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Kağıt Tabanlı Kolorimetrik Yöntemle Askorbik Asit (Vitamin C) Tayini

Year 2022, , 597 - 606, 30.06.2022
https://doi.org/10.35414/akufemubid.1015952

Abstract

Askorbik asit (AA), birçok yiyecek ve içecekte C vitamini olarak bilinen ve antioksidan özelliği olan en önemli suda çözünür organik biyomoleküldür. Gıdaların antioksidan özelliklerinin belirlenmesine yönelik geliştirilen farklı yöntemlerin sayısı giderek artmaktadır. Kağıt tabanlı mikroakışkan analitik cihazlar (µPAD'ler), son yıllarda hızlı, basit ve uygun maliyetli saha analizlerinin geliştirilmesinde uygulanmaktadır. Geliştirilen bu analizler, laboratuar donanımlı kalitatif ve kantitatif analizlere alternatif olarak kullanılmaktadır. Bu çalışma kapsamında, askorbik asit tayini için kağıt tabanlı kolorimetrik bir yöntemin uygulaması yapılmıştır. Whatman filtre kağıtları üzerinde vaks kalem yardımıyla hidrofobik bariyerli geometrik şekiller oluşturulmuştur. Bu şekillerin içerisinde AgNO3 çözeltisinin antioksidan varlığında indirgenmesi prensibi kullanılarak meydana gelen renk dönüşümlerinin görüntüleri akıllı telefon kamerası yardımıyla kaydedilmiştir. Standart AA çözeltilerinin konsantrasyonlarının tespiti için ImageJ yazılımı, görüntü işleme prosesinde kullanılmıştır. 0,2 ile 1,0 mM aralığında AA konsantrasyonları için doğrusal kalibrasyon eğrileri gri ölçek ve RGB (Kırmızı-Yeşil-Mavi) değerlerinde elde edilmiştir. Kalibrasyon eğrilerinin determinasyon katsayısı (R2) sırasıyla; gri ölçek için 0,9389, Kırmızı, Yeşil ve Mavi kanallar için 0,9728, 0,9628 ve 0,8859 olarak hesaplanmıştır. Yüksek Performanslı Sıvı Kromatografisi (HPLC) standart bir yöntem olarak kağıt tabanlı yöntem ile karşılaştırmak için kullanılmıştır. Gerçek gıda örneğinde askorbik asit tayini için taze sıkılmış portakal suları kullanılmıştır. HPLC yöntemiyle kolorimetrik yöntem sonucu portakal sularında belirlenen askorbik asit değerleri istatistiksel olarak karşılaştırılmıştır.

Supporting Institution

Ardahan Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü

Project Number

BAP2018-027

Thanks

Bu çalışma Ardahan Üniversitesi Bilimsel Araştırma Projeleri Kordinatörlüğü tarafından BAP 2018-027 No’lu Proje kapsamında desteklenmiştir.

