20.000 19.975 -0.120 0.437 40.000 39.920 -0.200 0.109 Chemistry | 249 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Table (4): Percent recovery for the mixtures of sulfanilamide and furosemide in the presence of 1000 μg.mL-1 of excipients. Mixture of 30 μg.mL-1 of FUM with 7 μg.mL-1 of SFM Mixture of 20 μg.mL-1 of SFM with 5 μg.mL-1 of FUM Excipients Recovery % Conc. found* of FUM (μg.mL-1) Recovery % Conc. found* of SFM (μg.mL-1) 100.67 30.200100.3720.074 Glucose 100.33 30.09899.9019.980 Fructose 98.60 29.58098.9519.790 Lactose 100.00 30.000100.4620.092 Starch 101.14 30.342100.7520.150 Sucrose *Average of three determinations. * D1 and D2 for sulfanilamide peak to baseline at 238nm and 252nm respectively. * D1 and D2 for furosemide peak to baseline at 260nm and 292nm respectively. Table (5): Application of D1 and D2 spectrophotometric techniques for the determination of sulfanilamide (taken 10 and 25 μg.mL-1) and furosemide (taken 20 and 40 μg.mL-1) in synthetic sample and in tablet. Sample Order Mode of analysis λ(nm) SFM amount (mg) Rec.% *C.V% Taken Found Synthetic sample of (SFM) First Zero cross 270.5 20.00 20.14 100.70 0.510 Second Zero cross 261.0 20.00 19.60 98.00 0.442 Zero cross 283.3 20.00 20.32 101.60 0.675 Sample Order Mode of analysis λ(nm) FUM amount (mg) Rec.% *C.V% Taken Found (FUM) in Tablet First Zero cross 322.5 40.00 39.60 99.00 0.259 Zero cross 352.0 40.00 39.75 99.83 0.319 Second Zero cross 266.0 40.00 40.50 101.25 0.504 Zero cross 334.0 40.00 39.41 98.53 0.427 Zero cross 364.0 40.00 40.63 101.58 0.362 *Average of three determinations. Figure (1): The chemical structure of sulfanilamide. Figure (2):The chemical structure of furosemide. Chemistry | 250 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): Absorption spectra of: (A) 10 μg.mL-1 sulfanilamide, (B) 20 μg.mL-1 furosemide and (C) a mixture of 10 μg.mL-1 sulfanilamide and 20 μg.mL-1 furosemide. Figure (4): First derivative spectra of (A) 10 μg.mL-1 sulfanilamide, (B) 20 μg.mL-1 furosemide and (C) a mixture of 10 μg.mL-1 sulfanilamide and 20 μg.mL-1 furosemide. Figure (5): Second derivative spectra of (A) 10 μg.mL-1 sulfanilamide, (B) 20 μg.mL-1 furosemide and (C) a mixture of 10 μg.mL-1 sulfanilamide and 20 μg.mL-1 furosemide. Chemistry | 251 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): First derivative spectra of mixture contain (1-35 μg.mL-1) sulfanilamide in the presence of (1 μg.mL-1) furosemide. Figure (7): Second derivative spectra of mixture contain (1-35 μg.mL-1) sulfanilamide in the presence of (1 μg.mL-1) furosemide. Figure (8): First derivative spectra of mixture contain (1-60 μg.mL-1) furosemide in the presence of (5 μg.mL-1) sulfanilamide. Chemistry | 252 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 Figure (9): Second derivative spectra of mixture contain (1-60 μg.mL-1) furosemide in the presence of (5 μg.mL-1) sulfanilamide. Chemistry | 253 2016) عام 2العدد ( 29مجلة إبن الهيثم للعلوم الصرفة و التطبيقية المجلد Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 29 (2) 2016 بأستعمال التقدير األني للسلفانيل أمايد والفروسيمايد مطيافية المشتقة سمر أحمد درويش جامعة بغداد / )ابن الهيثم( قسم الكيمياء/ كلية التربية للعلوم الصرفة 2016/اذار/1،قبل في:2015/كانون األول/8استلم في: الخالصة طورت طريقة طيفية بسيطة, متوافقة ودقيقة للتقدير األني لعقاري السلفانيل أمايد والفروسيمايد في مزيجهما باستعمال ى والثانية بقياسات التقاطع طريقة المشتقة االولى والثانية في المنطقة فوق البنفسجية. أعتمدت الطريقة على المشتقة االول نانومتراختيرت لتقدير 266و 238و 214الصفري وارتفاع القمة ومساحة القمة. قيم ارتفاع القمة للمشتقة االولى عند و 251-222و 222-201نانومتر للفروسيمايد. مساحة القمة عند 352و 314و 284و 260و 240السلفانيل أمايد و 382-333و 333- 294و 294-270و 270-249و 249-229اختيرت لتقدير السلفانيل أمايد وعند نانومتر 251-281 و 292و 272و 248نانومترللسلفانيل أمايد و 274و 252و 220نانومترللفروسيمايد. قيم ارتفاع القمة للمشتقة الثانية عند - 238نانومترللسلفانيل أمايد وعند 285-262و 262-239و 229-209نانومترللفروسيمايد. مساحة القمة 364و 334 270.5نانومترللفروسيمايد. امتصاص المشتقة االولى عند 383-353و 353- 315و 303- 281و 281-262و 253 نانومتر (نقطة تقاطع 352و 322.5نانومتر (نقطة تقاطع الصفر للفروسيمايد) استعملت لتقدير السلفانيل أمايد وعند نانومتر (نقطة تقاطع خ 383.3و 261امتصاص المشتقة الثانية عند الصفر للسلفانيل أمايد) استعملت لتقدير الفروسيمايد. نانومتر (نقطة تقاطع الصفر للسلفانيل 364و 334و 266الصفر للفروسيمايد) استعملت لتقدير السلفانيل أمايد وعند مايكروغرام/ مللتر 60-1مايكروغرام/مللترو 35-1الخطية المطبقة ضمن المدى لت لتقديرالفروسيمايد.أمايد) استعم على التوالي. قدرت الدقة والتوافق لطريقة التقديرعلى 0.9995و 0.9991للسلفانيل أمايد والفروسيمايد بمعامل أرتباط لتقدير لومة في المزائج الثنائية. طبقت الطريقة المقترحة بنجاحكميات مختلفة من السلفانيل أمايد والفروسيمايد بتراكيز مع اقراصه الدوائية. المصنعة والفروسيمايد في السلفانيل أمايد في نماذجه مطيافية المشتقة، التقدير األني، سلفانيل أمايد، فروسيمايد.الكلمات المفتاحية: