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Research Articles






M. I. Udonkang , Kommomo A Ubi, Imeobong J Inyang 



Formalin is a popular fixative and biohazard in histology and pathology laboratories. The objectives were to investigate the physiochemical properties and effect of different concentrations of honey as fixative on gross morphology and histology of tissues. Honey’s glucose concentration and pH were measured. Five organs from goat were gotten in pairs and small tissues cut before being grouped into A and B. Group A were fixed in 20%, 50%, 70%, 90%,100% honey concentrations while group B were fixed in 10% buffered formal saline for 48 hours-6 months. The cut tissues were processed by paraffin wax-embedding method and stained with haematoxylin and eosin. Glucose concentration was 284mmol/l and pH was 4.0. Macroscopically, tissues fixed in 20% and 50% honey showed putrefaction changes after 72 hours and were statistically significant (p=0.004). Tissues preserved in 70%, 90% and 100% honey were properly fixed for up to 6 months and beyond. Minor differences in nuclear and cytoplasmic staining (p=0.391), intensity and clarity of histological details (p=0.252) among the honey-fixed group were not statistically significant. In conclusion, the 70-100% honeys are suitable for long term gross preservation while 20-50% concentrations give excellent tissue staining characteristics. Thus, honey is a safer substitute for formalin.

KEY WORDS: Formalin, honey, fixation, histology, substitute


  1. Patil S, Rao R. S, Gonari S, Majumdar B. Natural sweetners as fixatives in histopathology: A longitudinal study. Journal of Natural Science, Biology and Medicine, 6(1):67-70.

  2. Thavarajah R, Mudimbaimannar V. K, Elizabeth J, Rao U. K, Ranganathan K. Chemical and physical basics of routine formaldehyde fixation. Journal of Oral and Maxillofacial Pathology, 2012; 16: 400-405.

  3. Wang S, Cui X, Fang G. Rapid determination of formaldehyde and sulfur dioxide in food products and Chinese herbals. Food Chemistry, 2007; 103: 1487-1493.

  4. Agency for Toxic Substances and Disease Registry (ATSDR). Medical management for formaldehyde (HCHO). Centre for Disease Control, Atlanta, 2014: 1-24.

  5. Onyije F. M, Avwioro O. G, Excruciating effects of formaldehyde exposure to students in gross anatomy dissection laboratory. International Journal of Occupational and Environmental Medicine, 2012; 3: 92-95.

  6. Brenner E. Human body preservation-old and new techniques. Journal of Anatomy, 2014; 224: 316-344.

  7. Al-Maaini R, Bryant P. The effectiveness of honey as a substitute for formalin in histological fixation of tissue. Journal of Histotechniques, 2006; 29: 173-176.

  8. WHO. World Health Organisation, guidelines for indoor air quality: selected pollutants. Formaldehyde. 2010: 103-144. index.html

  9. Al-Maaini R, Bryant P. Honey as an alternative to formalin in the demonstration of connective tissue components. Journal of Histotechniques, 2008; 31: 67-72.

  10. Ishaq R, Azmat H, Omair M, Shiekh A. K, Tanwani A. K. Comparison of honey with alcohol as a fixative in fine needle aspiration cytology. International Journal of Pathology, 2016; 14: 1-4.

  11. Nwaeze J. A, Okafor J. I, Nwaeze E. I, Nwaeze J. E. Evaluation of physicochemical and antioxidant properties of two stingless bee honeys: a comparison with Apis mellifera honey from Nsukka, Nigeria. BMC Research Notes, 2017; 10: 566.

  12. Majumdar B, Rao R. S, Patil S. Tissue preservation with natural fixatives: An immunohistochemical evaluation. World Journal of Dentistry, 2016; 7: 87-91.

  13. Igwe EC, Dandago MA, Binga EN. Assessment of quality attributes of natural honey from Adamawa state North Eastern, Nigeria. African Journal of Food Sciences, 2012; 6: 449-455.

  14. Bancroft J. D, Gamble M. Theory and Practice of Histological Techniques. 6th ed. Edinburgh: Churchill Livingstone, 2008: 56-59.

  15. Geiling N. The science behind honey’s eternal shelf life,, August, 22, 2013. Retrieved online on 26 August, 2018.

  16. Saores S, Amaral J. S, Oliveira MB, Matra I. A comprehensive review on the main honey authentication issues: Production and origin. Compreh. Rev. Food Sci. Food Safety, 2017; 16: 1072-1100.

  17. James O. O, Mesubi M. A, Usman L. A, Yeye S. O, Ajanaku K. O, Ogunniran K. O, et al. Physical characterization of some honey samples from North-Central Nigeria. International Journal of Physical Sciences, 2009; 4: 464-470.

  18. Bhattacharyya A, Gupta B, Singh A, Sah K, Gupta V. Probing natural substitute for formalin: Comparing honey, sugar, and jiggery syrup as fixatives. National Journal of Maxillofacial Surgery, 2018; 9: 14-21.

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