References

  • Busa, L. S. A., S. Mohammadi, M. Maeki, A. Ishida, H. Tani and M. Tokeshi 2016. Advances in Microfluidic Paper-Based Analytical Devices for Food and Water Analysis. Micromachines 7(5), 86.
  • Carr, A. C. and M. C. M. Vissers 2013. Synthetic or Food-Derived Vitamin C-Are They Equally Bioavailable? Nutrients 5(11), 4284-4304.
  • de Quirós, A. Rodríguez-Bernaldo, M Fernández-Arias, and J López-Hernández 2009. A screening method for the determination of ascorbic acid in fruit juices and soft drinks. Food chemistry 116(2), 509-512.
  • Dennison, D. B., T. G. Brawley and G. L. Hunter 1981. Rapid high-performance liquid chromatographic determination of ascorbic acid and combined ascorbic acid-dehydroascorbic acid in beverages. Journal of Agricultural and Food Chemistry 29(5), 927-929.
  • Güçlü, K., K. Sözgen, E. Tütem, M. Özyürek and R. Apak 2005. Spectrophotometric determination of ascorbic acid using copper (II)–neocuproine reagent in beverages and pharmaceuticals. Talanta 65(5), 1226-1232.
  • Hawkins, K. R. and B. H. Weigl 2010. Microfluidic diagnostics for low-resource settings. Microfluidics, bioMEMs, and medical microsystems VIII, International Society for Optics and Photonics.
  • Hu, J., S. Q. Wang, L. Wang, F. Li, B. Pingguan-Murphy, T. J. Lu and F. Xu 2014. Advances in paper-based point-of-care diagnostics. Biosensors & Bioelectronics 54, 585-597.
  • Hua, M. Z., S. M. Li, S. Wang and X. N. Lu 2018. Detecting Chemical Hazards in Foods Using Microfluidic Paper-Based Analytical Devices (PADs): The Real-World Application. Micromachines 9(1).
  • Jeeva, J. S., J. Sunitha, R. Ananthalakshmi, S. Rajkumari, M. Ramesh and R. Krishnan 2015. Enzymatic antioxidants and its role in oral diseases. Journal of Pharmacy & Bioallied Sciences 7(Suppl 2), S331-S333.
  • Kapur, A., A. Hasković, A. Čopra-Janićijević, L. Klepo, A. Topčagić, I. Tahirović and E. Sofić 2012. Spectrophotometric analysis of total ascorbic acid content in various fruits and vegetables. Bulletin of the Chemists and Technologists of Bosnia and Herzegovina 38(4), 39-42.
  • Lykkesfeldt, J., S. Loft and H. E. Poulsen 1995. Determination of ascorbic acid and dehydroascorbic acid in plasma by high-performance liquid chromatography with coulometric detection-are they reliable biomarkers of oxidative stress? Analytical biochemistry 229(2), 329-335.
  • Lykkesfeldt, J. and P. Tveden-Nyborg 2019. The Pharmacokinetics of Vitamin C. Nutrients 11(10).
  • Mabey, D., R. W. Peeling, A. Ustianowski and M. D. Perkins 2004. Diagnostics for the developing world. Nature Reviews Microbiology 2(3), 231-240.
  • Maritim, A. C., R. A. Sanders and J. B. Watkins, 3rd 2003. Diabetes, oxidative stress, and antioxidants: a review. J Biochem Mol Toxicol 17(1), 24-38.
  • Martinez, A. W., S. T. Phillips, G. M. Whitesides and E. Carrilho 2010. Diagnostics for the Developing World: Microfluidic Paper-Based Analytical Devices. Analytical Chemistry 82(1), 3-10.
  • Pinchuk, I., H. Shoval, Y. Dotan and D. Lichtenberg 2012. Evaluation of antioxidants: scope, limitations and relevance of assays. Chem Phys Lipids 165(6), 638-647.
  • Pisoschi, A. M., A. Pop, A. I. Serban and C. Fafaneata 2014. Electrochemical methods for ascorbic acid determination. Electrochimica Acta 121, 443-460.
  • Ratnam, D. V., D. D. Ankola, V. Bhardwaj, D. K. Sahana and M. N. Kumar 2006. Role of antioxidants in prophylaxis and therapy: A pharmaceutical perspective. J Control Release 113(3), 189-207.
  • Uğur, H., S. Eker, J. Çatak and M. Yaman 2020. Vitamin C ve Hastalıklar Üzerine Etkisi. Avrupa Bilim ve Teknoloji Dergisi 19(19), 746-756.
  • Washko, P. W., W. O. Hartzell and M. Levine 1989. Ascorbic acid analysis using high-performance liquid chromatography with coulometric electrochemical detection. Analytical biochemistry 181(2), 276-282.
  • Washko, P. W., R. W. Welch, K. R. Dhariwal, Y. Wang and M. Levine 1992. Ascorbic acid and dehydroascorbic acid analyses in biological samples. Analytical biochemistry 204(1), 1-14.
  • Whitesides, G. M. 2006. The origins and the future of microfluidics. Nature 442(7101), 368-373.

