Study On The Method Of High-performance Capillary Electrophoresis And The Application In Traditional Chinese Medicine Analysis

The advantages of high performance capillary electrophoresis (HPCE) such as robustness, simplicity, low operational cost, more separation modes and better repellence make it an attractive alternative. The capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MECC) were applied to separation compounds with similar configuration and Traditional Chinese Medicine(TCM) analysis in this thesis. This investigation will be very useful for expanding the application of CZE, MECC, capillary partial filling technique (CPF) and related methods of chemometrics and developing analytical method for medicine. The thesis consists of seven chapters and main content as follows:1. HPCE was applied to separate the compounds with similar configuration. (1) The investigation about the separation of six benzoic acid compounds was carried out by MECC. Orthogonal array design for separation experiments was used to find the optimum conditions on the basis of resolution and analytical time. (2) Capillary partial filling -micellar electrokinetic capillary chromatography (CPF-MECC) was applied to separate six isomers of dinitrotoluene (DNT). The influential factors on the separation were investigated by orthogonal array design experiments. The result of this kind of separation was compared with the one by traditional MECC.2. To establish accurate methods for determination of effective components in Huaijiao pill and Sanhuang tablets by CZE, and in Zuguangsan by MECC:The quercetinum as internal standard substance, the CZE was established for simultaneous determination of genistein, rutin, baicalin and gallic acid in Huaijiao pills. An uncoated silica capillary was used with the running buffer solution of 15 mmol/L borate buffer solution (pH 9.0)-5 mmol/L sodium dihydrogen phosphate solution. The detection wavelength was 214 nm and voltage 18 kV. The four analytes were well separated within 12 min. The CZE was applied to the determination of baicalin in Sanhuang tablets according to experimental condition as follows: 40 mmol/L borax (pH8.95),λ=277nm, and voltage 18 kV.When benzoic acid and salicylic acid were determined synchronously in Zuguangsan , the MECC is applied with vitamin C as internal standard substance under the optimized condition: the running buffer solution of 20 mmol/L borate (pH9.8)-30 mmol/L SDS -4%methanol,λ=250 nm, and voltage 16 kV.In the above methods, all of the factors which affect the separation of the effective components in the electrophoresis system were investigated respectively. The methods were simple and quick. The results of determination were exact and credible. So these methods can be used to control the quality of TCMs.3. Considering the peaks of benzoic acid and salicylic acid are overlapped by CZE, partial least squares (PLS) in chemometrics was used to quantify benzoic acid and salicylic acid in Zuguangsan by CZE. Then the result is compared with the one by MECC. The two results are uniform. So the chemometric method can be able to separate the overlapping peak by HPCE in medicine analysis.