Establishment and Application of HPLC Method for Content Determination of Rapamycin in Human Monocyte THP-1 Derived Foam Cells / 中国药房

ABSTRACT

OBJECTIVE:To establish the method for the content determination of rapamycin (RAPA) in human monocyte THP-1 derived foam cells,and to study the effects of RAPA targeting preparation(RAPA-NP-Apt)targeting at foam cells. METH-ODSFoam cells model were established through THP-1 cells were induced by oxidized low density lipoprotein. Foam cells were incubated with 200 ng/mL RAPA or 200,400,800 ng/mL RAPA-NP-Apt for 60 min. The content of RAPA was determined by HPLC. The determination was performed on Diamonsil C18 column with mobile phase consisted of acetonitrile-water(9010,V/V) at flow rate of 1.0 mL/min. The column temperature was set at 40 ℃,and the detection wavelength was 278 nm. The sample size was 20 μL. RESULTS:The concentration of RAPA ranged 50-6400 ng/mL (r=0.99996) with average recovery of 98.72%(RSD=0.62%,n=3). RSDs of inter-day and intra-day were not more than 6.15%(n=6),RSD of stability was lower than 2%(n=6),and RSD of repeatability was 1.64%(n=6). After foam cells were incubated with RAPA or low-concentration,medi-um-concentration and high-concentration of RAPA-NP-Apt,the contents of RAPA were 12,43,98,140 ng/106 cells. CONCLU-SIONSThe method is simple,stable and reproducible. It can be used for content determination of RAPA in foam cells. RA-PA-NP-Apt can improve the effects of RAPA targeting at foam cells.