Analysis of Estrogen in Pomegranate Extract by Solid Phase Extraction and Liquid Chromatography Tandem Mass Spectrometry

LC/MS/MS를 이용한 석류추출물 중의 에스트로겐 분석

  • 금은주 (한일양행 중앙연구소) ;
  • 권도형 (한일양행 중앙연구소) ;
  • 신혜승 (국립한경대학교 한경분석센터)
  • Received : 2010.03.09
  • Accepted : 2010.03.16
  • Published : 2010.03.31

Abstract

The pomegranate (Punica granatum), especially its fruit, possesses a vast ethnomedical history and represents a phytochemical reservoir of heuristic medical value. The tree and fruit can be divided into several anatomical compartments, and the fruit juice, peel and oil are known to be weakly estrogenic and heuristically of interest for treatment of menopausal symptoms and sequellae. In this study, analysis of estrogen in pomegranate extract was carried out with LC/MS/MS. Three batches of pomegranate extract samples were used to analysis the target compounds (estrogen). The contents of estrogen derivatives in the samples were 38.6 ppb of estriol, 83.5 ppb of estrone, and 10.9 ppb of estradiol. This result suggests that the pomegranate extract can used for treatment of menopause symptoms in the woman.

석류는 오래전부터 임상치료 및 식품원료로 사용되었으며 많은 피토에스트로겐 성분을 함유하고 있다. 그 성분으로는 에스트론, 에스트라디올, 에스트리올, 베타-시토스테롤, 엘라그산 등이 였으며 이러한 성분으로 인해 과즙, 껍질, 오일 등은 약한 에스트로겐 활성을 나타낸다고 보고되었으며, 이로 인해 석류는 여성 갱년기 치료용 원료로 많이 사용되고 있다. 그러나 에스트로겐 성분의 존재 유무에 대한 견해의 차이가 나타나고 있어 본 연구는 LC/MS/MS를 이용하여 석류추출물 중의 에스트로겐 분석을 통해 이를 확인하고자 하였으며, 간편하고 정확한 분석법을 모색하고자 하였다. 3개 롯트 시료의 에스트로겐을 분석한 결과, 에스트론 83.5ppb, 에스트라디올 10.9ppb 및 에스트리올 38.6ppb로 검출되었다.

Keywords

References

  1. Choi, D.W., Kim, J.Y., Choi, S.H., Jung, H.S., Kim, H.J., Cho, S.Y., Kang, C. S. and Chang, S.Y.: Identification of steroid hormones in pomegranate (Punica granatum) using HPLC and GC-mass spectromerty. Food Chem. 96, 562-571 (2006).
  2. Gil, M.I., Tomas-Barberan, F.A., Hess-Piearce, B., Holcroft D.M. and Kedar A.A.: Antioxidant activity of pomegranate juice and its relationship with phenolic composition and processing. J. Agric. Food Chem. 48, 4581-4589 (2000). https://doi.org/10.1021/jf000404a
  3. Seeram, N.P., Adams L.S., Henning, S.M., Niu, Y., Zhang, Y., Nair, M.G. and Heber D.: In vitro antiproliferative, apoptotic and antioxidant activities of punicalagin, ellagic acid and a total pomegranate tannin extract are enhanced in combination of other polyphenols. J. Nutr. Biochem. 16, 360-367 (2005). https://doi.org/10.1016/j.jnutbio.2005.01.006
  4. Afaq, F., Saleem, M., Krueger, C.G., Reed J.D. and Mukhtar, H.: Anthocyanins and hydrolyzable tannins rich pomegranate fruit extract modulates MAPKs and NF-kB pathways and inhibits skin tumorigenesis in CD-1 mice. Int. J. Cancer 113, 423-433 (2004).
  5. Lansky, E.P. and Newman R.A.: Punica granatum (pomegranate) and its potential for prevention and treatment of inflammation and cancer. J. ethnophar. 109, 177-206 (2007). https://doi.org/10.1016/j.jep.2006.09.006
  6. Choi, O.K., Chung K.S., Cho, G.S. Hwang, M.O. and Yoo, Y.S.: Proximate compositions and selected phytoestrogens of Iranian black pomegranate extract and its products. Korean J. Food and Nutr. 15(2), 119-125 (2002).
  7. Okamoto J.M., Hamamoto, Y.O., Yamoto, H. and Yoshimura, H.: Pomegranate extract improves a depressive state and bone properties in menopausal syndrome model ovariectomized mice. J. Ethnopharmacol. 92, 93-101 (2004). https://doi.org/10.1016/j.jep.2004.02.006
  8. Lee, Y.H., Hyun, S.H. and Choung, S.Y.: Effects of singled and mixed pomegranate on postmenopausal symptoms in ovariectomized rat. Yakhak Hoeji 50(3), 177-183 (2006).
  9. Adlercreutz, H., Markkanen H. and Watanabe, S.: Plasma concentrations of phyto-oestrogens in Japanese men. Lancet 342, 1209-1210 (1993). https://doi.org/10.1016/0140-6736(93)92188-Y
  10. Adlercreutz, H., Fotsis, T., Lampe, J., Wahala, K., Makela, T., Brunow, G. and Hase, T.: Quantitative determination of lignans and isoflavonoids in Plasma of omnivorous and vegetarian women by Isotope dilution gas ghromatography-mass spectrometry. J. Clin. Lab. Invest. 53, 5 (1993). https://doi.org/10.3109/00365519309090693
  11. Kim, S.H., Kim, I.H., Kang, B.H., Cha, T.Y., Lee, J.H., Rim, S.O., Song, K.S., Song, B.H., Kim, J.G. and Lee, J.M.: Analysis of extraction characteristics of phytoestrogen components from Punica granatum L. J. Korean Soc. Appl. Biol. Chem. 48(4), 352-357 (2005).
  12. Arao Y, Kanamori N, Kikkawa E, Otsuka H, Arimoto Y, Ikeda K, Inakuma T. and Kayama F.: A two-step screening method, using estrogen receptor-mediated trans activation, to measure estrogenicity in edible plants. Food Chem. 104, 1288- 1294 (2007). https://doi.org/10.1016/j.foodchem.2007.01.076
  13. Choi, S.Y., Lim, S.H., Kim, J.S., Ha, T.Y., Kim, S.R., Kang, K.S. and Hwang, I.K.: Evaluation of the estrogenic and antioxidant activity of some edible and medicinal plants. Korean J. food. Sci. Technol. 37(4), 549-556 (2005).
  14. Sturgeon S.R., Ronnenberg A.G.: Pomegranate and breast cancer: possible mechanisms of prevention. Nutrition Reviews 68(2), 122-128 (2009).
  15. Seeram N.P., Schulman R.N., Heber D, eds. Pomegranate- Ancient Roots of Modern Medicine. Boca Roton, FL: Taylor & Francis (2006).