The Application Of Multi-way Calibration Method In Complex Systems Of Traditional Chinese Medicine And Proteins

Chemometrics is the main branch of analytical chemistry since it was born in1970s. It has profound great implications on the development of analytical chemistryeven for the whole area of chemistry through a rather short history. As an importantsection of chemometrics, multi-way calibrations have become a powerful tool formany scientists. And three-way calibration methodologies have the second-orderadvantage which quantifies analytes even in the presence of unknown interference, orphysical and chemical separation is enhanced or replaced by mathematical separation.In this paper, three-dimensional fluorescence spectram and second-order calibrationmethods were applied to research in the following aspects: traditional chinesemedicine and proteins. And it is organized into two parts and the main contents are asfollowing:Part Ⅰ: The quantitative research about the chinese medicine active substances inhuman serum and rheumatism wine (Chapter2to Chapter3)Curcuma longa L is a kind of traditional chinese medicine as early as the yearshave documented in the Tang dynasty, chemical research show that curcumin (CUR)and bisdemethoxycurcumin (BUR) are the main composition of Curcuma longa L.They have various kinds of biological activities and pharmaceutical value s, so it ismeaningful to have a quantitative research about them in human serum. In this chapter,a method based on three-dimensional fluorescence combined with alternating trilineardecomposition (ATLD) algorithm is established to determine the CUR and BUR inhuman serum directly even in the presence of interference, without complicatedpretreatment procedure and this method has a good performance.Scopoletin is one of the main active components in Feng Liaoxing rheumatismwine (FLX Wine) and the content of scopoletin is the standard to assess quality ofFLX Wine. It has the ability to cure the rheumatism and inflammation. So it isnecessary to detect the content of scopoletin in FLX Wine. Unfortunately，it isdifficult to achieve that by common HPLC method due to the fact that the matrix ofrheumatism wine is pretty complicated. In this chapter, a simple and fast method istaken to detect scopoletin based on three-dimensional fluorescence combined withchemometrics. The result is test by HPLC method and both of them are consistentwith each other.PartⅡ: The application of multi-way calibration in the interaction of epicatechin with protein molecules and the denaturizing process of HSA induced by guanidinehydrochloride (GdHCl)(Chapter4to Chapter5)The interaction between bovine serum albumin (BSA) and epicatechin wasstudied using fluorescence quenching titrations combined with trilineardecomposition method and excitation-emission matrix (EEM) fluorescence. Theresolved spectra are highly similar with the actual ones which indicated that theresolved results are reliable. The relevant parameters of the binding process areobtained by quantifying each substance in the complicated mixtures in situ. Thequenching is static quenching, epicatechin have a weak interaction with BSA and thebinding site is one. The total concentration and the free concentration of quenchershave different effect on the system. The results demonstrated that the method exploitin this article is a useful tool to investigate complex interactions, avoidingcomplicated pretreatment and simplify experimental procedure.Human serum albumin (HSA) is a common kind of globular proteins of thehuman circulatory system and of great importance in biochemisry and genetics. It hasa complex spatial structure with three domains and six subdomains. In this chapter,the unfolding of HSA induced by GdHCl was studied by principal component analysis(PCA) and three-dimensional fluorescence spectra combined with algorithmcombination methodology (ACM). An intermediate has been found and unfoldingGdHCl-induced HSA probably involved two stages. This conclusion is consistent withtraditional ‘fluorescence phase diagram’ approach and give a detailed researchcompared with the former.