摘要
建立了一种分离、测定磷脂酰胆碱(PC)、磷脂酸(PA)、心磷脂(CL)和磷脂酰甘油(PG)的等度RPHPLC法。色谱条件为:在PEC18柱上,用V(甲醇)∶V(乙腈)∶V(水)=79∶8∶13的溶液作流动相,以2.0mLmin的流速进行等度洗脱,紫外检测波长205nm,谱带宽5nm。以熊胆为样品,经预处理后用所建立的方法进行分析,结果表明:6个熊胆样品中PC的质量比差别较大;PA的质量比除2号样品较低外,其余相近;PG除2号样品外都能检出,其中6号样品中PG的质量比稍大。PC的平均回收率为89.21%,RSD为2.1%,重现性较好,分析时间短,在12min内即可将上述4种组分分离。该方法对进一步分离、测定磷脂类化合物具有一定的参考价值。
Linear calibration curves were obtained for 4.5 10.5 μg of phosphatidylcholine(PC), 1.5 3.5 μg of phosphatidic acid(PA), 9.0 21.0 μg of cardiolipin(CL), 2.25 5.25 μg of phosphatidylglycerol(PG) by reversed phase high performance liquid chromatography with UV detector at 205 nm, P E C 18 column (15 cm×0.46 cm i.d.,5 μm) and isocratic elution with mobile phase of methanol acetonitrile water (79∶8∶13,V/V/V) at a flow rate of 2.0 mL/min. Separation and detection of these four standards were achieved reproducibly and easily in less than 12 minutes. The RSDs of peak area measurements were from 0.77% to 8.86 %. Average recovery of PC was 89.21%(RSD=2.1%). The air dried bear biles were ground into powder and extracted continuously with chloroform methanol (1∶1,V/V) to a final dilution 25 folds the volume of the sample. The extract was washed by addition of half its volume of 0.58% aqueous NaCl. The resulting mixture separated into two phases. The lower phase was the total pure lipid extract and was dried at room temperature under nitrogen. The lipid residue was dissolved in methanol to the desired volume and injected into the HPLC system for analysis. The results indicated that six kinds of bear biles had differences in the phospholipid composition. The method enables a simple, rapid and reproducible quantification of PC, PA, PG and CL.
出处
《色谱》
CAS
CSCD
北大核心
2000年第1期57-60,共4页
Chinese Journal of Chromatography
关键词
熊胆
磷脂类化合物
HPLC
reversed phase high performance liquid chromatography
phosphatidylcholine
phosphatidic acid
cardiolipin
phosphatidylglycerol
bear bile
