HPLC-ELSD Kullanılarak Çeşitli Hasat Sonrası Klimakterik Meyvelerin Şeker İçeriklerinin Belirlenmesi; Farklı Olgunlaşma Aşamalarında Değerlendirme ve Yöntemin Analitik Eko-ölçek, NEMI, GAPI ve AGREE ile Yeşillik Profilinin Değerlendirilmesi

Öz

Meyvelerdeki ana şeker içeriği diyet açısından çok önemlidir. Bu çalışmada, HPLC-ELSD kullanılarak domates ve muzda farklı olgunlaşma aşamalarında (yeşil, olgunlaşma, olgun ve aşırı olgun) fruktoz, glukoz ve sukroz miktarlarındaki ve toplam şeker içeriğindeki değişiklikler araştırılmıştır. Kromatografik ayırmalar Phenomenex Luna NH2 sabit fazlı (5 μm partikül boyutu, 250 mm x 4,6 mm iç çap, 100 Å) bir HPLC kolonu kullanılarak gerçekleştirilmiş ve asetonitril:su (78:22, v/v) içeren bir mobil faz ile elüe edilmiştir. ELSD sistemi 40°C'lik bir sürüklenme tüpü sıcaklığında çalıştırılmış ve 350 kPa'lık bir nitrojen basıncı kullanılmıştır. Örneklerin şeker içeriği analiz edildiğinde, domatesler olgunlaştıkça sakkaroz seviyeleri azalırken, fruktoz ve glukoz miktarı artmıştır. Muz örneklerinde ise olgunlaşma döneminde fruktoz ve glukoz miktarı artarken sakkaroz içeriğinin önce arttığı sonra azaldığı bildirilmiştir. Bu sonuçlar, meyve olgunlaşması sırasında nişastanın sakkaroza, sakkarozun da fruktoz ve glukoza dönüştüğünü göstermektedir. Ayrıca, uygulanan HPLC-ELSD yönteminin klimakterik meyvelerin şeker içeriği açısından analiz edilmesinde kolaylıkla uygulanabilir olduğu gösterilmiştir. Ayrıca, uygulanan HPLC-ELSD yönteminin yeşillik profili Analitik Eko Ölçek, NEMI, GAPI ve AGREE araçları kullanılarak değerlendirilmiştir.

Anahtar Kelimeler

• klimakterik meyveler
• şekerler
• HPLC-ELSD
• domates
• muz
• yeşillik değerlendirmesi

Determination of Sugar Contents of Various Postharvest Climacteric Fruits Using HPLC-ELSD; Evaluation at Different Ripening Stages and Assessment of the Greenness Profile of the Method with Analytical Eco-scale, NEMI, GAPI and AGREE

Öz

The major sugar content in fruits is crucial in terms of diet. In this study, changes in fructose, glucose and sucrose amounts and total sugar content were investigated at different ripening stages (green, ripening, ripe and over-ripe) in tomatoes and bananas using an HPLC-ELSD method. Chromatographic separations were conducted using an HPLC column with a Phenomenex Luna NH2 stationary phase (5 μm particle size, 250 mm x 4.6 mm internal diameter, 100 Å), and eluted with a mobile phase consisting of acetonitrile:water (78:22, v/v). Quantitative results demonstrated a significant increase (p < 0.0001) in fructose and glucose contents of bananas from 0.87 g/100 g and 1.27 g/100 g at the green stage to 5.76 g/100 g and 11.13 g/100 g at the over-ripe stage, respectively, while sucrose peaked at 4.41 g/100 g during the ripening stage and declined thereafter. In tomatoes, fructose and glucose contents showed a moderate but statistically significant rise (p < 0.01) from 4.21 g/100 g and 6.48 g/100 g to 4.86 g/100 g and 6.55 g/100 g, whereas sucrose became undetectable after the ripening stage. Total sugar content increased by approximately sixfold in bananas and slightly in tomatoes throughout ripening. These results indicate that during fruit ripening, sucrose is converted into fructose and glucose. The applied HPLC-ELSD method has been shown to be easily applicable for analysing climacteric fruits for sugar content. In addition, the greenness profile of the applied HPLC-ELSD method was evaluated using AES, NEMI, GAPI, and AGREE tools, confirming that the method is environmentally acceptable and consistent with the principles of Green Analytical Chemistry.

