AC-012 苯乙胺手性方法.doc

Document Name: HPLC method for Phenethylamine Chiral purity determination Document NO: AC-012 Page : PAGE 1 of NUMPAGES 3 Effective Data: Jun 22,2016 Version Number: .00 Document Name: HPLC method for Phenethylamine chiral purity determination Revision History: Version Description of Change Date Author .00 New release Jun 22,2016 Feifei Hou Signatures: Author : Reviewed By: Date： Date： 1.Introduction: HPLC method for Phenethylamine chiral purity 2. Apparatus: 1) Standard laboratory equipment 2) Suitable HPLC chromatograph with UV detector or equipment equivalent. 3) Column: CHIRALCEL OD-H 4.6*250mm 5μm 3. Reagents: 1) n-hexane (HEX): HPLC grade 2) 2-Aminoethanlo（ETA）: Analytical reagent 3）Diethylamine（DEA）: Analytical reagent 4) Isopropanol (IPA): HPLC grade 5）Ethanol（ETOH）：HPLC grade 4.Chromatographic parameters: Column: CHIRALCEL OD-H 4.6*250mm 5μm Mobile Phase 1000HEX+40IPA+40ETOH+0.5DEA+0.5ETA Flow: 1.0 mL/min Column Temperature 25℃ Mode Isocratic Needle Wash Solvent Diluents Detection UV at 220nm Injection volume: 5μL Run time 15min 5.Mobile Phase and Diluents preparation Mobile Phase 1000HEX+40IPA+40ETOH+0.5DEA+0.5ETA Transfer 1000ml HEX and 40ml IPA 40ml ETOH to a suitable container. Add 0.5ml ETA and 0.5ml DEA in it, mix well. Diluents: Mobile Phase 6.Sample and standard Solution Preparation : Resolution Standard Preparation: Accurately weigh about 30 mg of R-Phenethylamine and 30 mg of S-Phenethylamine dissolved and diluted it to 100 ml volumetric flask with diluent .shake and mix well.（0.3mg/ml） sample solution preparation: Weigh about 30 mg of test sample. dissolved and diluted it to 100ml volumetric flask with diluent .shake and mix well.（0.3mg/ml） 7.System Preparation: Equilibrate the system until a stable baseline is observed. 8.Recommended Injection Sequence: Minimum one injection of blank Standard solution injection O