S detector (Thermo Electron, San Jose, CA) incorporated with heated electrospray ionization (H-ESI) interfaces. A Gemini C18 column (5062.0 mm i.d., 3 mm; Phenomenex, Torrance, CA) was used for separation of theaflavins and their potential metabolites at a flow rate of 0.2 mL/min. The column was eluted with 100 solvent A (H2O with 0.1 formic acid) for 3 min, followed bylinear increases in B (acetonitrile with 0.1 formic acid) to 70 from 3 to 48 min and to 100 B from 48 to 49 min, and then with 100 B from 49 to 54 min. The column was re-equilibrated with 100 A for 5 min. A Gemini C18 column (15063.0 mm i.d., 5 mm; Phenomenex, Torrance, CA) was used for separation of phenolic acids and their potential metabolites at a flow rate of 0.3 mL/min. The column was eluted with 100 solvent A (H2O with 0.1 formic acid) for 5 min, followed by linear increases in B (acetonitrile with 0.1 formic acid) to 100 from 5 to 15 min, and then with 100 B from 15 to 20 min. The column was reequilibrated with 100 A for 5 min. The LC eluent was introduced into the H-ESI interface. The negative ion polarity mode was set for the H-ESI source with the voltage on the H-ESI interface maintained at approximately 4 kV. Nitrogen gas was used as the sheath gas and auxiliary gas. To detect the theaflavins and their metabolites, optimized source parameters, including ESI capillary temperature (300uC), capillary voltage (?0 V), ion spray voltage (3.6 kV), sheath gas flow rate (30 units), auxiliary gas flow rate (5 units), and tube lens (?20 V), were tuned using authentic TFDG. To detect the phenolic acids and their metabolites, optimized source parameters were tuned using authentic gallic acid. These parameters include ESI capillary temperature (300uC), capillary voltage (?0 V), ion spray voltage (3.6 kV), sheath gas flow rate (35 units), auxiliary gas flow rate (15 units), and tube lens (?0 V). The collision-induced dissociation (CID) for H-ESI was conducted with an isolation width of 2 Da and normalized collision energy of 35 for MS2 and MS3. Default automated gain control target ion values were used for MS, MS2, and MS3 analyses. The mass range was from 50 23727046 to 1000 m/z for detection TFs and their metabolites, from 50 to 400 m/z for detection phenolic acids and their metabolites. The mass resolution was 0.6 amu FWHM. Data acquisition was performed with Xcalibur version 2.1.0 (Thermo Electron, San Jose, CA).Author ContributionsConceived and designed the experiments: SS CJ SAI. Performed the experiments: HC SH JRG. Analyzed the data: HC SS. Contributed reagents/materials/analysis tools: SS NDG. Wrote the paper: SS HC CJ.
Liver diseases and injuries are important medical problem worldwide. Liver purchase Bexagliflozin transplantation is currently the most efficient therapy for liver failure and end-stage liver disease. However, it is Pentagastrin limited by the scarcity of donor, expensive medical 15755315 cost, surgical risk and requiring life-long immunosuppressant agents. The development and application of hepatocytes transplantation has been attempted to treat different forms of liver diseases [1,2,3]. It has minimal invasive procedures and fewer surgical complications compared to the orthotopic liver transplantation. Stem cell transplantation has also gained considerable attention recently. Stem cells have the potential to supportive tissue regeneration andto generate large amounts of donor cells ready for transplantation [4,5,6,7]. The induced pluripotent stem cells (iPS) are generated from differentiat.S detector (Thermo Electron, San Jose, CA) incorporated with heated electrospray ionization (H-ESI) interfaces. A Gemini C18 column (5062.0 mm i.d., 3 mm; Phenomenex, Torrance, CA) was used for separation of theaflavins and their potential metabolites at a flow rate of 0.2 mL/min. The column was eluted with 100 solvent A (H2O with 0.1 formic acid) for 3 min, followed bylinear increases in B (acetonitrile with 0.1 formic acid) to 70 from 3 to 48 min and to 100 B from 48 to 49 min, and then with 100 B from 49 to 54 min. The column was re-equilibrated with 100 A for 5 min. A Gemini C18 column (15063.0 mm i.d., 5 mm; Phenomenex, Torrance, CA) was used for separation of phenolic acids and their potential metabolites at a flow rate of 0.3 mL/min. The column was eluted with 100 solvent A (H2O with 0.1 formic acid) for 5 min, followed by linear increases in B (acetonitrile with 0.1 formic acid) to 100 from 5 to 15 min, and then with 100 B from 15 to 20 min. The column was reequilibrated with 100 A for 5 min. The LC eluent was introduced into the H-ESI interface. The negative ion polarity mode was set for the H-ESI source with the voltage on the H-ESI interface maintained at approximately 4 kV. Nitrogen gas was used as the sheath gas and auxiliary gas. To detect the theaflavins and their metabolites, optimized source parameters, including ESI capillary temperature (300uC), capillary voltage (?0 V), ion spray voltage (3.6 kV), sheath gas flow rate (30 units), auxiliary gas flow rate (5 units), and tube lens (?20 V), were tuned using authentic TFDG. To detect the phenolic acids and their metabolites, optimized source parameters were tuned using authentic gallic acid. These parameters include ESI capillary temperature (300uC), capillary voltage (?0 V), ion spray voltage (3.6 kV), sheath gas flow rate (35 units), auxiliary gas flow rate (15 units), and tube lens (?0 V). The collision-induced dissociation (CID) for H-ESI was conducted with an isolation width of 2 Da and normalized collision energy of 35 for MS2 and MS3. Default automated gain control target ion values were used for MS, MS2, and MS3 analyses. The mass range was from 50 23727046 to 1000 m/z for detection TFs and their metabolites, from 50 to 400 m/z for detection phenolic acids and their metabolites. The mass resolution was 0.6 amu FWHM. Data acquisition was performed with Xcalibur version 2.1.0 (Thermo Electron, San Jose, CA).Author ContributionsConceived and designed the experiments: SS CJ SAI. Performed the experiments: HC SH JRG. Analyzed the data: HC SS. Contributed reagents/materials/analysis tools: SS NDG. Wrote the paper: SS HC CJ.
Liver diseases and injuries are important medical problem worldwide. Liver transplantation is currently the most efficient therapy for liver failure and end-stage liver disease. However, it is limited by the scarcity of donor, expensive medical 15755315 cost, surgical risk and requiring life-long immunosuppressant agents. The development and application of hepatocytes transplantation has been attempted to treat different forms of liver diseases [1,2,3]. It has minimal invasive procedures and fewer surgical complications compared to the orthotopic liver transplantation. Stem cell transplantation has also gained considerable attention recently. Stem cells have the potential to supportive tissue regeneration andto generate large amounts of donor cells ready for transplantation [4,5,6,7]. The induced pluripotent stem cells (iPS) are generated from differentiat.