Objectives: This study is initiated to develop a simple and reproducible liquid chromatographic method with ultraviolet detection for the estimation of Sisomicin in Gentamicin pharmaceutical formulation by application of total error statistical tool. Methods: Gentamicin has a weak UV chromophore it is not possible to detect low levels of known related substance of gentamicin using a UV detector. Hence derivatization technique was applied to detect those substances. Chromatographic separation was accomplished by using Thermo scientific Hypersil Gold column (150 x 4.6 mm) and 5µm particle size as stationary phase by isocratic elution with Methanol: Water: Glacial acetic acid: Sodium hexane sulfonate in the ratio 70:25:5:3% v/v/ v/w as mobile phase. Sisomicin was detected at 330 nm within 25 min with a flow rate of 0.5 mL/min. Results: Concurrent results were obtained in the developed analytical method based on total error measurement. Proposed method showed good linearity response (r> 0.995) with limit of quantification at 0.007495 mg/mL, % relative standard deviation less than 1% in repeatability and % recovery was found within 96 to 98 % in accuracy. Accuracy profile result found within the range of ±10% and risk profile ±5% between the two set. Conclusion: This method can separate all the analogues of Gentamicin including known related substances. The finding demonstrated that method could be suitable for quantification of related substances in parental liquid dosage form.
Aminoglycosides antibiotics are considered to be the antimicrobial agents used frequently in the treatment of human diseases caused by a bacterial infection. Most of the aminoglycosides antibiotics are highly polar in nature and they are lacking the UV absorbing chromophore in the molecules. The present articles accentuate the analytical method associated with the analysis of aminoglycosides molecules. Various chromatographic techniques like liquid chromatography, gas chromatography; mass spectrometry were used for the detection of aminoglycosides antibiotics. However, due to its limitation in the ultraviolet-visible spectrophotometry (UV/Vis) technique, different types of detection techniques like corona-charged aerosol detector (CAD), electrochemical detector (ECD) were used as a most powerful and versatile technique for the demonstration of these molecules in the analytical field. Analytical methods help to ensure the quality of the drug products. This review paper is devoted to providing an overview of the key performance technique used for the application and detection of these aminoglycosides molecules.
A simple reverse-phase high performance liquid chromatographic method (RP-HPLC) was developed for quantification of the related compounds in Cyclophosphamide. The chromatographic separation was achieved with C18 column (250 × 4.6 mm, 5 µm particle size) with gradient elution composed of 0.02 M Potassium dihydrogen phosphate (pH-7.0) as mobile phase-A and 60:40% v/v Acetonitrile / Water as mobile phase-B at a flow rate of 0.8 mL /min and column compartment maintained at 40°C for separation. Detection was carried out at 195 nm. The correlation coefficient (≥0.99) shows the linearity response against concentration over the range of Limit of Quantification (LOQ). Precision studies showed the Relative Standard Deviation (RSD) values less than 5% for Cyclophosphamide and its related compounds. The method was substantiated with respect to specificity, precision, linearity, accuracy, limit of quantification, and robustness. The proposed method could be used for routine analysis of Cyclophosphamide formulations.
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