5-Hydroxymethyl Furfural (HMF) Evaluation in Molasses Produced through Different Traditional Methods Using HPLC

Pınar Karagül; Elif Sine Düvenci; Bora Karagül

doi:10.18596/jotcsa.1799306

5-Hydroxymethyl Furfural (HMF) Evaluation in Molasses Produced through Different Traditional Methods Using HPLC

Abstract

Molasses is a widely consumed traditional food product obtained from various fruits and valued for its nutritional and sensory properties. Thermal processing during molasses production promotes Maillard reactions, which contribute to desirable color and flavor development but may also result in the formation of undesirable compounds such as 5-hydroxymethylfurfural (HMF). Elevated HMF levels are associated with high processing temperatures and prolonged storage periods and are considered indicators of food quality and safety. In this study, a validated high-performance liquid chromatography–photodiode array detection (HPLC–PDA) method was employed to determine HMF levels in grape, mulberry, and carob molasses produced by traditional methods. The method demonstrated excellent linearity (R² = 0.9993) within the calibration range of 6.25–50 µg/mL. The limits of detection (LOD) and quantification (LOQ) were determined as 5.93 µg/mL and 17.97 µg/mL, respectively. HMF was quantified at 26.24 µg/mL in carob molasses, while grape and mulberry molasses contained HMF levels below the LOQ. In addition, total phenolic content (TPC) and total flavonoid content (TFC) were evaluated. Carob molasses exhibited the highest TPC (748.59 mg/L) and TFC (270.64 mg/L), followed by mulberry and grape molasses. Statistical analysis revealed significant differences among molasses types (p<0.05). The findings indicate that traditional production practices may affect HMF formation. Regular monitoring of HMF and optimization of production conditions are important to ensure product quality and consumer safety.

Keywords

Thanks

The HPLC analyses in this study were carried out at Düzce University Scientific and Technological Research Application and Research Centre (DUBİT).

References

1. Feliatra F, Hendra M, Batubara UM, Effendi I, Adelina A, Feliatra VA. Potential of single cell protein production using waste as growth medium. IOP Conf Ser Earth Environ Sci [Internet]. 2022 Dec 1;1118(1):012024. Available from: .
2. Daanaa HSA, Abdou M, Goda HA, Abbas MT, Hamza MA, Sarhan MS, et al. Plant pellets: A compatible vegan feedstock for preparation of plant-based culture media and production of value-added biomass of rhizobia. Sustainability [Internet]. 2020 Oct 12;12(20):8389. Available from: .
3. Kitamura DH, Vandenberghe LP de S, Rodrigues C, Salmon DNX, Pereira GV de M, Soccol CR. Selenium-enriched probiotic Saccharomyces boulardii CCT 4308 biomass production using low-cost sugarcane molasses medium. Brazilian Arch Biol Technol [Internet]. 2021;64:e21200658. Available from: .
4. Behravan J, Fazly Bazzaz BS, Salimi Z. Optimization of dextran production by Leuconostoc mesenteroides NRRL B‐512 using cheap and local sources of carbohydrate and nitrogen. Biotechnol Appl Biochem [Internet]. 2003 Dec 23;38(3):267–9. Available from: .
5. Palmonari A, Federiconi A, Cavallini D, Sniffen CJ, Mammi L, Turroni S, et al. Impact of molasses on ruminal volatile fatty acid production and microbiota composition in vitro. Animals [Internet]. 2023 Feb 17;13(4):728. Available from: .
6. Tüzün S, Baş İ, Karakavuk E, Karaca Sanyürek N, Benzer F. Çeşitli pekmez türlerinde farklı yöntemlerle tespit edilen antioksidan aktivitelerin karşılaştırılması. Türk Tarım ve Doğa Bilim Derg [Internet]. 2020 Apr 24;7(2):323–30. Available from: .
7. Denk E. Erzurum’un karnavas dut pekmezi: Gastronomik miras ve coğrafi işaretle buluştuğu noktada gelenek ve yenilik arasındaki diyalektik. J Tour Gastron Stud [Internet]. 2023 Sep 30;11(3):2418–47. Available from: .
8. Kıran TR, Otlu O, Karabulut E, Bay Karabulut A. Protective effects of grape molasses and resveratrol against DMBA induced oxidative stress in rat ovarian tissues. Middle Black Sea J Heal Sci [Internet]. 2019 Aug 28;5(2):151–9. Available from: .

