• Bulletin of the Korean Chemical Society
• /
• v.34 no.12
• /
• pp.3817-3824
• /
• 2013

Various separation modes in HPLC, such as anion exchange (AE), reversed-phase (RP), and affinity (AF) chromatography were examined for the separation of selenium species in human blood serum and urine. While RP- and AE-HPLC were mainly used for the separation of small molecular selenium species, double column AF-HPLC achieved the separation of selenoproteins in blood serum efficiently. Further, the effluent of AF-HPLC was enzymatically hydrolyzed and then analyzed with RP HPLC for selenoamino acid study. The versatility of the hybrid technique makes the in-depth study of selenium species possible. For quantification, post column isotope dilution (ID) with $^{78}Se$ spike was performed. ORC ICP/MS (octapole reaction cell inductively coupled plasma/mass spectrometry) was used with 4 mL $min^{-1}$ Hydrogen as reaction gas. In urine sample, inorganic selenium and SeCys were identified. In blood serum, selenoproteins GPx, SelP and SeAlb were detected and quantified. The concentration for GPx, SelP and SeAlb was $22.8{\pm}3.4\;ng\;g^{-1}$, $45.2{\pm}1.7\;ng\;g^{-1}$, and $16.1{\pm}2.2\;ng\;g^{-1}$, respectively when $^{80}Se/^{78}Se$ was used. The sum of these selenoproteins ($84.1{\pm}4.4\;ng\;g^{-1}$) agrees well with the total selenium concentration measured with the ID method of $87.0{\pm}3.0\;ng\;g^{-1}$. Enzymatic hydrolysis of each selenium proteins revealed that SeCys is the major amino acid for all three proteins and SeMet is contained in SeAlb only.

• Analytical Science and Technology
• /
• v.28 no.6
• /
• pp.417-424
• /
• 2015

In this study, the relationship between selenoprotein concentrations in blood and stomach cancer have been searched for Korean. The concentration of each selenoprotein in blood serum was analyzed and the correlation between the concentration and stomach cancer was studied to find a potential for using Selenium as a biomarker. In concentration determination, a simple calibration curve method was used with the monitoring of m/z 78 without the use of solid phase extraction. This is a lot more simple than the method using SPE with post column isotope dilution. The result obtained from the analysis of CRM BCR-637, 72.20±3.35 ng·g⁻¹, showed similar value of reference value (81±7 ng·g⁻¹). The total concentration of Se for the controlled group, cardiovascular patients group, was 105.70±21.20 ng·g⁻¹. This value was the same as normal healthy person reported earlier. Each selenoprotein concentration of GPx, SelP and SeAlb was 26.12±7.84, 65.15±14.50, 14.43±6.99 ng·g⁻¹, respectively. The distribution of each selenoprotein was 24.7%, 61.6%, and 13.7%, which was similar to the normal person. The result of stomach cancer patients, the total concentration of Se was 76.11±28.12 ng·g⁻¹ and each concentration of GPx, SelP and SeAlb was 15.41±9.01, 50.83±17.91, and 9.87±5.21 ng·g⁻¹, respectively. The total and each selenoprotein concentration level showed significant decrease for the stomach cancer patients. The level of decrease was 41.0% for GPx, 22.0% for SelP, and 31.6% for SeAlb. However, the distribution of each selenoprotein was not much different. Either total Selenium or each selenoprotein could be used as a possible index for the diagnosis of cancer. However, in age group study, it is shown that young age group (30's-40's) did not show much difference.

• Analytical Science and Technology
• /
• v.27 no.2
• /
• pp.92-99
• /
• 2014

Interferences were removed using anion exchange solid phase extraction (AE SPE) in quantification of selenoproteins 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 $94.3{\pm}2.3ngg^{-1}$ 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 $85.4{\pm}3.4ngg^{-1}$, which agreed well with the reference value of $81{\pm}7ngg^{-1}$. The pooled concentration of GPx, SelP, and SeAlb from healthy Koreans (n=20) was $12.1{\pm}1.4ngg^{-1}$, $57.2{\pm}2.0ngg^{-1}$, and $20.0{\pm}1.9ngg^{-1}$, respectively. The sum of selenoproteins is $89.3ngg^{-1}$, which is about the same as the total selenium concentration of $94.3ngg^{-1}$. 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 $25.0ngg^{-1}$ to $12.1ngg^{-1}$. 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 Cl and Br that cause interference to GPx.

• Analytical Science and Technology
• /
• v.28 no.3
• /
• pp.196-203
• /
• 2015

Selenium species (inorganic selenium, selenoaminoacids, and selenoproteins) were analyzed using anion exchange and affinity chromatography, which were connected to ICP/MS for the blood serum of sows fed by seleniumfortified feed. The Anion Exchange PRP X-100 column was used for the analysis of inorganic selenium (Se⁴⁺ and Se⁶⁺) and selenoaminoacids. The HEP column was used to separate SelP from GPx+SeAlb in selenoproteins. A quantitative analysis was performed using the post-column isotope dilution technique. The lactating sows were divided into three groups and fed by selenium fortified feed (organic 0.3 mg/kg, 0.6 mg/kg and inorganic 0.6 mg/kg) for four weeks. The test groups showed increases in selenoaminoacids compared with the control group, except the inorganic feed group. There was no significant difference between the organic feed groups. All test groups showed increases in selenoproteins. In particular, SelP showed a large increase that was 1.5 times higher than the other proteins.

• Journal of the Korean Chemical Society
• /
• v.63 no.6
• /
• pp.420-426
• /
• 2019

Selenoproteins,in Korean blood serum, GPx, SelP, and SeAlb were separated and determined with the use of HPLC-ICP/MS. Deuterium was used as a collision gas and affinity column with ammonium formate was used as an eluting solvent for the accurate quantitation of selenoproteins in human blood serum. Certified reference material BCR 639 (133±12 ng g^-1) was tested for the accuracy and the result was satisfactory 130±6 ng g^-1. Blood serum for the rectal cancer and controlled groups were collected and analyzed to give 84±27 ng g^-1, and 119±28 ng g^-1, respectively. The difference was statistically obvious when t-test was performed (t_cal 4.93 > t_95% 2.04). The decrease for cancer group was more obvious for female and aged group. The distributions of three selenoproteins were similar with each other, which means rectal cancer group did not show any specificity for any selenoproteins. As cancer developed, GPx showed a slight decrease but not obvious while the total concentration was increasing particularly at the second stage of cancer.

• Journal of Pharmaceutical Investigation
• /
• v.6 no.2
• /
• pp.101-110
• /
• 1976

1. In the rabbit and the dog, the blood pressure response to water extract and methanol extract obtained from Atractylodes rhizoma alb'a was investigated. 2. Water extract and methanol extract, when administered into the rabbit and the dog by the route of vein, produced fall of the blood pressure. 3. The depressor response of the rabbit to water extract and methanol extract was not affected by $Avicel{\circledR}$, propranolol and atropine. 4. The depressor response by water extract and methanol extract in the rabbit was not affected by guanethidine, but water extract and methanol extract produced elevation of blood pressure in this rabbit. 5. Pretreatment of rabbit with chlorisendamine or phenoxybenzamine weakened the depressor response to water extract and methanol extract, and the both extracts produced secondary elevation of blood pressure in this rabbit. 6. The pressor response of the chlorisondamine-treated rabbit to water extract and methanol extract was not affected by atropine. 7. Water extract decreased the pressor action of tyramine and depressor action of pilocarpine and isoproterenol, but did not affect the blood pressure response of nor einephrine, angiotensin and dimethylpehnyl piperazinium iodide(DMPP).