Université catholique de Louvain, Louvain Drug Research Institute, Unité de Pharmacie Galénique, UCL 73.20, Avenue Emmanuel Mounier 73, 1200 Brussels, Belgium.
J Pharm Biomed Anal. 2010 Feb 5;51(3):633-9. doi: 10.1016/j.jpba.2009.09.040. Epub 2009 Oct 2.
An HPLC/UV method was first optimized for the separation and quantitation of human acylated and unacylated (or des-acyl) ghrelin from aqueous solutions. This method was validated by an original approach using accuracy profiles based on tolerance intervals for the total error measurement. The concentration range that achieved adequate accuracy extended from 1.85 to 59.30microM and 1.93 to 61.60microM for acylated and unacylated ghrelin, respectively. Then, optimal temperature, pH and buffer for sample storage were determined. Unacylated ghrelin was found to be stable in all conditions tested. At 37 degrees C acylated ghrelin was stable at pH 4 but unstable at pH 7.4, the main degradation product was unacylated ghrelin. Finally, this validated HPLC/UV method was used to evaluate the binding of acylated and unacylated ghrelin to liposomes.
首次优化了 HPLC/UV 方法，以分离和定量来自水溶液的酰化和非酰化（或去酰化）人 ghrelin。该方法通过使用基于总误差测量公差区间的准确度概况的原始方法进行了验证。获得足够准确度的浓度范围分别为 1.85 至 59.30μM 和 1.93 至 61.60μM，用于酰化和非酰化 ghrelin。然后，确定了用于样品储存的最佳温度、pH 值和缓冲液。在所有测试条件下，非酰化 ghrelin 均稳定。在 37°C 时，酰化 ghrelin 在 pH 4 下稳定，但在 pH 7.4 下不稳定，主要降解产物为非酰化 ghrelin。最后，使用验证后的 HPLC/UV 方法评估了酰化和非酰化 ghrelin 与脂质体的结合。