9. Akan LS. Production and characteristics of a traditional food: Molasses (Pekmez). Food Sci Nutr Stud [Internet]. 2018 Oct 4;2(2):25–32. Available from: .
10. Çetinkaya T, Karacaoğlan V, Bayıl Oguzkan S. Kastamonu yöresinde üretilen göknar kozalak pekmez ve sıvı şuruplarının bazı fiziksel ve kimyasal özelliklerinin karşılaştırılması. Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Derg [Internet]. 2024 Jun 30;27(3):685–94. Available from: .
11. Yildiz Kemer D, İşler Dalgiç A. Complementary health approaches for the symptoms of pediatric oncology patients. Holist Nurs Pract [Internet]. 2023 Mar;37(2):98–109. Available from: .
12. Aydın S, Özdemir Y. Development and characterization of carob flour based functional spread for increasing use as nutritious snack for children. J Food Qual [Internet]. 2017;2017:5028150. Available from: .
13. Toufeili I, Itani M, MonaZeidan M, Yamani O Al, Kharroubi S. Nutritional and functional potential of carob syrup versus date and maple syrup. Food Technol Biotechnol [Internet]. 2022 Jul 15;60(2):266–78. Available from: .
14. El-Sayed MI, Aly E, El-Deeb AM. Improving the physicochemical and antioxidative properties of fermented goat milk using carob molasses and some probiotic strains. Food Sci Biotechnol [Internet]. 2024 Feb 25;33(3):657–66. Available from: .
15. Lakkab I, Ouakil A, El Hajaji H, Lachkar N, Lefter R, Ciobica A, et al. Carob seed peels effect on cognitive impairment and oxidative stress status in methionine-induced mice models of schizophrenia. Brain Sci [Internet]. 2022 Dec 3;12(12):1660. Available from: .
16. Gadge AS, Shirsat D V., Soumia PS, Pote CL, Pushpalatha M, Pandit TR, et al. Physiochemical, biological, and therapeutic uses of stingless bee honey. Front Sustain Food Syst [Internet]. 2024 Jan 4;7:1324385. Available from: .
17. Shapla UM, Solayman M, Alam N, Khalil MI, Gan SH. 5-Hydroxymethylfurfural (HMF) levels in honey and other food products: effects on bees and human health. Chem Cent J [Internet]. 2018 Dec 4;12(1):35. Available from: .
18. Türköz Acar E, Helvacıoğlu S, Charehsaz M, Aydın A. Determination and safety evaluation of furfural and hydroxymethylfurfural in some honey samples by using a validated HPLC-DAD method. J Res Pharm [Internet]. 2018 Jul 2;22(4):519–27. Available from: .
19. Toker OS, Dogan M, Ersöz NB, Yilmaz MT. Optimization of the content of 5-hydroxymethylfurfural (HMF) formed in some molasses types: HPLC-DAD analysis to determine effect of different storage time and temperature levels. Ind Crops Prod [Internet]. 2013 Oct;50:137–44. Available from: .
20. Oral RA, Dogan M, Sarioglu K, Toker ÖS. 5-hydroxymethyl furfural formation and reaction kinetics of different pekmez samples: effect of temperature and storage. Int J Food Eng [Internet]. 2012 Sep 14;8(4). Available from: .
21. Şanli S, Altınbaş S. Determination of benzoic acid, sorbic acid and HMF in grape molasses produced by traditional and modern methods. J Pharm Res Int [Internet]. 2021 Jul 28;33(38B):330–6. Available from: .
22. Spano N, Casula L, Panzanelli A, Pilo M, Piu P, Scanu R, et al. An RP-HPLC determination of 5-hydroxymethylfurfural in honeyThe case of strawberry tree honey. Talanta [Internet]. 2006 Feb 15;68(4):1390–5. Available from: .
23. Elmastas A, Aydin F, Umaz A, Kılınc E, Arslan Y, Aydin I. Determination of hydroxymethylfurfural in Turkish honeys, pekmez (grape molasses), and jam samples by high‐performance liquid chromatography with diode array detection. J Food Process Preserv [Internet]. 2022 Apr 17;46(4):e16462. Available from: .
