叶酸受体介导pH敏感型分子靶向阿霉素脂质体的表征与含量测定法的建立

Characterization and content determination of folate receptor-mediated pH-sensitive molecule-targeted doxorubicin liposome containing F-PEOz-DSPE

目的:表征叶酸受体介导pH敏感型分子靶向阿霉素脂质体,建立RP-HPLC法进行含量测定。方法:采用动态光散射法测定脂质体的粒径;采用液相色谱法开展含量测定的方法学研究与验证。选用Agilent HC-C18色谱柱(250 mm×4.6 mm,5μm),流动相为1.23%醋酸钠水溶液(用冰醋酸调pH至3.3)-甲醇(55∶45),等度洗脱,流速为1.0 mL.min-1,检测波长233 nm,柱温30℃。结果:自制的新型脂质体粒径(120±20)nm(PDI<0.200),包封率不小于91.0%。建立的RP-HPLC法中,自制脂质体的辅料对主峰无干扰,且各杂质峰与主成分峰均能达到基线分离,检测限为0.2 ng(S/N=3),定量限为1.0 ng(S/N=10),主成分的浓度在1.0～40.0μg.mL-1范围内线性良好(r=0.9991)。结论:本文所建方法简便易行,专属性强,灵敏,耐用性好,适于本脂质体中阿霉素的含量测定。

Objective：To characterize the folate receptor- mediated pH -sensitive molecule -targeted doxorubicin liposome containing F - PEOz - DSPE and establish an HPLC assay method of the detection for the novel lipo- some. Methods： The Agilent HC - Cls chromatographic column （ 250 mm × 4.6 mm, 5 p,m） were used. The mobile phase consisted of a mixture of 1.23% sodium acetate aqueous solution（ pH adjusted to 3.3 using acetic acid） - methanol（55：45） with a flow rate of 1.0 mL · rain-1 by isocratic elution;the UV detection was carried out at a wavelength of 233 nm. The column oven temperature was maintained at 30 ℃. Results： The average particle size of the novel liposome was （ 120 ± 20 ） nm （ PDI 〈 0. 200）, the encapsulation efficiency was more than 91.0%. LOD （ limit of detection） of the method was 0. 2 ng （ S/N= 3 ）, LOQ （ limit of quantitation） was 1.0 ng （ S/N= 10 ）. The established RP - HPLC showed that the excipient of the self - made liposome had no interference with the main peak, and each impurity peak was well baseline separated with the main component peak. The calibration curve of the main component concentration was linear within the range of 1.0 -40.0 μg · mL- 1 （r = 0. 9991 ）. Conclusion： This method is simple and practical with strong specificity and good durability. It is suitable for detection of doxoru- bicin in this liposome containing F- PEOz -DSPE.