摘要
目的建立人血浆样品中沙库巴曲、缬沙坦及其主要代谢产物LBQ-657的高效液相色谱-荧光检测(HPLC-FD)分析方法。方法采用乙腈沉淀法处理血浆样品,SN-38作为内标进行内标法定量。采用Agilent Eclipse SB-C_(18)色谱柱(250 mm×4.6 mm,5μm),荧光检测器;以甲醇-磷酸二氢钠缓冲液(pH 3.0)(67∶33)为流动相;流速1.0 mL·min^(-1);柱温25℃;进样量20μL。采用多波长法,在0~6 min,λ_(EX)=380 nm,λ_(EM)=430 nm,定量检测SN-38;6~9.4 min,λ_(EX)=255 nm,λ_(EM)=314 nm,定量检测LBQ-657;9~12 min,λ_(EX)=255 nm,λ_(EM)=374 nm,定量检测缬沙坦;12~18 min,λ_(EX)=255 nm,λ_(EM)=314 nm,定量检测沙库巴曲。5例患者口服100 mg沙库巴曲缬沙坦钠片后1.5,3 h分别采样,检测血浆中沙库巴曲、缬沙坦及LBQ-657浓度。结果人血浆样品中各成分在0.1~20μg·mL^(−1)内线性关系良好(r>0.999),低、中、高质控血浆样品的提取回收率为80.30%~96.45%。日内、日间精密度、准确度和稳定性均符合生物样品分析要求。结论建立的HPLC-FD定量分析方法快速、灵敏、准确,可用于人血浆中沙库巴曲、缬沙坦及其主要代谢产物LBQ-657的含量测定。
OBJECTIVE To establish an analytical method for simultaneous determination of sacubitril,valsartan and its main metabolite LBQ-657 in human plasmas by high performance liquid chromatography-fluorescence detection(HPLC-FD).METHODS Precipitation method was used to treat plasma samples via acetonitrile and SN-38 applied as internal standard.Agilent Eclipse SB-C_(18)(250 mm×4.6 mm,5μm)was used.The chromatographic separations were accomplished using multi-wavelength with a mobile phase composed of methanol and sodium dihydrogen phosphate buffer(pH 3.0)at ratio of 67∶33.The column temperature was maintained at 25℃ with the flow rate of 1.0 mL·min^(-1).The injection volume was 20μL.The multi-wavelength conditions were 0−6 min,λ_(EX)=380 nm,λ_(EM)=430 nm,for SN-38;6−9.4 min,λ_(EX)=255 nm,λ_(EM)=314 nm,for sacubitril;9−12 min,λ_(EX)=255 nm,λ_(EM)=374 nm,for valsartan;12−18 min,λ_(EX)=255 nm,λ_(EM)=314 nm,for LBQ-657.Sacubitril and valsartan sodium tablets(dose 100 mg)was administered to 5 patients and sampling after 1.5,3 h.HPLC-FD was used to detect concentration of sacubitril,valsartan and its main metabolite LBQ-657 in human plasma.RESULTS The calibration curves were linear over the range of 0.1-20μg·mL^(-1)(r>0.999).The extraction recovery of sacubitril,valsartan and LBQ-657 were found from 80.30% to 96.45%.According to the intra-and inter-day precisions,accuracy and stability,demonstrating that the method was confirmed to be accurate,precise and stable.CONCLUSION The developed method is successfully applied for determination of sacubitril,valsartan and its main metabolite LBQ-657 in human plasma.
作者
马妍妮
严宁
陈国霆
张辉
张文萍
MA Yanni;YAN Ning;CHEN Guoting;ZHANG Hui;ZHANG Wenping(Department of Pharmacy,General Hospital of Ningxia Medical University,Yinchuan 750004,China;Coronary Care Unit,General Hospital of Ningxia Medical University,Yinchuan 750004,China)
出处
《中国现代应用药学》
CAS
CSCD
北大核心
2022年第5期622-627,共6页
Chinese Journal of Modern Applied Pharmacy
基金
宁夏自然科学基金项目(2020AAC03363,2018AAC03140)。
