- P-ISSN 1225-0163
- E-ISSN 2288-8985
본 연구에서는 HPLC-ICP-MS를 이용하여 여러 식품에 포함된 셀레늄의 화학종을 분리하고 정량하였다. 양이온 교환 크로마토그래피를 사용할 때에 무기셀레늄 화학종의 분리는 효율적이나 유기셀레늄화학종은 분리가 완전하지 못한 반면, 역상 이온쌍 크로마토그래피에서는 유기와 무기 셀레늄 화학종들을모두 잘 분리 할 수 있었다. C8 컬럼(SymmetryshieldTM RP8, 3.5 μm, 4.6 × 150 mm)을 이용하여 최적조건(이동상; 5% 메탄올, ion-pairing reagent; 0.05% nonafluorovaleric acid, 흐름속도; 0.9 mL min−1)하에서 표준물 Se(IV), Se(VI), SeCys (selenocystein), SeMet (selenomethionine), Se-M-C (seleno methyl cystein)를 성공적으로 분리하였다. 시료에서의 셀레늄화학종의 추출은 마이크로파를 이용한 추출과 효소(protease I, trypsin,protease XIV)를 이용한 추출을 사용하였는데 시료에 따라 서로 다른 효율과 결과를 보여주었다. 식물성 시료인 마늘 시료는 protease I, 동물성 시료인 돼지고기와 고등어 시료는 protease I + trypsin이 가장 효율적인 추출을 보여주었다. 각 시료의 최적 조건을 선택하여 셀레늄 화학종을 추출하고 분리 정량한 결과 이들에는 주로 무기 Se, SeCys, SeMet이 수 μg g−1 내지 수 십 μg g−1 수준으로 포함되어 있음을 알 수 있었다.
The purpose of this research is to separate and quantitate selenium species in some food samples with HPLC-ICP-MS. Cation exchange chromatography showed efficient separation only for inorganic Se species while reversed phase ion pair chromatography showed good separation for both inorganic and organic Se species. C8 column (SymmetryshieldTM RP8, 3.5 μm, 4.6 × 150 mm) was used with optimum condition of 5% methanol mobile phase, 0.05% of nonafluorovaleric acid ion pairing reagent. Five standard Se species of Se(IV), Se(VI),SeCys(selenocystein), SeMet(selenomethionine) and Se-M-C(seleno methyl cystein) were separated successfully under the optimum condition (mobile phase; 5% methanol, ion-pairing reagent; 0.05% nonafluorovaleric acid,flow rate; 0.9 mL min−1). To extract Se species, microwave assisted and enzyme-assisted extraction methods were studied. In enzyme-assisted extraction method, protease I for garlic, protease I plus trypsin for pork and mackerel, and protease XIV for tuna showed the best extraction efficiency. With the optimum condition for each sample, it was found that mostly inorganic Se, SeCys and SeMet are present in the sample studied ranging from few μg g−1 to few tens of μg g−1.
O. Wada, N. Kurihara and N. Yamazaki, Jpn. J. Nutr. Assess., 10, 199-210 (1993).
J. L. Shisler, T. G. Senkevich, M. J. Berry and B. Moss, Science, 279, 102-105 (1998).
J. Virtamo, E. Valkeila, G. Algrhan, S. Punsar, K. K. Huttnen and M. J. Karvonen, Cancer, 60, 145-148 (1987).
K. Kose, P. Dogan and Y. Kardas, Biol. Trace Elem. Res., 53, 51-56 (1996).
O. A. Levander. Annu. Rev. Nutriti., 7, 227-250 (1987).
J. Far, H. Preud'homme and R. Lobinski, Anal. Chim. Acta, 657, 175-190 (2010).
M. Klein, L. Ouerdane, M. Bueno and F. Pannier, Metallomics, 3, 513-520 (2011).
J. M. Navarrete, L. C. Longoria, M. T. Martinez and L. Cabrera, J. Radioanal. Nucl. Chem., 271, 599-601 (2007).
P. C. Uden, Anal. Bioanal. Chem., 373, 422-431 (2002).
X. Dauchy, M. Potin-Gautier, A. Astruc and M. Astruc, Fresen. J. Anal. Chem., 348, 792 (1994).
H. Cho and Y. Pak, J. Kor. Chem. Soc., 55(3), 472-477 (2011).
T. D. Grant, M. Montes-Bayon, D. Leduc, M. W. Fricke, N. Terry and J. A. Caruso, J. Chromatogr. A, 1026, 159-166 (2004).
S. Kokarnig, D. Kuehnelt, M. Stiboller, U. Hartleb and K. A. Francesconi, Anal. Bioanal. Chem., 400, 2323- 2327 (2011).
Q. Chan, S. E. Afton and J. A. Caruso, J. Anal. Atom. Spectrom., 25, 186-192 (2010).
Q. Chan, S. E. Afton and J. A. Caruso, Metallomics, 2, 147-153 (2010).
C. B. Hymer and J. A. Caruso, J. Chromatogr. A, 1045, 1-13 (2004).
A. Prange and D. Pofrock, Anal. Bioanal. Chem., 383, 372-389 (2005).
T. DeSnaele, P. Verrept, L. Moens and R. Dams, Spectrochim. Acta. B, 50, 1409-1421 (1995).
L. H. Reyes, F. M. Sanz, P. H. Espilez, J. M. Marchante- Gayon, J. I. G. Alonso and A. Sanz-Medel, J. Anal. Atom. Spectrom., 19, 1230-1238 (2004).
Z. Pedrero, D. Elvira, C. Camara and Y. Madrid, Anal. Chim. Acta., 596, 251-256 (2007).
M. Kotrebai, M. Birringer, J. F. Tyson, E. Block and P. C. Uden, Analyst, 125, 71-78 (2000).
M. Shah, S. S. Kannamkunarath, J. C. A. Wuilloud, R. G. Wuilloud and J. A. Caruso, J. Anal. Atom. Spectrom., 19, 381-387 (2004).
C. Ip, M. Birringer, E. Block and M. Kotrebai, J. F. Tyson, P. C. Uden and D. J. Lisk, Agric. Food Chem., 48, 2062-2070 (2000).
C. Casiot, J. Szpunar, R. Lobinski and M. Potin-Gautier, J. Anal. Atom. Spectrom., 14, 645-652 (1999).
J. K. Kirby, G. H. Lyons and M. P. Karkkainen, J. Agric. Food Chem., 56(5), 1772-1779 (2008).
S. G. Casal, J. Far, K. Bierla, L. Ouerdane and J. Szpunar, Metallomics, 2(8), 535-548 (2010).
G. A. Zachariadis, D. C. Kapsimali and E. Rosenberg, Curr. Org. Chem., 14(19), 2282-2299 (2010).
S. Cho, M. Kim, Y. Kim, Y. Min and Y. Kim, Anal. Science & Tech., 19(3), 203-210 (2006).
O. Lee, Y. Jung and J. Moon, Korean J. Nutr., 43, 114- 122 (2010).
Y. Ham, B. Kim, Y. Kwon, H. Paik and S. Lee, Anal. Science & Tech., 24(3), 159-167 (2011).
S. Yoshida, M. Haratake, T. Fuchigami and M. Nakayama, J. Health Sci., 57, 215-224 (2011).
J. L. Guzma Mar, L. H. Reyes, G. M. M. Rahman and H. M. S. Kingston, J. Agric. Food Chem., 57, 3005- 3013 (2009).