分析测试学报2025，Vol.44Issue(5)：811-821,11.DOI:10.12452/j.fxcsxb.240913396
柱前衍生/高效液相色谱-荧光检测法测定胰腺癌PANC-1和CFPAC-1细胞荷瘤小鼠中13种硫酸乙酰肝素二糖
Determination of 13 Heparan Sulfate Disaccharides in PANC-1 and CFPAC-1 Cells Tumor Bearing Mice with Pancreatic Cancer by Precolumn Derivatization/High Performance Liquid Chromatography and Fluorescence Detection
摘要
Abstract
A precolumn derivatization/high performance liquid chromatography-fluorescence detec-tion(HPLC-FLD)method for the determination of 13 heparan sulfate(HS)disaccharides in PANC-1 and CFPAC-1 cells tumor bearing mice with pancreatic cancer was established.HS disaccharides were obtained from pancreatic cancer samples by defatting and protease enzymatic hydrolysis,and then reacted with 2-aminoacridone(AMAC)derived reagents.The separation was performed by Ther-mo Scientific HYRERSIL ODS-2 C18 column,and the mobile phase gradient elution was performed with methanol-60 mmol/L ammonium acetate(pH 5.6)at excitation wavelength of 425 nm and emis-sion wavelength of 520 nm.The results showed that the 13 HS disaccharides had a good linear rela-tionship within the corresponding mass concentration ranges(r2≥0.999 0),with detection limits(LODs)of 0.012-0.313 μg/mL and the quantitation limits(LOQs)of 0.117-1.250 μg/mL.The rel-ative standard deviations(RSDs)of precision ranged from 0.040%to 0.63%,the RSD of intra-day stability ranged from 0.34%to 3.6%,the RSD of inter-day stability ranged from 1.8%to 6.4%.The recoveries ranged from 80.7%-109%,and RSD was 0.50%-17%.9 HS disaccharides were detected in the tumor tissue of the two types of pancreatic cancer cells,among which ΔUA-GlcN(ⅣH)ac-counted for the highest proportion,with concentrations of 28 627.558 nmol/g and 66 938.786 nmol/g,respectively.ΔUA-GlcNAc(ⅣA)and ΔUA-GlcN,6S(ⅡH)took the second place,and the results of molecular docking showed that ⅣH had the strongest binding force with heparin-binding epidermal growth factor(HB-EGF)and ⅡH had the strongest binding force with fibroblast growth fac-tor-2(FGF-2).The characteristics of HS sulfated sequence and its interaction with HB-EGF and FGF-2 regulate the occurrence and development of pancreatic cancer.The findings of this study may provide valuable insights for the early treatment and diagnosis of pancreatic cancer.关键词
胰腺癌/硫酸乙酰肝素/高效液相色谱-荧光检测法/柱前衍生Key words
pancreatic cancer/heparan sulfate(HS)/HPLC-FLD/precolumn derivatization分类
化学引用本文复制引用
张蕾,王思亚,张运涛,任强..柱前衍生/高效液相色谱-荧光检测法测定胰腺癌PANC-1和CFPAC-1细胞荷瘤小鼠中13种硫酸乙酰肝素二糖[J].分析测试学报,2025,44(5):811-821,11.基金项目
山东省自然科学面上项目(ZR2022MH176) （ZR2022MH176）
济宁医学院高层次科研项目(JYGC2021KJ012) （JYGC2021KJ012）