Anahtar Kelimeler

• climacteric fruits
• sugars
• HPLC-ELSD
• tomato
• banana
• greenness assessment

Kaynakça

1. Adão, R. C., & Glória, M. B. A. (2005). Bioactive amines and carbohydrate changes during ripening of “Prata” banana (Musa acuminata x M. balbisiana). Food Chemistry, 90(4), 705–711. https://doi.org/10.1016/J.FOODCHEM.2004.05.020
2. Adewale, I. O., Adefila, A., & Adewale, T. B. (2013). Changes in amylase activity, soluble sugars and proteins of unripe banana and plantain during ripening. Annual Review & Research in Biology, 34, 678-685.
3. Agius, C., von Tucher, S., Poppenberger, B., & Rozhon, W. (2018). Quantification of sugars and organic acids in tomato fruits. MethodsX, 5, 537–550. https://doi.org/10.1016/J.MEX.2018.05.014
4. Alferez, F., de Carvalho, D. U., & Boakye, D. (2021). Interplay between abscisic acid and gibberellins, as related to ethylene and sugars, in regulating maturation of non-climacteric fruit. International Journal of Molecular Sciences, 22(2), 669. https://doi.org/10.3390/ijms22020669
5. Chen, Y., Grimplet, J., David, K., Castellarin, S. D., Terol, J., Wong, D. C. J., Luo, Z., Schaffer, R., Celton, J. M., Talon, M., Gambetta, G. A., & Chervin, C. (2018). Ethylene receptors and related proteins in climacteric and non-climacteric fruits. Plant Science, 276, 63–72. https://doi.org/10.1016/J.PLANTSCI.2018.07.012
6. Filip, M., Vlassa, M., Coman, V., & Halmagyi, A. (2016). Simultaneous determination of glucose, fructose, sucrose and sorbitol in the leaf and fruit peel of different apple cultivars by the HPLC–RI optimized method. Food Chemistry, 199, 653–659. https://doi.org/10.1016/J.FOODCHEM.2015.12.060
7. Gałuszka, A., Migaszewski, Z. M., Konieczka, P., & Namieśnik, J. (2012). Analytical Eco-Scale for assessing the greenness of analytical procedures. TrAC Trends in Analytical Chemistry, 37, 61-72. https://doi.org/10.1016/j.trac.2012.03.013
8. Gao, Y., Tian, P., Li, J., Cao, Y., Xu, W., & Li, J. (2019). Transcriptional changes during tomato ripening and influence of brackish water irrigation on fruit transcriptome and sugar content. Plant Physiology and Biochemistry, 145, 21–33. https://doi.org/10.1016/J.PLAPHY.2019.10.025

