Study On Separation And Analysis Of Several Active Components By High Performance Capillary Electrophoresis

Capillary electrophoresis is a separation technology combined electrophoresis with chromatography theory, which is an important progress in scientific analysis after HPLC. HPCE has the advantages of high efficiency, fast, micro sampling, various separation modes, extensive samples, economic, clean and high degree of automation. It is widely used in pharmaceutical analysis, food monitoring, environment, life sciences and other fields. In this paper, high performance capillary electrophoresis chiral separation mode selection and application of chiral selectors were briefly reviewed in recent years.And on this basis, four simultaneous separation and analysis method of biological active substances has been set up. The specific contents are as follows:1. A new capillary zone electrophoresis method using mannitol and methanol as organic additives for separation and determination of coptisine, berberine, epiberberine, palmatine and jatrorrhizine has been developed. The five kinds of alkaloids compounds could be effectively separated and determined within12min in a running buffer (pH8.05) consisting of50.0mmol/L mannitol,40.0mmol/L Na2HP04-NaH2PO4and25.0%(v/v) methanol with the applied voltage of22kV, capillary temperature of25℃, detector wavelength of230nm, and injection of5s at0.5psi. The linear range for the determination of coptisine, berberine, epiberberine, palmatine and jatrorrhizine were1.5～80.0,2.0～90.0,1.0～85.0,1.0～100.0and1.0～90.0μg/mL, respectively. The proposed method has been successfully used for determination of the five constituents in different types of coptis chinensis with the relative standard deviation of below5.0%and recovery ranged between95.3%and105.4%.2. A new capillary zone electrophoresis method for separation and determination of catechin (C), epicatechin (EC), gallocatechin gallate (GCG), epigallocatechin gallate (EGCG) and epigallocatechin (EGC) has been developed. The five kinds of polyphenolic compounds could be effectively separated and determined within13mins in a running buffer (pH8.68) consisting of12mmol/L Na2B4O7,60mmol/L NaH2PO4,4.0mmol/L β-CD and15.0%(v/v) ethylene glycol with the applied voltage of22kV, capillary temperature of25℃, detector wavelength of200nm, and injection of5s at 0.5psi. The linear range for the determination of C, EC, GCG, EGCG and EGC were1.0～110.0,2.0～90.0,2.0～90.0,1.0～100.0and1.0～90.0μg/mL, respectively. The proposed method has been successfully used for determination of the five constituents in different tea samples with the relative standard deviation of below5.0%and recovery ranged between96.1%and104.7%.3. A new RF-MEKC method was developed for the quantitave determination of praeruptorin A, praeruptorin E and praeruptorin C has been developed. The three compounds could be effectively separated and determined within15mins in a running buffer (pH2.12) consisting of10mmol/L NaH2PO4,140mmol/L SDS and40.0%(v/v) acetonitrile with the applied voltage of-20kV, capillary temperature of25℃, detector wavelength of200nm, and injection of5s at0.5psi. The linear range for the determination of praeruptorin A, praeruptorin E and praeruptorin C were2.0～95.0,1.0～90.0and1.0～90.0μg/mL, respectively. The proposed method has been successfully used for determination of the three constituents in different samples with the relative standard deviation of below5.0%and recovery ranged between95.4%and105.3%.4. A new RF-MEKC method was developed for the quantitave determination of Thr, Met, His, Phe, Val, Leu, Trp, Arg and Lys has been developed. The nine compounds could be effectively separated and determined within10mins in a running buffer (pH9.50) consisting of30mmol/L Na2B4O7,30mmol/L NaH2PO4,0.4mmol/L CTAB and2.8%(v/v) methanol with the applied voltage of-20kV, capillary temperature of25℃, detector wavelength of190nm, and injection of5s at0.5psi. The proposed method has been successfully used for determination of the nine common amino acids in Amino Acid Compound Injection(18) with the relative standard deviation of below5.0%and recovery ranged between95.2%and104.4%.