Research Article
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Comparative Determination of Melittin by Capillary Electrophoretic Methods

Year 2021, Volume: 8 Issue: 4, 1211 - 1216, 30.11.2021
https://doi.org/10.18596/jotcsa.949188

Abstract

Bee venom from honey bees (Apis Mellifera L.) is known to have many pharmacological and biological properties. Melittin, a peptide consisting of 26 amino acids, is known as the main component of bee venom. The study aims to develop a rapid capillary electrophoresis method for separating and quantifying melittin in honeybee venom. Since melittin is a basic peptide, it will adhere to the capillary wall during separation. Two different methods were developed in this study for the capillary electrophoretic separation of melittin. As a first approach, a low pH buffer system was used. For the second approach, the capillary column was coated with a positively charged polymer (PEI). With both methods developed, the migration of melittin in the capillary was achieved by preventing wall adsorption. Melittin migrated in 6 min when the low-pH buffer system was applied, whereas its migration time is longer than 10 min in the PEI-coated capillary column. Thus, a low-pH buffer system was preferred for the analysis of the actual bee-venom sample. 100 mmol L-1 phosphoric acid/sodium dihydrogen phosphate system at pH 1.55 was chosen as separation buffer. As a conclusion, a fast and reliable method was developed for the determination of melittin in honeybee venom. The method was applied to an Anatolian bee venom sample to highlight the melittin amount. The melittin amount was found as 24.5 ± 3.4 g 100 g-1 in the bee venom sample.

Supporting Institution

İstanbul Teknik Üniversitesi Bilimsel Araştırma Projeleri Birimi

Thanks

The authors would like to thank Assoc. Prof. Dr. Sibel Döşler for giving away the standard melittin.

References

  • 1. Gupta RK, Stangaciu S. Apitherapy: Holistic Healing Through the Honeybee and Bee Products in Countries with Poor Healthcare System. In: Gupta RK, Reybroeck W, van Veen JW, Gupta A, editors. Beekeeping for Poverty Alleviation and Livelihood Security [Internet]. Dordrecht: Springer Netherlands; 2014 [cited 2021 Nov 6]. p. 413–46. ISBN: 978-94-017-9199-1.
  • 2. Luo X, Dong Y, Gu C, Zhang X, Ma H. Processing Technologies for Bee Products: An Overview of Recent Developments and Perspectives. Front Nutr. 2021 Nov 3;8:727181.
  • 3. Kalaycıoğlu Z, Kaygusuz H, Döker S, Kolaylı S, Erim FB. Characterization of Turkish honeybee pollens by principal component analysis based on their individual organic acids, sugars, minerals, and antioxidant activities. LWT. 2017 Oct;84:402–8.
  • 4. Kaygusuz H, Tezcan F, Erim FB, Yildiz O, Sahin H, Can Z, et al. Characterization of Anatolian honeys based on minerals, bioactive components and principal component analysis. LWT-Food Science and Technology. 2016 May;68:273–9.
  • 5. Kaygusuz H. Analysis of a Rare Honey Sample From Tuzluca/Iğdır Region. Journal of the Institute of Science and Technology. 2020 Jun 1;1139–45.
  • 6. Sarikaya AO, Ulusoy E, Öztürk N, Tunçel M, Kolayli S. Antioxidant Activity And Phenolic Acid Constituents Of Chestnut (Castania Sativa Mill.) Honey And Propolis. Journal of Food Biochemistry. 2009 Aug;33(4):470–81.
  • 7. Ali M. Studies on Bee Venom and Its Medical Uses. 2012;1:1–15.
