Waters HPLC alliance e2695 system with PDA detector with a reverse phase C18 column (Symmetry®, 5 μm, 4.6 × 250 mm; Waters Technology Corporation, USA) was used for analysis of FLB at wavelength of 247 nm. The mobile phase was acetonitrile and water (pH adjusted to 6 using acetic acid), at a ratio of 70:30 (v/v), flow rate of 0.5 ml/min, retention time of 2.5 min, and injection volume of 50 μl. Data acquisition and analysis were performed using Empower software (Waters Corporation, MA). The calibration curve (peak area vs. concentration) was generated over a range of 5–80 μg/ml and was found to be linear with a correlation coefficient R2 = 0.9974. LOD and LOQ was found to be 1.31 μg/ml and 3.97 μg/ml, respectively [33 (link), 34 (link)].
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HPLC Analysis of FLB in Pharmaceuticals
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Other organizations : Florida Agricultural and Mechanical University
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Variable analysis
independent variables
- Ratio of acetonitrile and water (pH adjusted to 6 using acetic acid) in the mobile phase
- Flow rate of the mobile phase
- Injection volume
- Peak area
- Concentration of FLB
- HPLC system with PDA detector
- Reverse phase C18 column (Symmetry®, 5 μm, 4.6 × 250 mm; Waters Technology Corporation, USA)
- Wavelength of 247 nm for analysis
- Retention time of 2.5 min
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