Effects of various lights, solvents, and zinc protoporphyrin on the chemical behavior of MTT formazan

빛, 용매와 zinc protoporphyrin에 의한 MTT 포마잔의 화학적 동태 변화

  • Kim, Joo Hyoun (Division of Applied Food System, College of Natural Science, Seoul Women's University) ;
  • Hong, Jungil (Division of Applied Food System, College of Natural Science, Seoul Women's University)
  • 김주현 (서울여자대학교 자연과학대학 식품응용시스템학부) ;
  • 홍정일 (서울여자대학교 자연과학대학 식품응용시스템학부)
  • Received : 2017.07.12
  • Accepted : 2017.09.13
  • Published : 2018.02.28


The MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay is commonly used for analyzing the cell viability. In this study, effects of various solvents, different lights, and zinc protoporphyrin (ZnPP) on the chemical behavior of MTT formazan were investigated. The color response of MTT formazan in NaOH was highly pronounced; the absorbance of MTT formazan in 0.1 N NaOH at 550 nm was >2-fold higher than that in water, dimethyl sulfoxide (DMSO), methanol, and ethanol. MTT formazan in DMSO and NaOH (>0.1 N) was relatively stable under fluorescent and UV light at 365 nm; its rapid degradation was induced under UV light at 254 nm in all solvents. ZnPP degraded MTT formazan under light in a time- and concentration-dependent manner; MTT formazan in 0.1 N NaOH was the most sensitive to ZnPP, followed by DMSO. These results suggest that NaOH and DMSO might be suitable media for MTT formazan for monitoring photosensitizing properties.


MTT formazan;zinc protoporphyrin;light;sodium hydroxide;photosensitizing property


Supported by : 한국연구재단


  1. Berridge MV, Herst PM, Tan AS. Tetrazolium dyes as tools in cell biology: New insights into their cellular reduction. Biotechnol. Annu. Rev. 11: 127-152 (2005)
  2. Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J. Immunol. Methods 65: 55-63 (1983)
  3. Gerlier D, Thomasset N. Use of MTT colorimetric assay to measure cell activation. J. Immunol. Methods 94: 57-63 (1986)
  4. Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB. Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 47: 936-942 (1987)
  5. Twentyman PR, Luscombe M. A study of some variables in a tetrazolium dye (MTT) based assay for cell growth and chemosensitivity. Brit. J. Cancer 56: 279-285 (1987)
  6. Plumb JA, Milroy R and Kaye S. Effects of the pH dependence of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromideformazan absorption on chemosensitivity determined by a novel tetrazolium-based assay. Cancer Res. 49: 4435-4440 (1989)
  7. Bruggisser R, von Daeniken K, Jundt G, Schaffner W, Tullberg-Reinert H. Interference of plant extracts, phytoestrogens and antioxidants with the MTT tetrazolium assay. Planta Med. 68: 445-448 (2002)
  8. Talorete TP, Bouaziz M, Sayadi S, Isoda H. Influence of medium type and serum on MTT reduction by flavonoids in the absence of cells. Cytotechnology 52: 189-198 (2006)
  9. Park KA, Choi HA, Kim MR, Choi YM, Kim HJ, Hong J. Changes in color responses of MTT formazan by zinc protoporphyrin. Korean J. Food Sci. Technol. 43: 754-759 (2011)
  10. Chakrabarti R, Kundu S, Kumar S, Chakrabarti R. Vitamin A as an enzyme that catalyzes the reduction of MTT to formazan by vitamin C. J. Cell. Biochem. 80: 133-138 (2000)
  11. Peng L, Wang B, Ren P. Reduction of MTT by flavonoids in the absence of cells. Colloid. Surface. B. 45: 108-111 (2005)
  12. Bonkovsky HL, Guo JT, Hou W, Li T, Narang T, Thapar M. Porphyrin and heme metabolism and the porphyrias. Compr. Physiol. 3: 365-401 (2013)
  13. Biesaga M, Pyrzynska K, Trojanowicz M. Porphyrins in analytical chemistry. A review. Talanta 51: 209-224 (2000)
  14. Labbe RF, Vreman HJ, Stevenson DK. Zinc protoporphyrin: A metabolite with a mission. Clin. Chem. 45: 2060-2072 (1999)
  15. Benedini R, Raja V, Parolari G, Zinc-protoporphyrin IX promoting activity in pork muscle. LWT-Food Sci. Technol. 41: 1160-1166 (2008)
  16. Lee T, Chau L. Heme Oxygenase-1 Mediates the Anti-Inflammatory Effect of Interleukin-10 in Mice. Nat. Med. 8: 240-246 (2002)
  17. Tanaka S, Akaike T, Fang J, Beppu T, Ogawa M, Tamura F, Miyamoto Y, Maeda H. Antiapoptotic effect of haem oxygenase-1 induced by nitric oxide in experimental solid tumour. Brit. J. Cancer 88: 902-909 (2003)
  18. Kennedy JC, Pottier RH. New trends in photobiology: Endogenous protoporphyrin IX, a clinically useful photosensitizer for photodynamic therapy. J. Photoch. Photobio. B 14: 275-292 (1992)
  19. Hansen MB, Nielsen, SE, Berg K. Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell killing. J. Immunol. Methods 119: 203-211 (1989)
  20. Abe K, Matsuki N. Measurement of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction activity and lactate dehydrogenase release using MTT. Neurosci. Res. 38:325-329 (2000)
  21. Edmondson JM, Armstrong LS, Martinez AO. A rapid and simple MTT-based spectrophotometric assay for determining drug sensitivity in monolayer cultures. J. Tissue Cult. Meth. 11: 15-17 (1988)
  22. Plumb JA. Cell sensitivity assays: The MTT assay. pp. 165-169. In: Cancer cell culture: methods and protocols. Langdon SP (ed). Humana Press Inc., Totowa, NJ, USA (2004)
  23. Choi BH. Modulation of color response in the tetrazolium-based assay by porphyrins and polyphenols, MS thesis, Seoul Women's University, Seoul, Korea (2015)
  24. Roehm NW, Rodgers GH, Hatfield SM, Glasebrook AL. An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT. J. Immunol. Methods 142: 257-265 (1991)