The application discloses a preparation method of a
human insulin purity standard substance, which comprises the following steps of
raw material selection, qualitative identification, standard substance uniformity,
stability test, value determination and uncertainty evaluation. The application uses commercially available high-purity
human insulin as a
raw material, and after purity analysis, qualitative identification, uniformity test and
stability test,
isotope-labeled Val, Leu and Phe are used as internal standards, and liquid
chromatography-
isotope dilution mass spectrometry is used for value determination, so that the interference and influence of possible
isoleucine substitution
leucine impurities, isomer impurities or other molecular weight
protein impurities on the value determination result can be effectively excluded, and the value determination result is more accurate and reliable. The purity standard substance prepared by the application can be used for
serum insulin standard substance quality value tracing,
quality control of
insulin measurement results in in-vitro diagnosis, related method confirmation and evaluation, and evaluation of
repeatability,
linearity and cross
contamination rate of a closed light-emitting immune analyzer.