Journal of Dairy Science and Biotechnology

Korean Society of Dairy Science and Biotechnology (한국낙농식품응용생물학회)
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In Vivo Evaluation of the Safety of Hydrolyzed GMP Powder containing Highly Concentrated Sialic Acid (23%) produced by Enzyme Separation and Solvent Enrichment Method using Micronucleus Test in Mice

효소분리 및 용매정제법으로 제조한 고농도 Sialic Acid(23%)가 함유된 GMP 가수분해분말의 마우스 골수세포의 소핵시험을 이용한 안전성 평가연구

  • Received : 2016.06.08
  • Accepted : 2016.06.20
  • Published : 2016.06.30
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Abstract

This study was designed to determine the mutagenic potential of hydrolyzed glycomacropeptide (GMP) powder (hereafter referred to as 23%-GNANA; product name: HELICOBACTROL-23) in a micronucleus test using bone marrow in ICR mice. Three experimental groups were used: a 3-step concentration group, with a maximum concentration of 2,000 mg/kg, and other sequentially two-fold lower concentrations, a negative control group, and a positive control group. The test material was administered for 2 d to observe the frequency of micronucleus formation up to 48 h after the test material was absorbed by the body. When the polychromatic erythrocyte (PCE) content of erythrocytes was compared, no significant differences were noted between the negative control group and the test group (p<0.05). Similarly, when the average numbers of micronucleated PCE (MNPCE) in 2,000 PCE per animal were compared, no significant difference was observed between the negative control group and the test group (p<0.05). No dose-response relationship with regard to the concentration of the test material administered was noted. These results allow us to conclude that hydrolyzed whey protein powder does not cause formation of micronuclei in mouse bone marrow cells under the applied conditions. In this study, the average frequency of micronucleus formation in PCE was significantly higher in the positive control group compared with the negative control group; thus, the test conditions were appropriate for detecting the frequency of micronucleus formation induced by the test material. In conclusion, the safety of 23%-GNANA test substance was verified in an in vivo micronucleus test in mice, conducted before the registration of HELICOBACTROL-23 as a food additive.

본 시험물질인 GMP으로부터 sialic acid를 분리 및 이의 농도를 23%로 높게 정제한 GMP 가수분해물(23%-GNANA) 분말에 대해 발암성 유발 유 무 판단의 기초자료를 얻기 위하여 마우스 골수세포를 이용한 소핵시험을 실시하였다. 결과로서, 다염성 적혈구의 소핵출현 적혈구의 출현비율은 음성 대조군과 시험물질 투여군간 평균 차이의 유의성이 없었다(p<0.05). 또한 동물개체별로 관찰한 결과에서도 음성대조군과 시험물질 투여군간 유의성이 없었으며(p<0.05), 시험물질의 용량의존성 반응은 전체적으로 나타나지 않았다. 이러한 결과로 보아, GMP로부터 제조한 23%-GNANA는 고농도로 sialic acid가 농축되었을 뿐, 일반 식품인 GMP 수준에서 마우스 골수세포에 대해 소핵의 유발성이 없는 것으로 판단되었다.

Keywords

References

  1. Ernest, P. B. 2000. Biological activities of bovine glycomacropeptide. British Journal of Nutrition 84:39-46.
  2. Fenench, M. and Morley, A. A. 1985. Measurement of micronuclei in lymphocyte. Mutation Research 147:29-36. https://doi.org/10.1016/0165-1161(85)90015-9
  3. Gorog, P. and Kovacs, I. B. 1978. Anti-inflammatory effect of sialic acid. Agents and Actions 8:543-545. https://doi.org/10.1007/BF02111443
  4. Heddle, J. A. A. 1973. Rapid in vivo test for chromosome damage. Mutation Research 18:187-190. https://doi.org/10.1016/0027-5107(73)90035-3
  5. Iijima, R., Takahashi, H., Namme, R., Ikegami, S. and Yamazaki, M. 2004. Novel biological funtion of sialic acid (N-acetyl-neuraminic acid) as a hydrogen peroxide scavenger. FEBS Letters 561:163-166. https://doi.org/10.1016/S0014-5793(04)00164-4
  6. Joskic, G. Nikolic, M. and Tisma, V. S. 1997. Radiosensitivity of different aged human lymphocytes following electron irradiation in vitro. Neoplasma 44(2):117-121.
  7. Ledeur, M. V. and Schmid. W. 1973.The micronucleus test methodological aspects. Mutation Research 19:109-117. https://doi.org/10.1016/0027-5107(73)90118-8
  8. Moon, Y. I., Lee, W. J. and Oh, S. J. 2005. Glycomacropeptide hydrolysed from bovine ${\kappa}$-casein; II. Chromatographic changes of ${\kappa}$-casein macropeptide as related to trichloroacetic acid concentration. Korean J. Food Sci. Ani. Resour. 17(1): 478-482.
  9. Schauer, R. and Wien, S. V. 1984. Sialic acids: Chemistry, metabolism and function. Carbohydrate Research 129:5-7. https://doi.org/10.1016/0008-6215(84)85324-0
  10. Schmid, W. 1975. The micronucleus test. Mutation Res. 31:9-15. https://doi.org/10.1016/0165-1161(75)90058-8
  11. Sudharsan, A. and Heddle, J. A. 1982. Simultaneous detection of chromosomal aberration and sister-chromatid exchanges. Toxicology and Applied Pharmacology 62:445-454. https://doi.org/10.1016/0041-008X(82)90145-4
  12. Wandl, E. O., Kain, R., Herbsthofer, T., Hienert, G. and Hobarth, G. 1989. Linear correlation between surviving fraction and the micronucleus frequency. J. Int. Radiat. Biol. 56(5): 771-775. https://doi.org/10.1080/09553008914552031
  13. Wang, B. 2009. Sialic acid is and essential nutrient for brain development and cognition. Annu. Rev. Nutr. 29: 177-222. https://doi.org/10.1146/annurev.nutr.28.061807.155515
  14. Wang, B., Brand-Miller, J., McVeagh, P. and Petocz, P. 2001. Concentration and distribution of sialic acid in human milk and infant formulas1-3. Am. J. Clin. Nutr. 74:510-515. https://doi.org/10.1093/ajcn/74.4.510
  15. Wang, B. and Brand-Miller. 2003. The role and potential of sialic acid in human nutrition. European Journal of Clinical Nutrition 57:1351-1369. https://doi.org/10.1038/sj.ejcn.1601704
  16. Wang, B., Yu, B., Karim, M., Hu, H., S., McGreevy, Y., Petocz, H. P., Held, S. and Miller, J. B. 2007. Dietary sialic acid supplementation improves learning and memory in piglets1-3. Am. J. Clin. Nutr. 85:561-569. https://doi.org/10.1093/ajcn/85.2.561
  17. Yoon, Y. C., Cho, J. K., Song, C. H., Lee, S. and Chung, C. I. 2000. Purification of the glycomacropeptide from cheese whey. Korean J. Food Sci. Ani. Resour. 20:159-165.