A high-performance liquid chromatography (HPLC) method with ultraviolet (UV) detection was developed and validated for the quantification of lorazepam in oral solution. The chromatographic conditions include an isocratic separation (25% water / 75% methanol at a flow rate of 0.500 ml/min) on a Waters X=Bridge C18 column (150 x 4.6 mm; 3.5-micron particle size). The method was validated using guidance from the United States Pharmacopeia (USP) chapter, including investigation of system suitability, precision, accuracy, linearity, and specificity. The calibration curves on three non-consecutive days met the linearity criteria R2 >0.99. Each chromatogram for 200 mcg/mL calibration samples, designated as 100% assay level, met system suitability criteria of resolution ≥2.0, tailing factor ≤2.0, column efficiency (theoretical plates) ≥2000, and precision of prior metrics % RSD ≤1.0. Three replicates of each concentration, 150 mcg/mL at 75% assay, 200 mcg/mL at 100% assay, and 250 mcg/mL at 125% assay were assessed for precision and accuracy over 3 days. Precision and accuracy were evaluated and met the inter-day (repeatability) criteria % RSD and % Error ≤ 2% and intra-day (intermediate) criteria % RSD and % Error ≤ 5% at the 75%, 100%, and 125% assay levels. To assess for specificity, 200 mcg/mL samples were assessed for degradation after being subjected to heat (>60°C), oxidation (3% H2O2), acidic (0.1M HCl), and basic (0.1M NaOH) environments. Samples from each condition were evaluated for lorazepam recovery at 0, 24, and 48 hours. Most drug loss was observed with the samples subjected to acidic and oxidative environments, with 14.71% and 13.16% drug loss, respectively after 48 hours. This method was developed to support the 30-day stability investigation of lorazepam oral solution when stored in oral syringes at room and refrigerated temperatures.
| Identifer | oai:union.ndltd.org:ETSU/oai:dc.etsu.edu:asrf-2185 |
| Date | 25 April 2023 |
| Creators | Tubolino, Michelle, Sergent, Sophia, Brown, Stacy, Coffey, Tim |
| Publisher | Digital Commons @ East Tennessee State University |
| Source Sets | East Tennessee State University |
| Detected Language | English |
| Type | text |
| Source | Appalachian Student Research Forum |
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