Actually, Glimepiride (GLI) and Timolol (TIM) are analysed by several analytical techniques such as HPLC, Uv Vis and electrochemical methods, but in this study these compounds were determined by use Britton-Robinson (B-R) buffer pH3 in mobile phases as a new work. B-R pH3 : Acetonitrile : Water : Methanol (10 : 30 : 10 : 50 % v/v/v/v) and B-R pH3 : Acetonitrile : Methanol (30 : 15 : 55 % v/v/v), were used as mobile phases for chromatographic studies of GLI and TIM respectively. 1´10-3 mol L-1 of GLI and TIM were prepared  by dissolving the suitable weight of analyte in methanol and distilled water respectively, in 50 ml volumetric flask. GLI and TIM were HPLC separated from interferences and analytical determined using C-18 (5µm) with UV detector at 254 nm and 1.00 mL min-1 flow rate by 10 µL injection volume. The calibration curves for GLI and TIM  were studied over the range 1×10-4 to 1×10-3 mol L-1 (correlation coefficient = 0.994, n=6) for GLI and (correlation coefficient = 0.996, n=6) for TIM. The HPLC reproducibility and stability were evaluated for 5×10-4 mol L-1 of GLI and TIM , yielded good results for ten HPLC measurements with relative standard deviations RSD% were 0.43 and 1.1%, and good stability observed. The developed HPLC was applied to determine of GLI and TIM in pharmaceutical formulations with mean recoveries 103% ±0.71 and 99% ±1.0, respectively.   

F. Alghamdi, A.

S. Alkhalaf, Abdulhameed

S. Alkhalaf, Abdulmajeed

English

Revues Scientifiques Marocaines

HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC DETERMINATION OF GLIMEPIRIDE AND TIMOLOL IN PHARMACEUTICAL FORMULATIONS USING BRITTON -ROBINSON BUFFER IN MOBILE PHASES

