Study On The Determination Of Some Pharmaceuticals By Resonance Rayleigh Scattering And High Performance Liquid Chromatography

Spectrophotometry is a classic, rapid, simple and convenient analytical technology, which is widely used to analyze pharmaceuticals. In recent years, Resonance Rayleigh scattering (RRS) is a new analytical technology which is provided with high sensitivity, cheapness instrument, simple operation and rapid analysis speed. At present, the RRS method has brought to much more attention and interesting for the researchers in determining pharmaceuticals and other fields. It has been quickly developed and has been applied in the determination of biomacromolecules, trace metal ions and nonmetals as well as organic matters.High Performance Liquid Chromatography (HPLC) is a new separation technology based on classic liquid chromatography and gas chromatography in 20c 60's. It adopts new style high pressure pump, high sensitivity detector, high efficient performance particulate solid phase. HPLC has a fast development speed, and online analysis can be achieved by connecting with other analytical instruments. Up to now, HPLC has been widely used in environment science, agriculture, chemistry, material, biology, medicine pharmaceutics and so forth.In this dissertation, some organic matter such as caffeic acid (CA) interaction with Pb(â…¡), Ag (?)-Phenanthroline have been studied by the RRS technology, and we have also researched the RRS phenomenon between two type of acid mucopolysaccharides (dextran sulfate sodium and heparin ) and Ag(â… )-Phenanthroline. In addition, the determination chlorogenic acid and caffeic (?)id in Kang Yan Ling Stomatitis Spray has been discussed by reverse phase high performance liquid chromatography(RP-HPLC). The main contents are as follows:1. Resonance Rayleigh scattering and resonance nonlinear scattering spectroscopy of Pb(â…¡)-caffeic acid chelate complex and its analytical applicationIn the medium of 5,5-diethylbarbituric acid sodium and sodium acetate buffer solution (pH 6.80), the complex of caffeic acid and Pb(â…¡) will significantly lead to the enhancement of the resonance Rayleigh scattering (RRS) intensity, second-order scattering (SOS) intensity and frequence-double scattering (FDS) intensity. The maximum RRS peak was at 470 nm, and another small scattering peak was at 320 nm. The maximum SOS peak was at 560nm, and the maximum FDS peak was at 445nm. The three scattering intensities are directly proportional to the concentration of caffeic acid in range of 2.0×10^-6 mol·L^-1-4.0×10^-5mol·L^-1 for RRS method and SOS method, 5.0×10^-7mol·L^-1 -1.5×10^-5mol·L^-1 for FDS method. The detection limit of caffeic acid is 1.62×10^-8mol·L^-1 for RRS method, 7.90×l0^-8mol·L^-1 for SOS method, 3.45×l0^-7 mol·L^-1 for FDS method. The optimum conditions and coexistent materials were discussed. The results suggest that the novel sensitive, simple and quick scattering methods for the determination of caffeic acid have been established.2. Determination of caffeic acid with Ag (â… )-Phenanthroline -caffeic acid in system by Resonance Rayleigh Scattering methodIn the medium of pH 5.40 HAc-NaAc buffer solution,the complex of caffeic acid(CA) and Ag (â… )-Phenanthroline will significantly lead to the enhancement of the resonance Rayleigh scattering (RRS) intensity. The maximum RRS peak was at 386 nm. The scattering intensity are directly proportional to the concentration of caffeic acid in range of 2.0×l0^-7mol·L^-1-1.0×10^-4mol·L^-1 for RRS method. The detection limit of caffeic acid is 1.02×10^-7 mol·L^-1. The optimum conditions and coexistent materials were discussed. The novel sensitive, simple and quick scattering method for the determination of caffeic acid has been found.3. Determination of caffeic acid with Ag (â… )-Phenanthroline-dextran sulfate sodium in system by resonance Rayleigh scattering methodA new Rayleigh scattering method for determination dextran sulfate sodium is presented.In pH 3.40 HAc-NaAc buffer solution, it is found that when [Ag(Phen)₂]⁺ and dextran sulfate sodium react through electrostatic force to form the complex [Ag(Phen)₂]⁺-dextran sulfale sodium quickly. the RRS intensity of the system will be significantly enhanced. The maximum RRS peak was at 400 nm. The scattering intensity is directly proportional to the concentration of dextran sulfate sodium in range of 0.04-2.0μg·mL^-1,and the detection limit of dextran sulfate sodium is 2.8 ng·mL^-1. The optimum conditions and coexistent materials were discussed. This method can be used for the determination of dextran sulfate sodium in synthetical samples with satisfactory results.4.Determination of caffeic acid with Ag (â… )-Phenanthroline -heparin in system by resonance Rayleigh scattering methodA novel method for the determination of heparin(Hep) with Ag(â… )-Phenanthroline by resonance Rayleigh scattering has been proposed. In the medium of HAc-NaAc buffer solution (pH 5.40), the interaction of heparin with [Ag(Phen)₂]⁺ can be occurred which greatly enhanced the RRS intensity at 398nm. The scattering intensity is directly proportional to the concentration of heparin in the range of 0.04-6.0μg·mL^-1 and its detection limit is 20 ng·mL^-1.The optimum conditions and coexistent materials were discussed. The novel scattering method is satisfying when applied in the determination of heparin in actual sample.5. Determination chlorogenic acid and caffeic acid in Kang Yan Ling Stomatitis Spray by RP-HPLCA HPLC method was established for the determination of chlorogenic acid and caffeic acid in Kang Yan Ling Stomatitis Spray.This analysis was performed on a Shimadzu Shim -pack VP-ODS column with methanol and 0.012 mol·L^-1 phosphoric acid(27:73)as mobile phase. The temperature of the column was set at 30℃. The flow rate was 0.8 mL·min^-1 and the detection wavelength was 232 nm. Chlorogenic acid and caffeic acid in KangYan Ling Stomatitis Spray were highly separated and exactly quantified.The linear concentration range of chlorogenic acid was 0.51-91.80μg·mL^-1 with the recovery of 97.5% (RSD=1.0 %).The detection limit (S/N=3) was 0.10μg·mL¹, and the quantified detection (S/N=10) was 0.36μg·mL^-1. The linear concentration range of caffeic acid was 2.0-360.0μg·mL^-1 with the recovery of 100.4% (RSD=1.0%).The detection limit (S/N=3) was 0.40μg·mL^-1, and the quantified detection (S/N=10) was 1.40μg·mL^-1. This method is simple and convenient with the capability of high veracity and high reproducibility and can be used to control the quality of Kang Yan Ling Stomatitis Spray.