Hollow Fiber Cell Anthraquinones Active Ingredient Groups Capture And Phacoemulsification Ionic Liquid Micro-extraction Analysis

In the study, colorectal cancer cell HCT116was firstly seeded in hollowfiber as an active screening tool, then the hollow fiber with cells was inserted intoTCMs extract solution to screen and fish anthraquinones active ingredients. After that,anthraquinones bound with cells were desorbed with methanol and analyzed by HPLC.So a new bioactive ingredient screening method of hollow fiber cell fishing with highperformance liquid chromatography (HFCF-HPLC) has been developed andintroduced for the screening and fishing bioactive ingredients from traditional Chinesemedicines (TCMs). The surface properties of hollow fiber seeded HCT116cells, thenonspecific binding between active center in fiber and the target ingredients, the cellsurvival rate under different conditions before and after the screening, and therepeatability and stability of hollow fiber HCT116cell-fishing with HPLC wereanalyzed and investigated. On the basis, cell fishing factor of active ingredient wasdefined in the HFCF-HPLC. Some structures of anthraquinones active ingredientsscreened from TCMs were identified by the comparison to the retention times of thereference substances elucidated by mass spectrometry. Meanwhile, indometacin wasused as positive control. The test results demonstrate that HFCF-HPLC is a simple,fast, effective and reliable method for screening and analyzing the bioactiveingredients. It also has the potential to screen other bioactive ingredients from TCMs. An ultrasensitive method of ultrasound emulsifcation ionic liquidmicroextraction (UEILME) coupled with high performance liquid chromatography(HPLC) has been developed and introduced for the preconcentration and analysis offive anthraquinone compounds (aloe-emodin, rhein, emodin, chrysophanol, andphyscion) in traditional Chinese medicines and anthraquinone additives in cosmeticsamples. Several parameters affecting the extraction efficiency, such as the type andamount of extraction solvent, sample pH, ultrasound time and temperature,centrifugation speed and time, and ionic strength, were investigated and optimized.The most favorable results were obtained using60mg1-hexyl-3-methylimidazoliumhexafluorophosphate ([C6MIM][PF6]) as extraction solvent. The anthraquinones wereextracted from the aqueous solution (pH2.0) by ultrasound at40℃for7min andthen centrifuged at2500rpm for6min. Under optimal conditions, the good linearitiesof the five anthraquinone compounds were obtained with correlation coefficients>0.99. The limits of detection (LODs) and the limits of quantitation (LOQs) rangedfrom10to90g/mL and50to250g/mL, respectively. The relative standarddeviations (n=3) were less than9.8%. Moreover, the enrichment factors ranged from80to197folds. Compared with conventional dispersive liquid–liquid microextraction,the UEILME technique exhibited lower LODs and LOQs. The results demonstratedthat the UEILME coupled with HPLC is a simple, efficient, sensitive, andenvironment-friendly method for the extraction, concentration and analysis ofanthraquinone compounds.