 Ali F. Alghamdi & al. / Mor. J. Chem. 4 N°2 (2016) 454-560  454      HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC DETERMINATION OF GLIMEPIRIDE AND TIMOLOL IN PHARMACEUTICAL FORMULATIONS USING BRITTON -ROBINSON BUFFER IN MOBILE PHASES  Ali F. Alghamdi*, Abdulmajeed S. Alkhalaf, Abdulhameed S. Alkhalaf    Department of Chemistry, Faculty of Science - Taibah University, P.O Box 30002, Al-Madinah Al- Munawarah-Saudi Arabia   *Corresponding author: alifh2006@hotmail.com  Received 01 April 2016, Revised 04 May 2016, Accepted 11 May 2016  Abstract  Usually, the analytical techniques used to analyze Glimepiride (GLI) and Timolol (TIM) are HPLC, UV-Vis and electrochemical methods. In this study, we developed a new procedure for its determination by using Britton-Robinson (B-R) buffer pH3 in mobile phases. B-R pH3 : Acetonitrile : Water : Methanol (10 : 30 : 10 : 50 % v/v/v/v) and B-R pH3 : Acetonitrile : Methanol (30 : 15 : 55 % v/v/v), were used as mobile phases for chromatographic studies of GLI and TIM respectively. 1.010-3 mol L-1 of GLI and TIM were prepared by dissolving the suitable weight of analyte in methanol and distilled water respectively, in 50 ml volumetric flask. GLI and TIM were HPLC separated from interferences and analytically determined using C-18 (5µm) with UV detector at 254 nm and 1.00 mL min-1 flow rate by 10µL injection volume. The calibration curves for GLI and TIM were studied over the range 1.0×10-4 to 1.0×10-3 mol L-1 (correlation coefficient = 0.994, n=6) for GLI and (correlation coefficient = 0.996, n=6) for TIM. The HPLC reproducibility and stability were evaluated for 5.0×10-4 mol L-1 of GLI and TIM, yielded good results for eight measurements with relative standard deviations RSD% were 0.44 and 1.43%, and good stability is observed. The developed HPLC is applied in order to determine of GLI and TIM in pharmaceutical formulations with mean recoveries 103% ±0.71 and 99% ±1.00, respectively.  Keywords:  HPLC, Glimepiride, Timolol, mobile phase, Britton-Robinson, pharmaceutical formulations.  1. Introduction Glimepiride (GLI) is chemically identified as 1-[[p-[2-(3-ethyl-4-methyl-2-oxo-3-pyrroline-1carboxamido) ethyl]phenyl] sulfonyl]-3-(trans-4-methylcyclohexyl) urea (C24H34N4O5S) with a molecular weight of 490.62 (see scheme 1) and named commercially "AMARYL". Glimepiride is a white crystalline powder and is practically insoluble in water but it is forthwith soluble in methanol. Furthermore, Glimepiride is used for controlling blood sugar in people, Whereas, The management of blood sugar with glimepiride can decrease  Ali F. Alghamdi & al. / Mor. J. Chem. 4 N°2 (2016) 454-560  455   the risk of eye, kidney and nerve damage. On the other hand, Timolol (TIM) is a non selective beta adrenergic receptor blocking agent. Its chemical IUPAC name is (-)-l-(tert-butyl amino)-3- [(4-morpholino-l, 2, 5-thiadiazol-3-yl)oxy] propan-2-ol. Timolol has a molecular weight of 316.421 (see scheme 2). It is a white crystalline powder that is soluble in water, methanol and ethanol. Timolol is used to treat open angle glaucoma and other causes of high pressure inside the eye. High-performance liquid chromatography (HPLC) is a famous technique to usually determine amino and nucleic acids, proteins, hydrocarbons, carbohydrates, drugs, terpenoids, pesticides, antibiotics, steroids, metal organic species and a variety of inorganic substances [1]. There are several analytical methods used to analysis Glimepiride such as: chromatographic methods [2-11], spectrophotometry [12-14] and electrochemical techniques[15-18]. Timolol also analytical determined by many techniques such as: chromatographic methods [19-26], spectrophotometry [27-29] and electrochemical techniques [30,31]. To the best of our knowledge, the determination GLI and TIM using Britton-Robinson (B-R) buffer in mobile phases is new and not published anywhere else. The precision, reproducibility, affectivity and sensitivity of HPLC were also evaluated in this study through determinations of GLI and TIM in pharmaceutical formulations.  