Assar N.H.Hamouda H.M.Mohamed G.S.Amin H.M.Department of MicrobiologyNational Organization for Drug Control and ResearchEgypt; Faculty of PharmacyCairo UniversityEgypt; Department of MicrobiologyM. S. A UniversityEgypt; Department of Microbiology and ImmunologyKing Abdul-Aziz UniversitySaudi Arabia2020-01-092020-01-092015[1] Beebe, F.A; Barkin, R.L; Barkin, S.A. Clinical and pharmacologic review of skeletal muscle relaxants for musculoskeletal conditions. Am. J. Ther., 2005, 12(2), 151-171. [2] British Pharmacopoeia; Her Majesty Stationery Office, London, 2011. [3] Martindale. The Complete Drug Reference, 35th ed.; Pharmaceutical Press, London, 2007. [4] Hollifield, R.D; Conklin, J.D. Determination of dantrolene in biological specimens containing drug-related metabolites. J. Pharma. Sci., 1973, 62(2), 271-274. [5] Saxena, S.J; Honigberg, I.L; Stewart, J.T; Keene, G.R; Vallner, J.J. Liquid chromatography in pharmaceutical analysis VI: Determination of dantrolene sodium in a dosage form. J. Pharma. Sci., 1977, 66(2), 286-288. [6] Saxena, S.J; Honigberg, I.L; Stewart, J.T; Vallner, J.J. Determination of dantrolene sodium in biological fluids. J. Pharma. Sci., 1977, 66(5), 751-753. [7] Hackett, L.P; Dusci, L.J. Determination of dantrolene sodium in human plasma using high-performance liquid chromatography. J. Chromatogr. A, 1979, 179(1), 222-224. [8] Katogi, Y.; Tamaki, N.; dachi, M.A; Terao, J.; Mitomi, M. Simultaneous determination of dantrolene and its metabolite, 5- hydroxydantrolene in human plasma by high-performance liquid chromatography. J. Chromatogr. B, 1982, 231, 404-408. [9] Wuis, E.W; Grutters, A.C; Vree, T.B; Kleyn, E.V.D. Simultaneous determination of dantrolene and its metabolites, 5- hydroxydantrolene and nitro-reduced acetylated dantrolene (F 490), in plasma and urine of man and dog by high-performance liquid chromatography. J. Chromatogr. B, 1982, 231(2), 401-409. [10] Lalande, M.; Mills, P.; Peterson, R.G. Determination of dantrolene and its reduced and oxidized metabolites in plasma by highperformance liquid chromatography. J. Chromatogr. B, 1988, 430(1), 187-191. [11] Wuis, E.W; Janssen, M.G; Vree, T.B; van der Kleijn, E. Determination of a dantrolene metabolite, 5-(p-nitrophenyl)-2-furoic acid, in plasma and urine by high-performance liquid chromatography. J. Chromatogr. B, 1990, 526(2), 575-580. [12] Cox, P.L; Heotis, J.P; Polin, D.; Rose, G.M. Quantitative determination of dantrolene sodium and its metabolites by differential pulse polarography. J. Pharma. Sci., 1969, 58(8), 987-989. [13] Ghoneim, E.M. Electroreduction of the muscle relaxant drug dantrolene sodium at the mercury electrode and its determination in bulk form and pharmaceutical formulation. Chem. Pharm. Bull., 2007, 55(10), 1483-1488. [14] Khan, S.R; Tawakkul, M.; Sayeed, V.A; Faustino, P.; Khan, M.A. Stability characterization, kinetics and mechanism of degradation of dantrolene in aqueous solution. Sci. Res., 2012, 3(3), 281-290. [15] Bouhsain, Z.; Garrigues, S.; Morales-Rubio, A.; Guardia, M. Flow injection spectrophotometric determination of paracetamol in pharmaceuticals by means of on-line microwave-assisted hydrolysis and reaction with 8-hydroxyquinoline (8-quinolinol). Anal. Chim. Acta, 1996, 330(1), 59-69. Qualitative and Quantitative Chemometry as Stability-Indicating Methods Current Pharmaceutical Analysis, 2018, Vol. 14, No. 1 67 [16] Rodenas, V.; Garcia, M.S; Sanchez-Pedreno, C.; Albero, M.I. Simultaneous determination of propacetamol and paracetamol by derivative spectrophotometry. Talanta, 2000, 52(3), 517-523. [17] Bloomfield, M.S. A sensitive and rapid assay for 4-aminophenol in paracetamol drug and tablet formulation by flow injection analysis with spectrophotometric detection. Talanta, 2002, 58(6), 1301- 1310. [18] Filik, H.; Hayvali, M.; Kilic, E. Sequential spectrophotometric determination of paracetamol and p-aminophenol with 2, 2-(1, 4- phenylenedivinylene) bis-8-hydroxyquinoline as a novel coupling reagent after microwave assisted hydrolysis. Anal. Chim. Acta, 2005, 535(1-2), 