  • 8. Bellik Y. Bee Venom: Its Potential Use in Alternative Medicine. AIA. 2015 May 20;13(1):3–16.
  • 9. Wehbe R, Frangieh J, Rima M, El Obeid D, Sabatier J-M, Fajloun Z. Bee Venom: Overview of Main Compounds and Bioactivities for Therapeutic Interests. Molecules. 2019 Aug 19;24(16):2997.
  • 10. Uzuner SÇ, Birinci E, Tetikoğlu S, Birinci C, Kolaylı S. Distinct Epigenetic Reprogramming, Mitochondrial Patterns, Cellular Morphology, and Cytotoxicity after Bee Venom Treatment. PRA. 2021 Aug;16(3):377–92.
  • 11. Silva J, Monge-Fuentes V, Gomes F, Lopes K, Anjos L, Campos G, et al. Pharmacological Alternatives for the Treatment of Neurodegenerative Disorders: Wasp and Bee Venoms and Their Components as New Neuroactive Tools. Toxins. 2015 Aug 18;7(8):3179–209.
  • 12. Kim W. Bee Venom and Its Sub-Components: Characterization, Pharmacology, and Therapeutics. Toxins. 2021 Mar 7;13(3):191.
  • 13. Alia O, Laila M, Antonious A. Antimicrobial Effect of Melittin Isolated From Syrian Honeybee (Apis Mellifera) Venom and Its Wound Healing Potential. Int J Pharm Sci Rev Res. 2013;21:318–24.
  • 14. Raghuraman H, Chattopadhyay A. Melittin: a Membrane-active Peptide with Diverse Functions. Bioscience Reports. 2007 Aug 6;27(4–5):189–223.
  • 15. Rady I, Siddiqui IA, Rady M, Mukhtar H. Melittin, a major peptide component of bee venom, and its conjugates in cancer therapy. Cancer Letters. 2017 Aug;402:16–31.
  • 16. Haghi G, Hatami A, Mehran M. Qualitative and quantitative evaluation of melittin in honeybee venom and drug products containing honeybee venom. Journal of Apicultural Science. 2013 Dec 1;57(2):37–44.
  • 17. Huang S, Wang J, Guo Z, Wang Y, Liu C. Quantitative Measurement of Melittin in Asian Honeybee Venom Using a New Method Including UPLC-QqTOF-MS. Toxins. 2020 Jul 4;12(7):437.
  • 18. Ionete R, Dinca O, Tamaian R, Geana E. Exploring Apis Mellifera Venom Compounds Using Highly Efficient Methods. Progress of Cryogenics and Isotopes Separation. 2013;16:89–100.
  • 19. Kokot ZJ, Matysiak J. Simultaneous Determination of Major Constituents of Honeybee Venom by LC-DAD. Chroma. 2009 Jun;69(11–12):1401–5.
  • 20. Rybak-Chmielewska H, Szczêsna T. HPLC Study of Chemical Composition of Honeybee (Apis Mellifera L.) Venom. J Apic Sci. 2004;48:103–9.
  • 21. Samanci T, Kekeçoğlu M. Comparison of Commercial and Anatolian Bee Venom in Terms of Chemical Composition. Uludağ Arıcılık Dergisi. 2019 May 29;61–8.
  • 22. Kokot ZJ, Matysiak J, Urbaniak B, Dereziński P. New CZE-DAD method for honeybee venom analysis and standardization of the product. Anal Bioanal Chem. 2011 Mar;399(7):2487–94.
  • 23. Barwick V, editor. Eurachem/CITAC Guide: Guide to Quality in Analytical Chemistry: An Aid to Accreditation [Internet]. Available from: www.eurachem.org. ISBN: 978-0-948926-32-7.
  • 24. Chen F-TA, Evangelista RA. Protein analysis by capillary electrophoresis. In: Shintani H, Polonský J, editors. Handbook of Capillary Electrophoresis Applications [Internet]. Dordrecht: Springer Netherlands; 1997 [cited 2021 Nov 6]. p. 173–97. ISBN: 978-94-009-1561-9.
  • 25. Erim FB, Cifuentes A, Poppe H, Kraak JC. Performance of a physically adsorbed high-molecular-mass polyethyleneimine layer as coating for the separation of basic proteins and peptides by capillary electrophoresis. Journal of Chromatography A. 1995 Aug;708(2):356–61.
Year 2021, Volume: 8 Issue: 4, 1211 - 1216, 30.11.2021
https://doi.org/10.18596/jotcsa.949188