NONHOSOONHNHO Scheme 1. Chemical structure of Glimepiride (GLI)  ONNSNO NOHCH3H3CCH3H  Scheme 2. Chemical structure of Timolol (TIM)  2. Experimental 2.1. Apparatus               An ultimate 3000, thermo scientific DIONEX (US made) instrument was used for  high performance liquid chromatographic measurements, with UV-Vis detector, 20 µL manual loop injector and C18 column 5 µm, connected computer dell OPTIPLEX 3010 (China made). The chromatograms were printed via a hp laser jet pro 200 color M251n, Hewlett-Packard development company, (China made). Oxford adjustable micropipette (Ireland made) was used to inject micro-liter volumes of GLI and TIM standard solutions and pharmaceutical samples. Drops of 1.00 mol L-1 of HCl and NaOH were used to control pH values in the used B-R buffers by HANNA INSTRUMENT pH 211 (Romanian Made). Micropipettes 100-1000 µL OXFORD (Ireland made ) were used to inject the standard solutions and analyzed samples.   Ali F. Alghamdi & al. / Mor. J. Chem. 4 N°2 (2016) 454-560  456   2.2. Materials Stock solutions of Glimepiride (GLI) and Timolol (TIM) and commercial samples were prepared in different concentrations to be suited for HPLC procedures. Methanol and Acetonitrile HPLC-grade 0.2 micron filtered, Fisher scientific, product of Trinidad, New Jersey, were used in HPLC mobile phase. In addition, Britton-Robinson (B-R) buffer was prepared by use boric, acetic and phosphoric acids [32] in pH 3 for injection in mobile phases. Methanol and distilled water solvents were used for solubility of GLI and TIM compounds, respectively.   2.3. Chromatographic conditions The HPLC instrumental parameters are: 1.00 ml/min flow rate, 30oC column temperature, UV detector; 254 nm wavelength, 20µL injection volume and C-18 (5µm) HPLC column.  HPLC retention times were recorded of 3.40 and 3.46 min for GLI and TIM respectively. A 10 µL of studied solutions was injected in the HPLC column over chromatographic measurements. The used mobile phases contained: B-R pH3 : Acetonitrile : Water : Methanol (10 : 30 : 10 : 50 % v/v/v/v) and B-R pH3 : Acetonitrile : Methanol (30 : 15 : 55 % v/v/v) for GLI and TIM, respectively.                                                                                           2.4. GLI and TIM preparations 1.010-3 mol L-1 stock solutions of GLI and TIM (manufactured by QASSIM PHARMACEUTICAL PLANT BURAYDAH, SPIMACO ADDWAEIH, BURAYDAH, KSA) were prepared by dissolving the appropriate amounts of GLI and TIM standards in methanol and distilled water solvents in 50 ml volumetric flasks. Then GLI and TIM solutions should be stored in laboratory under room temperature. The needed concentrations of GLI and TIM determinations were prepared by diluting their stock solutions by solvents.                                2.5. Commercial drugs preparations  The content of GLI is commercially available as Glimephan-2 tablet; 2mg GLI (ACINO, MANUFACTURED BY ACTAVIS LTD, MALTA FOR ACINO, LIES BERG, BASEL- LAND, Swiss made), while the content of TIM is commercially available as eye drops, OCUMOL (timolol 0.5%, 5mg/mL), PHARMA-SA, RIYADH. These commercial drugs were obtained from the local pharmacy in MADINA-KSA. 20 tablets of GLI contain 40 mg of GLI, which were grinded, weighted, and dissolved in methanol solvent then 1.010-3 mol L-1 GLI was prepared in 50 ml volumetric flask. A 1.010-3 mol L-1 of TIM was directly calculated and prepared from eye drop in 25 ml volumetric flask. The HPLC method was applied for the determination of commercial samples; GLI after filtration steps and TIM at room temperature.                                                                                                                                