177-182. [19] Abdellatef, H.E; Ayad, M.M; Soliman, S.M; Youssef, N.F. Spectrophotometric and spectrodensitometric determination of paracetamol and drotaverineHCl in combination. Spectrochim. Acta A, 2007, 66(4-5), 1147-1151. [20] El-Yazbi, F.A; Hammu, H.H; Assi, S.A. Derivative-ratio spectrophotometric method for the determination of ternary mixture of aspirin, paracetamol and salicylic acid. Spectrochim. Acta A, 2007, 68(2), 275-278. [21] Khoshayand, M.R; Abdollahi, H.; Shariatpanahi, M.; Saadatfard, A.; Mohammadi, A. Simultaneous spectrophotometric determination of paracetamol, ibuprofen and caffeine in pharmaceuticals by chemometric methods. Spectrochim. Acta A, 2008, 70(3), 491-499. [22] Yehia, A.M; Mohamed, K. Abd El-Rahman. Application of normalized spectra in resolving a challenging Orphenadrine and Paracetamol binary mixture. Spectrochim. Acta A, 2015, 138, 21- 30. [23] Celma, C.; Allué, J.A.; Pruñonosa, J.; Peraire, C.; Obach, R. Simultaneous determination of paracetamol and chlorpheniramine in human plasma by liquid chromatography-tandem mass spectrometry. J. Chromatogr. A, 2000, 870(1-2), 77-86. [24] Vignaduzzo, S.E; Kaufman, TS. Development and validation of a HPLC method for the simultaneous determination of bromhexine, chlorphenramine, paracetamol and pseudoephedrine in their combined cold medicine formulations. J. Liq. Chromatogr. R. T., 2013, 36(20), 2829-2843. [25] Lourenção, B.C; Medeiros, R.A; Rocha-Filho, R.C; Mazo L.H; Fatibello-Filho, O. Simultaneous voltammetric determination of paracetamol and caffeine in pharmaceutical formulations using a boron-doped diamond electrode. Talanta., 2009, 78(3), 748-752. [26] Atta, N.F, Galal, A.; Azab, S.M. Electrochemical determination of paracetamol using gold nanoparticles – application in tablets and human fluids. Int. J. Electrochem. Sci., 2011, 6, 5082-5096. [27] Rashed, N.S; Abdallah, O.M; Farag, R.S; Awad, S.S. Validated bivariate calibration spectrophotometric and high performance liquid chromatographic methods for simultaneous determination of dantrolene sodium and paracetamol in pharmaceutical dosage form. Adv. Anal. Chem., 2014, 4(1), 1-8. [28] El-Bagary, R.I; Elkady, E.F; Hegazi, M.A; Amin, N.E. Spectrophotometric methods for the simultaneous determination of paracetamol and dantrolene sodium in pharmaceutical dosage form. Eur. J. Chem., 2014, 5(1), 96-100. [29] Salem, H.; Mohamed, D. A comparative study of smart spectrophotometric methods for simultaneous determination of a skeletal muscle relaxant and an analgesic in combined dosage form. Spectrochim. Acta A, 2015, 140, 166-173. [30] Hadad, G.M; Emara, S.; Mahmoud, W.M. Development and validation of a stability-indicating RP-HPLC method for the determination of paracetamol with dantrolene or/and cetirizine and pseudoephedrine in two pharmaceutical dosage forms. Talanta, 2009, 79(5), 1360-1367. [31] El-Saharty, Y.S; Riad, S.M; Yehia, A.M; Sami, I. Stabilityindicating chromatographic methods for the determination of a skeletal muscle relaxant and an analgesic in their combined dosage form. J. Chromatogr. Sep. Tech., 2016, 7, 2. [32] Shehata, M.A; Ashour, A.; Hassan, N.Y; Fayed, A.S; El-Zeany, B.A. Liquid chromatography and chemometric methods for determination of rofecoxib in presence of its photo degradation and alkaline degradation products. Anal. Chim. Acta, 2004, 519(1), 23- 30. [33] Hoang, V.D.; Nhung, N.P.; Aboul-Enein, H.Y. Recent developments and applications of derivative spectrophotometry in pharmaceutical analysis. Curr. Pharm. Anal., 2013, 9(3), 261-277. [34] Hegazy, M.A.; Yehia, A.M.; Moustafa, A.A. Bivariate versus multivariate smart spectrophotometric calibration methods for the simultaneous determination of a quaternary mixture of mosapride, pantoprazole and their degradation products. Pharmazie, 2013, 68, 317-326. [35] Yehia, A.M.; Arafa, R.M.; Abbas, S.S.; Amer, S.M. Ratio manipulating spectrophotometry versus chemometry as stability indicating methods for cefquinome sulfate determination. Spectrochim. Acta A, 2016, 153, 231-240. [36] Yehia