Fructus Aurantii Immaturus (枳实)-课件（PPT·精·选）.ppt

Agenda 8: Assay 1. Fructus Aurantii Immaturus (枳實) Determination of Synephrine in Fructus Aurantii Immaturus by HPLC Synephrine are used as the markers for the quality control of Fructus Aurantii Immaturus in . (2005) Same markers are used as TLC and fingerprint Synephrine Molecular weight: 167 Chemical formula: C 9 H 13 NO 2 NHCH 3 OH HO A U 0 . 0 0 0 . 0 2 0 . 0 4 0 . 0 6 0 . 0 8 0 . 1 0 0 . 1 2 0 . 1 4 n m 2 0 0 . 0 0 2 2 0 . 0 0 2 4 0 . 0 0 2 6 0 . 0 0 2 8 0 . 0 0 3 0 0 . 0 0 3 2 0 . 0 0 3 4 0 . 0 0 3 6 0 . 0 0 3 8 0 . 0 0 1 9 3 . 9 2 2 4 . 3 2 7 5 . 0 3 2 8 . 4 3 7 1 . 2 Selection of detection wavelength UV 224 nm ( Synephrine ) are chosen as detection wavelengths UV spectrum of Synephrine Selection of mobile phase (224 nm) Water – ACN ( 75 : 25 ) Isocratic Synephrine 30 32 68 32680 ACN Water Time Same as fingerprint condition Water (%SDS, %H 3 PO 3)– ACN ( 68 : 32 ) Isocratic Synephrine The retention times of Synephrine are very different due to their polarity and structures . Synephrine + SDS = stability NHCH 3 OH HO SDS-SO 3 - + Selection of extraction solvent 50% M ethanol is chosen as the extraction solvent. ZS sample ( g) + 25 mL solvent sonicate 30 min x 2 HPLC analysis 75% MeOH MeOH 2132940 50%MeOH Synephrine 272807 CH 2 Cl 2 627431 EtOH 680923 H 2O 1634510 25%MeOH 1975483 75%MeOH 1519732 MeOH Peak area Solvent One-way ANOVA (n =3) 50% MeOH CH 2 Cl 2 EtOH H 2O 25% MeOH 202578 1 202578 1 Selection of extraction method Sonicate 30 min twice is chosen as the extraction method. Peak area Extraction method Synephrine 1852777 Soxhlet extraction 2079484 Reflux 2118892 Sonicate One-way ANOVA (n =3) ZS sample ( g) + MeOH HPLC analysis Reflux 2 hr Soxhlet Extraction 4 hr Sonicate 30 min x 2 HPLC condition for determination of andrographolide and dehydroandrographolide in Herba Andrographidis Column : Injection volume: Detector: Detector wavelength: Flow rate: Mobile phase: C 18 HPLC column, A