title: HPTLC method for direct determination of gemifloxacin mesylate in human plasma.
abstract: Novel, simple and sensitive high performance thin-layer chromatography (HPTLC) with fluorescence detection has been successfully developed and validated for determination of gemifloxacin mesylate (GFX) in plasma samples without prior pretreatment. Montelukast (MK) was used as internal standard. GFX and MK in plasma samples were separated using a mobile phase consisting of a mixture of ethyl acetate:methanol:25% ammonia, (8:4.5:3, v/v/v). The emission intensity was measured using optical filter K400 after excitation at 342 nm. The Rf values for GFX and MK were 0.45±0.03 and 0.79±0.02, respectively. Under the optimum conditions, a linear relationship with good correlation coefficient (r=0.9965, n=6) was obtained in concentration range of 3-180 ng/band. The LOD and LOQ of the proposed method were 0.45 and 1.5 ng/band, respectively. The accuracy of the method was proved as the recovery % of GFX from spiked human plasma was 94.21-101.85%. The efficiency of the proposed method was confirmed by in-vivo application on human plasma in real patient samples. Moreover, the stability of GFX in plasma was carefully tested at different conditions and compared to others in aqueous solution.
authors.affiliation: Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Assiut University, Assiut, Egypt. Electronic address: wesamelkosi@yahoo.com.
authors.forename: W M
authors.initials: WM
authors.lastname: El-Koussi
authors.affiliation: Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Assiut University, Assiut, Egypt.
authors.forename: N N
authors.initials: NN
authors.lastname: Atia
authors.affiliation: Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Assiut University, Assiut, Egypt; Department of Pharmaceutical Chemistry, College of Pharmacy, Najran University, Najran, Saudi Arabia.
authors.forename: A M
authors.initials: AM
authors.lastname: Mahmoud
authors.affiliation: Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Assiut University, Assiut, Egypt.
authors.forename: S R
authors.initials: SR
authors.lastname: El-Shabouri
mesh_terms: D002855:Chromatography, Thin Layer / Q000379:methods*; D004355:Drug Stability; D024841:Fluoroquinolones / Q000097:blood* / Q000737:chemistry; D000077735:Gemifloxacin; D006801:Humans; D016014:Linear Models; D009287:Naphthyridines / Q000097:blood* / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
chemical_list: D024841:Fluoroquinolones; D009287:Naphthyridines; D000077735:Gemifloxacin
keywords: Fluorescence detection; Gemifloxacin mesylate; HPTLC; Human plasma