Bempong Daniel K, Bhattacharyya Lokesh
Information and Standards Development, US Pharmacopeia, 12601 Twinbrook Parkway, Rockville, MD 20852, USA.
J Chromatogr A. 2005 May 6;1073(1-2):341-6. doi: 10.1016/j.chroma.2004.12.061.
An isocratic RP-HPLC procedure has been developed and validated for the quantitative determination of ketoprofen in a topical gel. The HPLC procedure consist of a YMC ODS-AQ, 5-microm particle size analytical column (150 mm x 4.6 mm); Alltech Econosphere C18, 5-microm particle size guard column; detection at 233 nm; 1 ml/min flow rate; 20-microl injection volume. The mobile phase consisted of pH 3.5 phosphate buffer-water-acetonitrile (8:43:49, v/v). Sample preparation was a simple extraction of ketoprofen with mobile phase. The above conditions resolved and eluted ketoprofen, excipients, and potential degradants within 35 min, with ketoprofen eluting at about 6.5 min. The procedure was validated with respect to specificity, accuracy, precision, and linearity. The accuracy of the procedure, determined by spike recovery measurements, was 100.1-100.5%. The intra- and inter-day precisions were demonstrated by the relative standard deviations (RSD) of 0.3-0.6% and 0.5%, respectively. The intermediate precision was determined by comparing the results obtained with four independently prepared samples by two chemists using two columns on different days. The results indicate no significant difference (P = 0.17). The procedure showed linearity over the concentration range 4 x 10(-5) to 1 x 10(-1) mg/ml.
已开发并验证了一种等度反相高效液相色谱法（RP-HPLC），用于定量测定局部用凝胶中的酮洛芬。该高效液相色谱法包括一根YMC ODS-AQ、粒径为5微米的分析柱（150毫米×4.6毫米）；Alltech Econosphere C18、粒径为5微米的保护柱；检测波长为233纳米；流速为1毫升/分钟；进样体积为20微升。流动相由pH值为3.5的磷酸盐缓冲液-水-乙腈（8:43:49，v/v）组成。样品制备是用流动相简单萃取酮洛芬。上述条件在35分钟内分离并洗脱了酮洛芬、辅料和潜在降解产物，酮洛芬在约6.5分钟时洗脱。该方法在特异性、准确性、精密度和线性方面进行了验证。通过加样回收率测量确定的该方法的准确性为100.1-100.5%。日内和日间精密度分别通过相对标准偏差（RSD）为0.3-0.6%和0.5%来证明。中间精密度是通过比较两位化学家在不同日期使用两根柱子独立制备的四个样品所获得的结果来确定的。结果表明无显著差异（P = 0.17）。该方法在4×10^(-5)至1×(10)^(-1)毫克/毫升的浓度范围内呈线性。
