title: Resonance Rayleigh scattering for detection of proteins in HPLC.
abstract: An HPLC-resonance Rayleigh scattering (RRS) (HPLC-RRS) detection system is described for separation and detection of proteins. This system is based on the modification of a commercial HPLC instrument involving the addition of a pump and a T-shaped interface, and a common fluorescence detector was used for detection. The detection principle is based on the change of RRS intensity of the ion-association complex formed from biebrich scarlet (BS) and protein. The RRS signal was detected at lambdaex=lambdaem=376 nm. The utility of the presented method was demonstrated by the separation and determination of four proteins involving cytochrome (Cyt-c), lysozyme (Lys), HSA, and gamma-globulin (gamma-Glo). An LOD of 0.2-1.0 microg/mL was reached and a linear range was found between peak area and concentration in the range of 0.20-3.0 microg/mL for Cyt-c, 0.25-2.5 microg/mL for Lys, 1.5-10 microg/mL for HSA, and 2.0-15 microg/mL for gamma-Glo, with linear regression coefficients all above 0.99. The method presented has been applied to determine HSA and gamma-Glo in human serum samples synchronously.
authors.affiliation: College of Chemistry and Chemical Engineering, Guangxi, Normal University, Guilin, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Lu
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Luo
authors.forename: Chengwei
authors.initials: C
authors.lastname: Liu
authors.forename: Shulin
authors.initials: S
authors.lastname: Zhao
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D045304:Cytochromes c / Q000032:analysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009113:Muramidase / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000032:analysis; D013057:Spectrum Analysis / Q000295:instrumentation / Q000379:methods; D013997:Time Factors; D005719:gamma-Globulins / Q000032:analysis
chemical_list: D011506:Proteins; D012709:Serum Albumin; D005719:gamma-Globulins; D045304:Cytochromes c; D009113:Muramidase