Mercolini Laura, Musenga Alessandro, Saladini Bruno, Bigucci Federica, Luppi Barbara, Zecchi Vittorio, Raggi Maria Augusta
Department of Pharmaceutical Sciences, Faculty of Pharmacy, Alma Mater Studiorum - University of Bologna, Via Belmeloro 6, 40126 Bologna, Italy.
J Pharm Biomed Anal. 2008 Dec 15;48(5):1303-9. doi: 10.1016/j.jpba.2008.09.009. Epub 2008 Sep 11.
A fast and simple method based on LC with fluorescence detection has been developed for the determination of insulin in innovative formulations consisting of microparticles and inserts for oral and nasal drug administration, respectively. A reverse-phase C8 column and a mobile phase composed of pH 3.7, 40 mM sodium sulphate solution and acetonitrile (24%, v/v) were employed. Using isocratic elution at 1.0 mL/min flow, analysis is completed within 7 min. Three different kinds of spray-dried microparticles were analysed, consisting of an insulin loaded core composed of chitosan salts (chitosan succinate, chitosan adipate or chitosan suberate) coated with stearic acid. Nasal inserts consisted of chitosan/hyaluronate polyelectrolyte complexes which were loaded with insulin and freeze-dried. Insulin was extracted from both the oral and nasal formulations using pH 7.4 phosphate buffer. The employment of fluorescence detection (lambda(exc) = 276 nm, lambda(em) = 306 nm) granted high selectivity, with no interference from the matrix. Full method validation was performed with good results in terms of linearity (insulin concentration range 0.10-30.0 microg/mL), LOD (0.03 microg/mL) and LOQ (0.10 microg/mL), precision (R.S.D.%<3.6) and accuracy (recovery percentage>90.0%). Insulin content in innovative formulations, expressed as percentage w/w, resulted to be between 0.90 and 0.97 for oral innovative formulations, while an average value of 342 microg of insulin was found in a single nasal insert, in good agreement with preparative protocols.
已开发出一种基于液相色谱-荧光检测的快速简便方法，用于测定分别由用于口服和鼻腔给药的微粒和插入剂组成的创新制剂中的胰岛素。采用反相C8柱和由pH 3.7、40 mM硫酸钠溶液和乙腈（24%，v/v）组成的流动相。以1.0 mL/min的流速进行等度洗脱，7分钟内即可完成分析。分析了三种不同的喷雾干燥微粒，其由负载胰岛素的核心组成，该核心由壳聚糖盐（壳聚糖琥珀酸盐、壳聚糖己二酸盐或壳聚糖辛二酸盐）组成，并涂有硬脂酸。鼻腔插入剂由负载胰岛素并冻干的壳聚糖/透明质酸盐聚电解质复合物组成。使用pH 7.4的磷酸盐缓冲液从口服和鼻腔制剂中提取胰岛素。采用荧光检测（激发波长λ(exc)=276 nm，发射波长λ(em)=306 nm）具有高选择性，不受基质干扰。在线性（胰岛素浓度范围0.10 - 30.0 μg/mL）、检测限（0.03 μg/mL）和定量限（0.10 μg/mL）、精密度（相对标准偏差R.S.D.%<3.6）和准确度（回收率>90.0%）方面进行了全面的方法验证，结果良好。创新制剂中的胰岛素含量，以w/w百分比表示，口服创新制剂为0.90至0.97，而在单个鼻腔插入剂中发现胰岛素的平均值为342 μg，与制备方案高度一致。