3. HPLC Results and Discussion 3.1. Eexperimental and Optimization parameters In general, the mobile phase, flow rate, wavelength, detector, column temperature and retention time are very important parameters for chromatographic measurements. In this study, 5.010-4 mol L-1 of GLI and TIM were studied by HPLC technique where it was given high and sharp signals at mobile phases: B-R pH3 : Acetonitrile : Water : Methanol (10 : 30 : 10 : 50 % v/v/v/v) and B-R pH3 : Acetonitrile : Methanol (30 : 15 : 55 % v/v/v) for GLI and TIM, respectively, 1ml min-1 flow rate, 254 nm wavelength, UV detector, 30oC  Ali F. Alghamdi & al. / Mor. J. Chem. 4 N°2 (2016) 454-560  457   column temperature. Under optimum conditions, HPLC signals were obtained and recorded for 5.010-4 mol L-1 of GLI and TIM at 3.40 min and 3.46 min retention times as shown in figures 1 and 2.   Figure 1. HPLC signal of 5.010-4 mol L-1 GLI at 3.40 min retention time and B-R pH3: Acetonitrile: Water: Methanol (10 : 30 : 10 : 50 % v/v/v/v) mobile phase; (Abs= 156)   Figure 2. HPLC signal of 5.010-4 mol L-1 TIM at 3.46 min retention time and B-R pH3: Acetonitrile : Methanol (30 : 15 : 55 % v/v/v) mobile phase; (Abs= 45)  3.2 Analytical performance of HPLC  3.2.1 Calibration graphs Under optimum HPLC parameters, excellent linear relationships were obtained between HPLC signals and concentrations of GLI and TIM over the range 1.010-4 – 1.010-3 mol L-1 as illustrated in figures 3 and 4. The least-square equation was used to calculate the calibration curve parameters for GLI and TIM as shown in the form; where Abs is absorbance value, C is molar concentration, r2 is correlation coefficient and n is number of HPLC measurements. Abs =  2.12 105 C  + 22                    r2 = 0.994            n = 6;    (GLI) Abs =  4.14 104 C  + 2.6                    r2 = 0.996            n = 6;    (TIM)   Ali F. Alghamdi & al. / Mor. J. Chem. 4 N°2 (2016) 454-560  458    Figure 3. Calibration graph of GLI  at 3.4 min retention time and B-R pH3: Acetonitrile : Water : Methanol (10 : 30 : 10 : 50 % v/v/v/v) mobile phase   Figure 4. Calibration graph of TIM  at 3.462 min retention time and B-R pH3: Acetonitrile : Methanol (30 : 15 : 55 % v/v/v) mobile phase    3.2.2 Reproducibility and Stability of HPLC Signals  The reproducibility was considered as an important parameter to evaluate the developed HPLC precision. It was measured eight times for 5.010-4 mol L-1 of GLI and TIM, yielded, relative standard deviation (RSD) which calculated to be become 0.44% and 1.43% for GLI and TIM respectively as reported in table 1. On the other hand, The HPLC signals of 5.010-4 mol L-1 of GLI and TIM were monitored and rerun every 10 min for 1.5 hours. There were good stabilities for HPLC measurements of GLI and TIM.          Ali F. Alghamdi & al. / Mor. J. Chem. 4 N°2 (2016) 454-560  459   Table 1. HPLC Reproducibility for eight measurements of 5.010-4 mol L-1 of GLI and TIM  GLI and TIM Molarity concentration GLI Absorbance (mAU) TIM Absorbance (mAU) Average ± S.D. GLI and TIM RSD % GLI and TIM    5.010-4  mol L-1  155.0 23.5    155.9±0.68 & 23.3±0.33    0.44% & 1.43 % 155.0 23.0 156.0 23.0 156.0 23.0 156.0 23.0 156.0 23.8 157.0 23.5 156.5 23.6  4. Applications  The developed HPLC technique was used for analytical application of GLI and TIM in pharmaceutical samples. GLI is commercially available as tablets which were determined by HPLC after dissolving and filtration processes. Twenty tablets which contained 40 mg GLI, were dissolved in methanol as solvent and filtrated to be determined by HPLC method. While TIM eye drop was injected directly in HPLC column. These measurements were given results for GLI and TIM as can be recorded in table 2.   Table 2. HPLC results of GLI and TIM in pharmaceutical formulations   Labeled Contents GLI  tablet 2.0 mg & Eye drop 0.5 % TIM GLI Founded (mg)  Recovery % TIM Founded (%)  Recovery % Average ± S.D. GLI Average ± S.D. TIM 2.08 104 0.500 100   103% ±0.71    99%±1.00   2.06 103 0.500 100 2.04 102 0.490 98 2.06 103 0.495 99 2.06 103 0.495 99  5. Conclusion The GLI and TIM have been determined in commercially pharmaceutical drugs, for the first time, by using a developed HPLC technique in the presence of B-R buffer in the used mobile phase under selection parameters which given high and sharp HPLC signals. Calibration graph, reproducibility and stability were evaluated for analytical performance of HPLC method. GLI and TIM drugs obtained from the local pharmacies in ALMADINA- KSA.   