24. Rosatella AA, Simeonov SP, Frade RFM, Afonso CAM. 5-Hydroxymethylfurfural (HMF) as a building block platform: Biological properties, synthesis and synthetic applications. Green Chem [Internet]. 2011;13(4):754–93. Available from: .
25. Moghaddam HN, Alaeepajouh S, Behzad M, Hajimahmoodi M, Sadeghi N. Evaluating the antioxidant activity and the level of 5-hydroxymethylfurfural in honey. J Food Saf Hyg [Internet]. 2023 Feb 27;9(4):241–51. Available from: .
26. Osanmaz A. HMF analysis in apple juice by HPLC. Gıda Teknol Derg. 2021;42–5.
27. Slinkard K, Singleton VL. Total phenol analysis: Automation and comparison with manual methods. Am J Enol Vitic [Internet]. 1977;28(1):49–55. Available from: .
28. Chandra S, Khan S, Avula B, Lata H, Yang MH, ElSohly MA, et al. Assessment of total phenolic and flavonoid content, antioxidant properties, and yield of aeroponically and conventionally grown leafy vegetables and fruit crops: A comparative study. Ohta Y, editor. Evidence-Based Complement Altern Med [Internet]. 2014 Jan 23;2014(1):253875. Available from: .
29. Düvenci ES, Karagül P, Karagül B. Determination of caffeine contents in soft drinks by high performance liquid chromatography (HPLC) method. Akad Gıda [Internet]. 2025 Sep 30;23(3):203–9. Available from: .
30. Horwitz W. AOAC guidelines for single laboratory validation of chemical methods for dietary supplements and botanicals. AOAC Int Gaithersburg, MD, USA. 2002;1219.
31. Özesmer B, Özsisli B. Geleneksel ve endüstriyel yöntemlerle üretilen bazı üzüm pekmezlerinin fizikokimyasal ve duyusal özelliklerinin karşılaştırılması. Turkish J Agric - Food Sci Technol [Internet]. 2025 Sep 27;13(9):2749–58. Available from: .
32. Altiok D, Şen L, Altiok E. Sweet pomegranate pekmez production: Effect of different production methods and activated carbon treatment on some quality parameters and 5‐hydroxymethyl furfural formation. J Food Process Preserv [Internet]. 2022 Feb 28;46(2):e16281. Available from: .
33. Özkan K, Türkmenoğlu G, Fakir H, Sağdıç O. Andız (Juniperus drupacea) pekmezinin üretimi, çeşitli fizikokimyasal ve biyoaktif özellikleri. Eur J Sci Technol [Internet]. 2022 Dec 24;46:74–81. Available from: .
34. Kuşçu A, Bulantekin Ö. The effects of production methods and storage on the chemical constituents of apple pekmez. J Food Sci Technol [Internet]. 2016 Jul 14;53(7):3083–92. Available from: .
35. Korkmaz A. The effects of different concentration methods on the chemical composition, functional and sensory attributes of molasses produced from grape (Vitis vinifera L.) juice. J Food Meas Charact [Internet]. 2023 Apr 13;17(2):2016–28. Available from: .
36. Şen L, Altiok D, Apaydin E. Improved qualitative properties of pear pekmez by natural zeolite treatment during production process. J Food Process Eng [Internet]. 2020 Oct 17;43(10). Available from: .
37. Abdul Rani MAA Bin, Karim NA, Kamarudin SK. Recent reaction systems for the synthesis of 5‐hydroxymethylfurfural (HMF) from carbohydrates with process development analysis: A review. Int J Energy Res [Internet]. 2022 Oct 25;46(13):18996–9050. Available from: .
38. Liu Z, Zhu L, Hu C. High-efficiency synthesis of 5-hydroxymethylfurfural from fructose over highly sulfonated organocatalyst. Ind Eng Chem Res [Internet]. 2020 Sep 30;59(39):17218–27. Available from: .
39. Orfanakis E, Vlasiadi M, Philippidis A, Zoumi A, Velegrakis M. Rapid method for the assessment of hydroxymethylfurfural (HMF) levels in honey by absorption spectroscopy. Food Sci Eng [Internet]. 2023 Dec 22;26–33. Available from: .
40. Zuo M, Wang X, Wang Q, Zeng X, Lin L. Aqueous‐natural deep eutectic solvent‐enhanced 5‐hydroxymethylfurfural production from glucose, starch, and food wastes. ChemSusChem [Internet]. 2022 Jul 7;15(13):e202101889. Available from: .

