Saksagliptin ve dapagliflozinin sabit doz kombinasyonu antidiyabetik ilaç tedavisi için onaylanmıştır ve Qtern markası ile pazarda yer almaktadır. Bu çalışmada amaç, insan plazmasındaki saksagliptin ve dapagliflozinin eş zamanlı tayini için Avrupa Gıda ve İlaç İdaresi kılavuzlarına uygun şekilde linagliptin iç standardı kullanarak ve basit, hızlı, hassas ve validasyonu yapılmış izokroatik ters faz-yüksek performanslı sıvı kromatografi (RP-HPLC) yöntemi geliştirmektir. Gereç ve Yöntemler: Method 4'lü akış pompasına sahip Waters 2695 marka HPLC cihazı ile gerçekleştirilmiştir. Analitin ayrılmasında Eclipse XDB C18 kolon (150 × 4.6 mm × 5 μm) kullanılmıştır. Kullanılan mobil fazın bileşimi ise pH 5.0 ayarlanmış %0.1 orto fosforik asit ve asetonitril (50:50) şeklinde olup akış hızı analiz süresince 1 mL/dk'dır. Bulgular: Analit 254 nm'de tayin edilmiştir. İç standart, saksagliptin ve dapagliflozinin alıkonma zamanları sırasıyla 2.746, 5.173 ve 7.218 dk olarak tespit edilmiştir. Pikler interferanslar gözlenmeden elde edilmiştir. Metot validasyonu saksagliptin ve dapagliflozin için sırasıyla 0.01 ile 0.5 μg/mL ve 0.05 ile 2 μg/mL doğrusal derişim aralığında 0.998 korelasyon katsayısı ile gerçekleştirilmiştir. Numunlere ait 6 ölçüme ait kesinlik ve doğruluk tayin sınırları içerisinde bulunmuştur. Analitlerin insan plazması içinde-28°C sıcaklıkta 37 gün boyunca kararlı halde kaldığı belirlenmiştir.
A rapid, sensitive, and accurate high performance liquid chromatography for the determination of axitinib (AN) in rabbit plasma is developed using crizotinibe as an internal standard (IS). Axitinib is a tyrosine kinase inhibitor, used in the treatment of advanced kidney cancer, which works by slowing or stopping the growth of cancer cells. The chromatographic separation was performed on a Waters 2695, Kromosil (150 mm × 4.6 mm, 5 µm) column using a mobile phase containing buffer (pH 4.6) and acetonitrile in the ratio of 65:35 v/v with a flow rate of1 mL/min. The analyte and internal standard were extracted using liquid-liquid extraction with acetonitrile. The elution was detected by photo diode array detector at 320 nm.The total chromatographic runtime is 10.0 min with a retention time for axitinib and IS of 5.685, and 3.606 min, respectively. The method was validated over a dynamic linear range of 0.002-0.2µg/mL for axitinib with a correlation coefficient of r 2 0.999.
Objective: The proposed method aims to develop a simple, rapid, sensitive and validated isocratic reversed-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of linagliptin and empagliflozin in human plasma.Methods: Chromatography was performed on waters 2695 HPLC equipped with a quaternary pump. The separation was carried using discovery C18 (250×4.6×5) column, buffer: acetonitrile (68:32) as mobile phase with 1 ml/min flow rate. The analyte detection was monitored at 218 nm.Results: Retention time of linagliptin, empagliflozin and internal standard was found at 6.421, 4.696, and 4.074 min respectively. The peaks were found to be free of interference. The method is validated over a dynamic linear range of 0.01-10.0 µg/ml for both drugs with a correlation coefficient of 0.998. The precision and accuracy of samples of six replicate measurements at lower limit of quantification (LLOQ) level were within the limit. The analytes were found to be stable in human plasma at-28 °C for 37 d.Conclusion: The stability, sensitivity, specificity and reproducibility of this method make it suitable for the determination of linagliptin and empagliflozin in human plasma.
Objective: The fixed dose combination of saxagliptin and dapagliflozin, recently approved antidiabetic medication. It is marketed with a brand name Qtern. The intend method aim to develop a simple, rapid, sensitive and validated isocratic reversed phase high performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of saxagliptin and dapagliflozin in human plasma by using linagliptin as internal standard as per US-FDA guidelines. Materials and methods: The method was performed on Waters 2695 HPLC equipped with quaternary pump. The analyte separation was achieved using Eclipse XDB C18 (150×4.6µ×5mm) column with a mobile phase consisting of 0.1% ortho phosphoric acid and acetonitrile (50:50) with pH adjusted to 5.0 at1ml/min flow rate. Results: The analyte was detected at 254nm. Retention time of the internal standard, saxagliptin and dapagliflozin was found at 2.746, 5.173 and 7.218minutes, respectively. The peaks were found to be free of interference. The method is validated over a dynamic linear range of 0.01 to 0.5µg/ml and 0.05 to 2µg/ml for saxagliptin and dapagliflozin, respectively, with a correlation coefficient of 0.998. The precision and accuracy of samples of six replicate measurements at LLOQ level was within limit. The analytes were found to be stable in human plasma at-28°C for 37 days. Conclusion: The stability, sensitivity, specificity and reproducibility of this method make it appropriate for the determination of saxagliptin and dapagliflozin in human plasma.
Soil being a major reservoir for microorganisms it is a source of interest for isolation of antibiotic producing organisms. The emergence of antibiotic resistance and need for better, broad spectrum antibiotics is always in high demand. In the present study, antibiotic producing bacteria were isolated from a local soil sample. Total ten soil samples were collected from local pond aseptically and subjected to serial dilution. Crowded plate technique was employed for the isolation of the colony. Total five isolated were isolated which exhibited zone of inhibition around the colony. The isolated colonies were subjected to morphological, microscopical and biochemical characterization. All five colonies were found to be gram positive, non-sporulating organisms and found they belong to the Actinobacteria class. The isolated colonies were subjected to screening for antimicrobial activity against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis and Yeast by perpendicular streak method. The primary screening results conclude that except one colony all have good antimicrobial activity. One colony found to be highly potential activity which had inhibition towards gram positive, gram negative, sporulating and fungal activity. This study may contribute in providing information on the antibiotic producing microorganisms in soil. Further characterization, purification, and structural elucidation are recommended to know the novelty, quality and commercial value of these antibiotics.
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