| In recent years, with the development of detection technology and sharply increased demand in relevant fields, serum estradiol and four purine nucleoside substances have become the new indicators in clinical laboratory, this paper developed serum estradiol and four kinds of purine nucleoside reference materials to ensure standardization of the measurement.In serum estradiol matrix reference materials development process, certification17β-estradiol by liquid chromatography-isotope dilution-mass spectrometry. Using electrospray ionization triple quadruple multiple reaction monitoring (MRM), achieving the best analytical conditions by optimizing chromatography and mass parameters, the pretreatment of serum samples is optimized, including comparison of different estradiol internal standard, the desired pH of serum, adding extractant, leaching conditions and equilibration time, to achieve the best extraction effect of estradiol and establish a suitable sample preparation route. It also tests the homogeneity, stability, evaluating the uncertainty and conducting method validation (precision, accuracy, linearity, limit of detection, etc).The results of the analysis are in the range of equivalent compared with2013IFCC RELA (JCTLM) sample and European Commission CRM. The measurement results traced to a recognized standard substance JCTLM to ensure that the standard substance traceability and accurate value to complete the development of serum estradiol matrix reference materials.Combining the related technology research abroad, the qualitative analysis of the four purine nucleoside samples was carried by infrared spectroscopy and mass spectrometry; the quantitative analysis of each reference material was evaluated by mass balance and nuclear magnetic resonance spectroscopy by optimizing the experimental conditions while completing measured the moisture, ash and inorganic ion content; the homogeneity, stability study and uncertainty were also detected. The results show that the four purine nucleoside reference materials have high purity and good homogeneity, and these samples are stable for a year at least, the purity of guanosine reference material was90.89%with the uncertainty of0.56%(k=2), the purity of2’-deoxyguanosine reference material was94.51%with the uncertainty of0.34%(k=2), the purity of8-oxo-guanosine reference material was94.22%with the uncertainty of0.34%(k=2), the purity of8-oxo-2’-deoxyguanosine reference material was91.02%with the uncertainty of0.28%(k=2). The development of the serum estradiol matrix reference material filled the domestic blank and the four purine nucleoside purity reference materials filled the international blank; such reference materials can be used for the determination of relative materials, the calibration of analysis instruments, analysis confirmed and the quality control for related departments and laboratories. |