Details

Primary Language

English

Subjects

Natural Products and Bioactive Compounds

Journal Section

Research Article

Authors

Pınar Karagül
0000-0002-2569-776X
Türkiye

Elif Sine Düvenci ^*
0000-0003-1177-0978
Türkiye

Bora Karagül
0000-0002-0834-6183
Türkiye

Publication Date

March 7, 2026

Submission Date

October 8, 2025

Acceptance Date

January 28, 2026

Published in Issue

Year 2026 Number: 2026-1

DOI

https://doi.org/10.18596/jotcsa.1799306

IZ

https://izlik.org/JA25GG56DX

Cite

RIS / Bibtex

APA

Karagül, P., Düvenci, E. S., & Karagül, B. (2026). 5-Hydroxymethyl Furfural (HMF) Evaluation in Molasses Produced through Different Traditional Methods Using HPLC. Journal of the Turkish Chemical Society Section A: Chemistry, 2026-1, 1-11. https://doi.org/10.18596/jotcsa.1799306

AMA

1.Karagül P, Düvenci ES, Karagül B. 5-Hydroxymethyl Furfural (HMF) Evaluation in Molasses Produced through Different Traditional Methods Using HPLC. JOTCSA. 2026;(2026-1):1-11. doi:10.18596/jotcsa.1799306

Chicago

Karagül, Pınar, Elif Sine Düvenci, and Bora Karagül. 2026. “5-Hydroxymethyl Furfural (HMF) Evaluation in Molasses Produced through Different Traditional Methods Using HPLC”. Journal of the Turkish Chemical Society Section A: Chemistry, no. 2026-1: 1-11. https://doi.org/10.18596/jotcsa.1799306.

EndNote

Karagül P, Düvenci ES, Karagül B (March 1, 2026) 5-Hydroxymethyl Furfural (HMF) Evaluation in Molasses Produced through Different Traditional Methods Using HPLC. Journal of the Turkish Chemical Society Section A: Chemistry 2026-1 1–11.

IEEE

[1]P. Karagül, E. S. Düvenci, and B. Karagül, “5-Hydroxymethyl Furfural (HMF) Evaluation in Molasses Produced through Different Traditional Methods Using HPLC”, JOTCSA, no. 2026-1, pp. 1–11, Mar. 2026, doi: 10.18596/jotcsa.1799306.

ISNAD

Karagül, Pınar - Düvenci, Elif Sine - Karagül, Bora. “5-Hydroxymethyl Furfural (HMF) Evaluation in Molasses Produced through Different Traditional Methods Using HPLC”. Journal of the Turkish Chemical Society Section A: Chemistry. 2026-1 (March 1, 2026): 1-11. https://doi.org/10.18596/jotcsa.1799306.

JAMA

1.Karagül P, Düvenci ES, Karagül B. 5-Hydroxymethyl Furfural (HMF) Evaluation in Molasses Produced through Different Traditional Methods Using HPLC. JOTCSA. 2026;:1–11.

MLA

Karagül, Pınar, et al. “5-Hydroxymethyl Furfural (HMF) Evaluation in Molasses Produced through Different Traditional Methods Using HPLC”. Journal of the Turkish Chemical Society Section A: Chemistry, no. 2026-1, Mar. 2026, pp. 1-11, doi:10.18596/jotcsa.1799306.

Vancouver

1.Pınar Karagül, Elif Sine Düvenci, Bora Karagül. 5-Hydroxymethyl Furfural (HMF) Evaluation in Molasses Produced through Different Traditional Methods Using HPLC. JOTCSA. 2026 Mar. 1;(2026-1):1-11. doi:10.18596/jotcsa.1799306