Abstract

References

  • 1. Gupta RK, Stangaciu S. Apitherapy: Holistic Healing Through the Honeybee and Bee Products in Countries with Poor Healthcare System. In: Gupta RK, Reybroeck W, van Veen JW, Gupta A, editors. Beekeeping for Poverty Alleviation and Livelihood Security [Internet]. Dordrecht: Springer Netherlands; 2014 [cited 2021 Nov 6]. p. 413–46. ISBN: 978-94-017-9199-1.
  • 2. Luo X, Dong Y, Gu C, Zhang X, Ma H. Processing Technologies for Bee Products: An Overview of Recent Developments and Perspectives. Front Nutr. 2021 Nov 3;8:727181.
  • 3. Kalaycıoğlu Z, Kaygusuz H, Döker S, Kolaylı S, Erim FB. Characterization of Turkish honeybee pollens by principal component analysis based on their individual organic acids, sugars, minerals, and antioxidant activities. LWT. 2017 Oct;84:402–8.
  • 4. Kaygusuz H, Tezcan F, Erim FB, Yildiz O, Sahin H, Can Z, et al. Characterization of Anatolian honeys based on minerals, bioactive components and principal component analysis. LWT-Food Science and Technology. 2016 May;68:273–9.
  • 5. Kaygusuz H. Analysis of a Rare Honey Sample From Tuzluca/Iğdır Region. Journal of the Institute of Science and Technology. 2020 Jun 1;1139–45.
  • 6. Sarikaya AO, Ulusoy E, Öztürk N, Tunçel M, Kolayli S. Antioxidant Activity And Phenolic Acid Constituents Of Chestnut (Castania Sativa Mill.) Honey And Propolis. Journal of Food Biochemistry. 2009 Aug;33(4):470–81.
  • 7. Ali M. Studies on Bee Venom and Its Medical Uses. 2012;1:1–15.
  • 8. Bellik Y. Bee Venom: Its Potential Use in Alternative Medicine. AIA. 2015 May 20;13(1):3–16.
  • 9. Wehbe R, Frangieh J, Rima M, El Obeid D, Sabatier J-M, Fajloun Z. Bee Venom: Overview of Main Compounds and Bioactivities for Therapeutic Interests. Molecules. 2019 Aug 19;24(16):2997.
  • 10. Uzuner SÇ, Birinci E, Tetikoğlu S, Birinci C, Kolaylı S. Distinct Epigenetic Reprogramming, Mitochondrial Patterns, Cellular Morphology, and Cytotoxicity after Bee Venom Treatment. PRA. 2021 Aug;16(3):377–92.
  • 11. Silva J, Monge-Fuentes V, Gomes F, Lopes K, Anjos L, Campos G, et al. Pharmacological Alternatives for the Treatment of Neurodegenerative Disorders: Wasp and Bee Venoms and Their Components as New Neuroactive Tools. Toxins. 2015 Aug 18;7(8):3179–209.
  • 12. Kim W. Bee Venom and Its Sub-Components: Characterization, Pharmacology, and Therapeutics. Toxins. 2021 Mar 7;13(3):191.
  • 13. Alia O, Laila M, Antonious A. Antimicrobial Effect of Melittin Isolated From Syrian Honeybee (Apis Mellifera) Venom and Its Wound Healing Potential. Int J Pharm Sci Rev Res. 2013;21:318–24.
  • 14. Raghuraman H, Chattopadhyay A. Melittin: a Membrane-active Peptide with Diverse Functions. Bioscience Reports. 2007 Aug 6;27(4–5):189–223.
  • 15. Rady I, Siddiqui IA, Rady M, Mukhtar H. Melittin, a major peptide component of bee venom, and its conjugates in cancer therapy. Cancer Letters. 2017 Aug;402:16–31.
  • 16. Haghi G, Hatami A, Mehran M. Qualitative and quantitative evaluation of melittin in honeybee venom and drug products containing honeybee venom. Journal of Apicultural Science. 2013 Dec 1;57(2):37–44.
  • 17. Huang S, Wang J, Guo Z, Wang Y, Liu C. Quantitative Measurement of Melittin in Asian Honeybee Venom Using a New Method Including UPLC-QqTOF-MS. Toxins. 2020 Jul 4;12(7):437.
  • 18. Ionete R, Dinca O, Tamaian R, Geana E. Exploring Apis Mellifera Venom Compounds Using Highly Efficient Methods. Progress of Cryogenics and Isotopes Separation. 2013;16:89–100.
  • 19. Kokot ZJ, Matysiak J. Simultaneous Determination of Major Constituents of Honeybee Venom by LC-DAD. Chroma. 2009 Jun;69(11–12):1401–5.
  • 20. Rybak-Chmielewska H, Szczêsna T. HPLC Study of Chemical Composition of Honeybee (Apis Mellifera L.) Venom. J Apic Sci. 2004;48:103–9.
  • 21. Samanci T, Kekeçoğlu M. Comparison of Commercial and Anatolian Bee Venom in Terms of Chemical Composition. Uludağ Arıcılık Dergisi. 2019 May 29;61–8.
  • 22. Kokot ZJ, Matysiak J, Urbaniak B, Dereziński P. New CZE-DAD method for honeybee venom analysis and standardization of the product. Anal Bioanal Chem. 2011 Mar;399(7):2487–94.
  • 23. Barwick V, editor. Eurachem/CITAC Guide: Guide to Quality in Analytical Chemistry: An Aid to Accreditation [Internet]. Available from: www.eurachem.org. ISBN: 978-0-948926-32-7.
  • 24. Chen F-TA, Evangelista RA. Protein analysis by capillary electrophoresis. In: Shintani H, Polonský J, editors. Handbook of Capillary Electrophoresis Applications [Internet]. Dordrecht: Springer Netherlands; 1997 [cited 2021 Nov 6]. p. 173–97. ISBN: 978-94-009-1561-9.
  • 25. Erim FB, Cifuentes A, Poppe H, Kraak JC. Performance of a physically adsorbed high-molecular-mass polyethyleneimine layer as coating for the separation of basic proteins and peptides by capillary electrophoresis. Journal of Chromatography A. 1995 Aug;708(2):356–61.
There are 25 citations in total.

Details

Primary Language English
Subjects Analytical Chemistry
Journal Section Articles
Authors

Melda Akay 0000-0002-5272-8656

Zeynep Kalaycıoğlu 0000-0002-0967-0997

Sevgi Kolaylı 0000-0001-9969-4177

Bedia Berker 0000-0001-9406-6681

Publication Date November 30, 2021
Submission Date June 8, 2021
Acceptance Date November 5, 2021
Published in Issue Year 2021 Volume: 8 Issue: 4

Cite

Vancouver Akay M, Kalaycıoğlu Z, Kolaylı S, Berker B. Comparative Determination of Melittin by Capillary Electrophoretic Methods. JOTCSA. 2021;8(4):1211-6.