9. Gautier, H., Diakou-Verdin, V., Bénard, C., Reich, M., Buret, M., Bourgaud, F., Poëssel, J. L., Caris-Veyrat, C., & Génard, M. (n.d.). How Does Tomato Quality (Sugar, Acid, and Nutritional Quality) Vary with Ripening Stage, Temperature, and Irradiance? Journal of Agricultural and Food Chemistry, 56(4), 1241-1250. https://doi.org/10.1021/jf072196t
10. Georgelis, N., Fencil, K., & Richael, C. M. (2018). Validation of a rapid and sensitive HPLC/MS method for measuring sucrose, fructose and glucose in plant tissues. Food Chemistry, 262, 191–198. https://doi.org/10.1016/J.FOODCHEM.2018.04.051
11. Grembecka, M., Lebiedzińska, A., & Szefer, P. (2014). Simultaneous separation and determination of erythritol, xylitol, sorbitol, mannitol, maltitol, fructose, glucose, sucrose and maltose in food products by high performance liquid chromatography coupled to charged aerosol detector. Microchemical Journal, 117, 77–82. https://doi.org/10.1016/J.MICROC.2014.06.012
12. Hussain, C.M., Hussain, G., & Keçili, R. (2025b). Disposable analytical chemistry: Design, application & sustainability. TrAC - Trends in Analytical Chemistry. https://doi.org/10.1016/j.trac.2025.118414.
13. Ivancic, T., Jakopic, J., Veberic, R., Vesel, V., & Hudina, M. (2022). Effect of Ripening on the Phenolic and Sugar Contents in the Meso- and Epicarp of Olive Fruits (Olea europaea L.) Cultivar ‘Leccino.’ Agriculture (Switzerland), 12(9), 1347. https://doi.org/10.3390/AGRICULTURE12091347/S1
14. Jaglan, P., Buttar, H. S., Al-bawareed, O. A., & Chibisov, S. (2022). Potential health benefits of selected fruits: apples, blueberries, grapes, guavas, mangos, pomegranates, and tomatoes. Functional Foods and Nutraceuticals in Metabolic and Non-Communicable Diseases, 359–370. https://doi.org/10.1016/B978-0-12-819815-5.00026-4
15. Kaur, D., Sharma, R., Wani, A. A., Gill, S., & Sogi, D. S. (2006). Physicochemical Changes in Seven Tomato (Lycopersicon esculentum) Cultivars During Ripening. International Journal of Food Properties, 9(4), 747–757. https://doi.org/10.1080/10942910600575716
16. Keith, L. H., Gron, L. U., & Young, J. L. (2007). Green analytical methodologies. Chemical reviews, 107(6), 2695-2708. https://doi.org/10.1021/cr068359e
17. Kim, H. Y., Farcuh, M., Cohen, Y., Crisosto, C., Sadka, A., & Blumwald, E. (2015). Non-climacteric ripening and sorbitol homeostasis in plum fruits. Plant Science, 231, 30–39. https://doi.org/10.1016/J.PLANTSCI.2014.11.002
18. Klee, H., genetics, J. G.-A. review of, & 2011, undefined. (2011). Genetics and control of tomato fruit ripening and quality attributes. Annual Reviews, 45, 41–59. https://doi.org/10.1146/annurev-genet-110410-132507
19. Kou, X., Feng, Y., Yuan, S., Zhao, X., Wu, C., Wang, C., & Xue, Z. (2021). Different regulatory mechanisms of plant hormones in the ripening of climacteric and non-climacteric fruits: a review. Plant Molecular Biology, 1-21. https://doi.org/10.1007/s11103-021-01199-9
20. Kou, X., & Wu, M. (2018). Characterization of climacteric and non-climacteric fruit ripening. Methods in Molecular Biology, 1744, 89–102. https://doi.org/10.1007/978-1-4939-7672-0_7/COVER
21. Liu, R. H. (2003). Health benefits of fruit and vegetables are from additive and synergistic combinations of phytochemicals. American Journal of Clinical Nutrition, 78 (3). https://doi.org/10.1093/AJCN/78.3.517S
22. Ma, C., Sun, Z., Chen, C., Zhang, L., & Zhu, S. (2014). Simultaneous separation and determination of fructose, sorbitol, glucose and sucrose in fruits by HPLC–ELSD. Food Chemistry, 145, 784–788. https://doi.org/10.1016/J.FOODCHEM.2013.08.135
