Application Studies Of HPLC With Chemiluminescence Detection For Food And Pharmaceutical Samples

Chemiluminescence(CL) is defined as the emission of light from an electronically excited species which produced during the course of a chemical reaction,and is observed when the electronically excited product or intermediate formed decays to the ground state with a photon emitted.There are many merits on CL such as high sensitivity,wide linear range,rapidity,simplicity and so on.Due to the power ability of separation,high performance liquid chromatography coupled with chemiluminescence has been widely applied in biological science,food science and environmental detection.The thesis consists of two parts.One is review related to the application of HPLC with post-column chemiluminescence detection to food and pharmaceutical samples. Emphasis is taken on the development in the recent years.The other is research reports which is composed of 4 components as follows:1) The determination of trans-resveratrol in grape by HPLC-Chemiluminescence detection;2) Determination of levamisole and mebendazole in milk powder by matrix solid-phase dispersion and HPLC;3) Rapid determination of tetracyclines antibiotics and chloramphenicol in pig liver by HPLC;4) Determination of tetrahydropalmatine and ferulic acid in Tianqi Tonging capsule by HPLC.1.The Determination of Trans-Resveratrol in Grape by HPLC and Chemiluminescence DetectionA novel chemiluminescence.determination of trans-resveratrol in grape by high performance liquid chromatographic(HPLC) separation with chemiluminescence detector was described.The signal produced by the chemiluminescent reaction(CL) between acidic potassium permanganate and sodium hyposulfite was greatly increased in presence oftrans-resveratrol.The linear ranges was 0.1～20μg/mL(r=0.9996) and the detection limit is 0.03μg/mL for trans-resveratrol.The method has been satisfactorily applied for the determination of trans-resveratrol in grapes.2.Determination of Levamisole and Mebendazole in Milk Powder by Matrix Solid-Phase Dispersion and High Performance Liquid ChromatographyThe analysis method on levamisole and mebendazole in milk powder was studied. Experiments were performed on Hypersil C₁₈ column(4.6 mm×l50 mm,5.0μm) and detected at 230 nm.The mobile phase was composed of methanol-potassium dihydrogen phosphate at a flow rate of 0.8 mL/min.The linear range of levamisole and mebendazole was 0.09～1.44μg/mL and 0.06～0.96μg/mL respectively with the detection limit of 0.09μg/mL and 0.06μg/mL.The average recoveries of levamisole and mebendazole was 69.00%～80.00%.The method is simple,rapid and accurate.3.Rapid Determination of Tetracyclines Antibiotics and Chloramphenicol in Pig Liver by Reversed-Phase High Performance Liquid ChromatographyThe analysis method on tetracyclines antibiotics(TCs) and chloramphenicol in pig liver were studied.Experiments were performed on Hypersil C₁₈ column(4.6 mm×150 mm,5.0μm) and detected at 268 nm and at room temperature.The mobile phase was composed of methanol-acetonitrile-oxalic acid at a flow rate of 1.0 mL/min.The linear range of the TCs and chloramphenicol was 0.04～1.0μg/mL while the detection limit was 0.02～0.06μg/mL.The average recoveries of them were 80.00%～116.0%and the coefficient were 1.30～5.24%.The method is rapid,simple,accurate and suitable for routine analysis.4.Determination of Tetrahydropalmatine and Ferulic Acid in Tianqi Tongjing Capsule by HPLCAn HPLC method is developed for the simultaneous separation and determination of tetrapalmatine and ferulic acid in Tianqi Tongjing capsule.Experiments were performed on Hypersil C₁₈ column(4.6 mm×150 mm,5.0μm) and detected at 288 nm The mobile phase was composed of methanol-potassium dihydrogen phosphate at a flow rate of 0.8 mL/min.The linear range of tetrahydropalmatine and ferulic acid was 0.13～1.29μg and 0.08～0.80μg respectively.The average recoveries of tetrahydropalmatine and ferulic acid were 97.96%(RSD=1.3%) and 98.47%(RSD=1.1%).The method is accurate,simple,effective,and reproductive.