A.M.; Mohamed H.M.. Chemometrics resolution and quantification power evaluation: Application on pharmaceutical quaternary mixture of Paracetamol, Guaifenesin, Phenylephrine and p aminophenol. Spectrochim. Acta A, 2016, 152, 491-500. [37] El-Ragehy, N.; Yehia, A.M; Hassan, N.; Tantawy, M.; Abdelkawy, M. Chemometrics tools in detection and quantitation of the main impurities present in aspirin/dipyridamole extended-release capsules. J. AOAC Int., 2016, 99, 948-956. [38] Brereton, R.G. Multilevel multifactor designs for multivariate calibration. Analyst., 1997, 122(12), 1521-1529. [39] Rajalahti, T.; Kvalheim, O.M. Multivariate data analysis in pharmaceutics: A tutorial review. Int. J. Pharm., 2011, 417(1-2), 280- 290. [40] De Juan, A.; Tauler, R. Chemometrics applied to unravel multicomponent processes and mixtures: Revisiting latest trends in multivariate resolution. Anal. Chim. Acta, 2003, 500(1-2), 195-210. [41] Windig, W.; Guilment, J. Interactive self-modeling mixture analysis. Anal. Chem., 1991, 63(14), 1425-1432. [42] International Conference on Harmonization, Validation of Analytical Procedures: Methodology (Q2B); 1996.9755071https://doi.org/https://t.ly/52wGPScopusMSA Google Scholar192 Staphylococcus samples were isolated from patients in Egyptian hospitals and 112 were identified as Staphylococcus aureus using conventional methods based on morphological and biochemical characteristics. Sensitivity of isolates to a range of antibiotics was also tested by disc diffusion method, which revealed that 82% of isolates were MRSA isolates. Methicillin resistance was confirmed using PCR-based molecular approach, 8 isolates were harboured mecA gene and 4 isolates were Borderline S. aureus (BORSA). MICs of different antibiotic classes were determined; results were varied to different antibiotics, where Penicillin and Ampicillin MICs varied from resistant to borderline resistant isolates (512,62?g/ml). Cinnamomum cassia, Syzygium aromaticum, Glycyrrhiza glabra, Rosmarinus officinalis and Salvia officinalis plant extracts were tested to examine their antibacterial activity against MRSA isolates. MIC of tested plant extracts was evaluated by agar dilution method. Diethyl ether extracts of G. glabra, R. officinalis and S. officinalis showed the lowest MICs values (0.05, 0.39, 0.195mg/ml respectively). Investigation of possible synergistic effect upon combination between plant extracts with the lowest MICs values was evaluated by a checkerboard titration assay. Combination between diethyl ether extracts of G. glabra and R. officinalis at conc. 0.0125 and 0.0975 mg/ml respectively showed synergistic effect on MRSA isolates and standard strain. Both extracts shown antibacterial effects on MRSA cells by shrinkage of the protoplasts and disruption of the cytoplasmic membrane evidenced by protein analysis and Microscopic examination of cells pre and post treatment by TEM .These results suggest that these extracts might be used as a promising antibacterial agent.EnglishOctober University for Modern Sciences and Artsجامعة أكتوبر للعلوم الحديثة والآدابUniversity of Modern Sciences and ArtsMSA UniversityAgar well diffusion methodGlycyrrhiza glabraMinimum inhibitory concentrationRosmarinus officinalisSynergistic effectamikacinampicillinantiinfective agentazithromycincefalexincefepimecefotaximecefuroximechloramphenicolCinnamomum cassia extractciprofloxacinclove extractdoxycyclineerythromycingentamicinGlycyrrhiza glabra extractimipenemlevofloxacinmeticillinnalidixic acidneomycinnorfloxacinoxacillinpenicillin derivativeplant extractRosmarinus officinalis extractSalvia officinalis extractsulfonamidetrimethoprimunclassified drugunindexed drugagar dilutionantibacterial activityantibiotic resistanceantibiotic sensitivityArticlebacterial genebacterial membranebacterial strainbacterium isolateclovecontrolled studydisk diffusiondrug potencydrug potentiationEgyptmecA genemembrane damagemethicillin resistant Staphylococcus aureusminimum inhibitory concentrationmultidrug resistancenonhumanprotoplastArticlehttps://doi.org/