고체상 추출과 친화 크로마토 그라피-유도결합 플라즈마 질량분석법을 이용한 한국인 혈청에서의 셀레노 단백질 정량

Abstract

Interferences were removed using anion exchange solid phase extraction (AE SPE) in quantification of seleno proteins in korean human blood serum with affinity high performance liquid chromatography (AF HPLC)-inductively coupled plasma/mass spectrometry (ICP/MS). The average selenium level obtained for healthy koreans was 94.3 ± 2.3 ng g �1 using isotope dilution method. AE SPE was coupled to AF column to separate 3 selenoproteins, glutathione peroxidase (GPx), selenoprotein (SelP), and selenoalbumin (SeAlb). Post column isotope dilution was employed to quantify the proteins. The certified reference material of human blood serum, BCR-637, was analyzed to give total selenoprotein concentration of 85.4 ± 3.4 ng g �1 , which agreed well with the reference value of 81 ± 7 ng g �1 . The pooled concentration of GPx, SelP, and SeAlb from healthy koreans (n=20) was 12.1 ± 1.4 ng g �1 , 57.2 ± 2.0 ng g �1 , and 20.0 ± 1.9 ng g �1 , respectively. The sum of selenoproteins is 89.3 ng g �1 , which is about the same as the total selenium concentration of 94.3 ng g �1 . The fact suggests that selenium in blood serum is mostly consisted of selenoproteins. After the removal of interference, GPx showed a significant decrease (more than 50%) from 25.0 ng g �1 to 12.1 ng g �1 . It was identified that the interference in blood serum was mostly from GPx and the use of AE SPE was proven to be efficient in eliminating Cland Brthat cause interference to GPx. 요약 : 본 연구에서는 음이온 고체상 추출법 (AE SPE; anion exchange solid phase extraction)을 사용하 여 간섭요인을 제거한 후, 친화 크로마토그래피 AF HPLC; affinity high performance liquid chromatography) 와 유도결합 플라스마 질량분석법 (ICP/MS; inductively coupled plasma/mass spectrometry)을 사용하여 한