23. Maduwanthi, S. D. T., & Marapana, R. A. U. J. (2017). Biochemical changes during ripening of banana: A review. International Journal of Food Science and Nutrition, Vol.2 (5), pp. 166-169. http://dr.lib.sjp.ac.lk/handle/123456789/7777
24. Mojsiewicz-Pieńkowska, K. (2009). On the issue of characteristic evaporative light scattering detector response. Critical Reviews in Analytical Chemistry, 39(2), 89–94. https://doi.org/10.1080/15389580802570218
25. Nogueira, L. C., Silva, F., Ferreira, I. M. P. L. V. O., & Trugo, L. C. (2005). Separation and quantification of beer carbohydrates by high-performance liquid chromatography with evaporative light scattering detection. Journal of Chromatography A, 1065(2), 207–210. https://doi.org/10.1016/J.CHROMA.2004.12.074
26. Payasi, A., Biochemistry, G. S.-J. of food, & 2010, undefined. (2010). Ripening of climacteric fruits and their control. Journal of food Biochemistry, 34(4), 679-710. https://doi.org/10.1111/j.1745-4514.2009.00307.x
27. Pena-Pereira, F., Wojnowski, W., & Tobiszewski, M. (2020). AGREE—Analytical GREEnness metric approach and software. Analytical chemistry, 92(14), 10076-10082. https://doi.org/10.1021/acs.analchem.0c01887
28. Płotka-Wasylka, J. (2018). A new tool for the evaluation of the analytical procedure: Green Analytical Procedure Index. Talanta, 181, 204-209. https://doi.org/10.1016/j.talanta.2018.01.013
29. Ruiz-jimé Nez, J., Priego-capote, F., & Dolores Luque De Castro, M. A. (2010). Qualitative and Quantitative Sugar Profiling in Olive Fruits, Leaves, and Stems by Gas Chromatography-Tandem Mass Spectrometry (GC-MS/MS) after Ultrasound-Assisted Leaching. J. Agric. Food Chem, 58, 12292–12299. https://doi.org/10.1021/jf102350s
30. Singh, B., Singh, J. P., Kaur, A., & Singh, N. (2016). Bioactive compounds in banana and their associated health benefits - A review. Food Chemistry, 206, 1–11. https://doi.org/10.1016/J.FOODCHEM.2016.03.033
31. Sezgin, B., Arli, G., & Soyseven, M. (2025). Multi-metric greenness, performance and applicability evaluation of a green HPLC-ELSD method for polysorbate 80 determination in various pharmaceuticals compared with AI-assisted scoring systems. Microchemical Journal, 218. https://doi.org/10.1016/j.microc.2025.115710.
32. Soyseven, M., Sezgin, B., Acar, E., & Arli, G. (2023). Green, Rapid, Robust Determination of Aspartame in Beverages by High-Performance Liquid Chromatography-Evaporative Light Scattering Detection (HPLC-ELSD): Method Development and Validation. Analytical Letters, 56(11), 1773–1783. https://doi.org/10.1080/00032719.2022.2146129
33. Soyseven, M., Sezgin, B., & Arli, G. (2022). A novel, rapid and robust HPLC-ELSD method for simultaneous determination of fructose, glucose and sucrose in various food samples: Method development and validation. Journal of Food Composition and Analysis, 107. https://doi.org/10.1016/J.JFCA.2022.104400
34. Subedi, P. P., & Walsh, K. B. (2011). Assessment of sugar and starch in intact banana and mango fruit by SWNIR spectroscopy. Postharvest Biology and Technology, 62(3), 238–245. https://doi.org/10.1016/J.POSTHARVBIO.2011.06.014
35. Tsai, Y. H., Tsai, C. W., & Tipple, C. A. (2022). A validated method for the analysis of sugars and sugar alcohols related to explosives via liquid chromatography mass spectrometry (LC-MS) with post-column addition. Forensic Chemistry, 28. https://doi.org/10.1016/J.FORC.2022.100404
36. Yumbya, P., Ambuko, J., Hutchinson, M., Owino, W., Juma, J., Machuka, E., & Mutuku, J. M. (2021). Transcriptome analysis to elucidate hexanal’s mode of action in preserving the post-harvest shelf life and quality of banana fruits (Musa acuminata). Journal of Agriculture and Food Research, 3. https://doi.org/10.1016/J.JAFR.2021.100114