Acknowledgement The authors would like to thank Mr. AlQarni and Mr. AlRibdy in QASSIM PHARMACEUTICAL PLANT BURAYDAH, SPIMACO ADDWAEIH, SAUDI ARABIA for their assistances during this article.     Ali F. Alghamdi & al. / Mor. J. Chem. 4 N°2 (2016) 454-560  460   References [1] D. A. Skoog, J. J. Leary, Principles of instrumental analysis, 4rd,  Saunders college publishing USA, 1992. [2] S. R. Polagani., N. R. Pilli, R. Gajula R., V. Gandu, J. Pharm. Anal., 3 (2013) 9–19.  [3] I. I. Salem, J. Idrees, J. I. Tamimi, J. Chromatograph. B Analyt Technol. Biomed. Life Sci., 799 (2004) 103-109. [4] K. Hohyun , K. Y. Chang, H. P. Chang, M. S. Jang, J. Lee, H. J. Lee, K. R. Lee, Chromatographia, 60 (2004) 93-98.  [5] K. H. Hyun, L. H. Joo, K. L. Chang, K. Y. Han, S. Beom, Bull. Korean Chem. Soci., 25 (2004) 109-114.  [6] K. S. Lakshmi, T. Rajesh, S. Shrinivas, Inter. J. Pharm. Tech. Res., 1 (2009) 494-499. [7] S. Sujatha, R. T. Sandhya, V. Ciddi, Amer. J. Anal. Chem., 2 (2011) 152-157.  [8] I. U. Khan , F. Aslam, M. Ashfaq, M. N. Asghar, J. Anal. Chem., 64 (2009) 171-175. [9] L. Rabbaa‐Khabbaz, R. Abi Daoud, D. Karam‐Sarkis, A. Chawki, A. Zoghbiet, J. Liq. Chromatograph. Rel. Techno., 28 (2005) 3255-3263. [10]  N. M. El-Enany, A. A. Abdelal, F. B. Fathalla, I. Yoshinori,  M. N. Nakamura, Chem. Cent. J., 6 (2012) 9. [11] S. Alok, A. P. Tripathi, A. V. Dewani, Eur. J. Anal. Chem., 11 (2016)55-61.  [12] B. Rudy, B. A. Magali, R. N. Hérida, J. Braz. Chem. Soc., 22 (2011) 292-299. [13] S. Bhargavi, S. Gopisetty, K. S. Dantu, K. Ashok, S. Nama, Int. J. Pharm. Sci. Rev. Res., 21 (2013) 131-133. [14] N. A. Alarfaj, E. A. Al-Abdulkareem, F. A. Aly, Asia.  J. Chem., 23 (2011)3441-3444. [15] H. L. Ma, M. T. Xu, P. Qu, X. H. Ma, Acta. Pharmaceutica. Sinica, 40 (2005) 750-753.   [16] S. Incilay, A. Sacide,  Cur. Anal. Chem., 12 (2016) 333 – 340.  [17] W. A. Badawy, M. A. El-Ries, I. M. Mahdi, Talanta, 82 (2010)106-112. [18] A. M. Radi, S. H. Eissa, Collect. Czech. Chem. Commun., 76 (2011) 13-25. [19] S. P. Kulkarni, P. D. Amin, J. Pharm. Biomed. Anal., 23 (2000) 983-987.  [20] C. Boiero, D. Allemandi, M. Longhi, J. M. Llabot, J. Pharm. Biomed. Anal., 111 (2015) 186-189.  [21] N. Erk, Pharmazie, 58 (2003) 491-493.   [22] R. V. Rele, V. V. Mhatre, J. M. Parab, C. B. Warkar, J. Chem. Pharm. Res., 3 (2011)138-144. [23] W. A. Shadoul, E. A. Gad Kariem, M. E. Adam, K.E.E. Ibrahim, Elixir. Pharma., 38 (2011) 4060-4063. [24] A. Phogat, M. S. Kumar, N. Mahadeva, Inter. J. Rec. Adv. Pharm. Res., 3 (2011) 31-36. [25] R. D. Marini, P. Chiap, B. Boulanger,  S. Rudaz, E. Rozet, J. Crommen, P. Hubert, Talanta, 68 (2006) 1166–1175.   [26] J. Mehta, V. Patel, N. Kshatri, V. Niranjan, Anal. Methods, 2 (2010) 1737-1744. [27] M. S. Rizk, H. A. Merey, S. M. Tawakkol, M. N. Sweilam, Anal. Chem. Let., 4 (2014)132-145. [28] H. M. Lotfy, M. A. Hegazy, M. R. Rezk,  Y. R. Omranet,  Acta. Part A: Mol. Bio.Spec., 126 (2014) 197–207. [29] M. M. Ayad, A. Shalaby, H. E. Abdellatef,  M. M. Hosny, Anal. Bioanal. Chem., 375 (2003) 556-560. [30] A. F. Alghamdi, Amer. J. Anal. Chem., 2 (2011) 174-181. [31] M. H. Türkdemir, G. Erdögdu, T. Aydemir, A. A. Karagözler, E. Karagözler, J. Anal. Chem., 56 (2001) 1047-1050.  [32] A. F. Al-Ghamdi, F. Kooli, J. Mater. Environ. Sci., 4 (2013) 762-769. 

https://revues.imist.ma/index.php/morjchem/article/download/5175/3520

HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC DETERMINATION OF GLIMEPIRIDE AND TIMOLOL IN PHARMACEUTICAL FORMULATIONS USING BRITTON -ROBINSON BUFFER IN MOBILE PHASES

Abstract

Similar works

Full text

Available Versions

Revues Scientifiques Marocaines

Revues Scientifiques Marocaines