Determination of Ascorbic Acid (Vitamin C) with Paper Based Colorimetric Method

Year 2022, , 597 - 606, 30.06.2022
https://doi.org/10.35414/akufemubid.1015952

Abstract

Ascorbic acid (AA) is the most important water-soluble organic biomolecule with antioxidant properties, known as vitamin C in many foods and beverages. The number of different methods developed for the determination of antioxidant properties of foods is increasing. Paper-based microfluidic analytical devices (µPADs) have been applied in the development of fast, simple, and cost-effective field analyzes in recent years. These developed analyzes are used as an alternative to laboratory-equipped qualitative and quantitative analyzes. In this study, a paper-based colorimetric method was applied for the determination of ascorbic acid. Geometric shapes with hydrophobic barrier were created on Whatman filter papers with the help of a wax pen. Using the principle of reduction of the AgNO3 solution in the presence of antioxidants in these shapes, the images of the color transformations were recorded with the help of a smartphone camera. ImageJ software was used in the image processing to determine the concentrations of standard AA solutions. Linear calibration curves for AA concentrations in the 0,2 to 1,0 mM range were obtained in grayscale and RGB (Red-Green-Blue) values. The coefficient of determination of the calibration curves (R2), respectively; 0,9389 for grayscale, 0,9728, 0,9628, and 0,8859 for Red, Green, and Blue channels. High-Performance Liquid Chromatography (HPLC) was used as a standard method to compare with the paper-based method. Freshly squeezed orange juice was used for the determination of ascorbic acid in the real food sample. The ascorbic acid value determined in orange juices by HPLC and colorimetric methods were statistically compared.

Project Number

BAP2018-027

References

  • Busa, L. S. A., S. Mohammadi, M. Maeki, A. Ishida, H. Tani and M. Tokeshi 2016. Advances in Microfluidic Paper-Based Analytical Devices for Food and Water Analysis. Micromachines 7(5), 86.
  • Carr, A. C. and M. C. M. Vissers 2013. Synthetic or Food-Derived Vitamin C-Are They Equally Bioavailable? Nutrients 5(11), 4284-4304.
  • de Quirós, A. Rodríguez-Bernaldo, M Fernández-Arias, and J López-Hernández 2009. A screening method for the determination of ascorbic acid in fruit juices and soft drinks. Food chemistry 116(2), 509-512.
  • Dennison, D. B., T. G. Brawley and G. L. Hunter 1981. Rapid high-performance liquid chromatographic determination of ascorbic acid and combined ascorbic acid-dehydroascorbic acid in beverages. Journal of Agricultural and Food Chemistry 29(5), 927-929.
  • Güçlü, K., K. Sözgen, E. Tütem, M. Özyürek and R. Apak 2005. Spectrophotometric determination of ascorbic acid using copper (II)–neocuproine reagent in beverages and pharmaceuticals. Talanta 65(5), 1226-1232.
  • Hawkins, K. R. and B. H. Weigl 2010. Microfluidic diagnostics for low-resource settings. Microfluidics, bioMEMs, and medical microsystems VIII, International Society for Optics and Photonics.
  • Hu, J., S. Q. Wang, L. Wang, F. Li, B. Pingguan-Murphy, T. J. Lu and F. Xu 2014. Advances in paper-based point-of-care diagnostics. Biosensors & Bioelectronics 54, 585-597.
  • Hua, M. Z., S. M. Li, S. Wang and X. N. Lu 2018. Detecting Chemical Hazards in Foods Using Microfluidic Paper-Based Analytical Devices (PADs): The Real-World Application. Micromachines 9(1).
  • Jeeva, J. S., J. Sunitha, R. Ananthalakshmi, S. Rajkumari, M. Ramesh and R. Krishnan 2015. Enzymatic antioxidants and its role in oral diseases. Journal of Pharmacy & Bioallied Sciences 7(Suppl 2), S331-S333.
  • Kapur, A., A. Hasković, A. Čopra-Janićijević, L. Klepo, A. Topčagić, I. Tahirović and E. Sofić 2012. Spectrophotometric analysis of total ascorbic acid content in various fruits and vegetables. Bulletin of the Chemists and Technologists of Bosnia and Herzegovina 38(4), 39-42.
  • Lykkesfeldt, J., S. Loft and H. E. Poulsen 1995. Determination of ascorbic acid and dehydroascorbic acid in plasma by high-performance liquid chromatography with coulometric detection-are they reliable biomarkers of oxidative stress? Analytical biochemistry 229(2), 329-335.
  • Lykkesfeldt, J. and P. Tveden-Nyborg 2019. The Pharmacokinetics of Vitamin C. Nutrients 11(10).
  • Mabey, D., R. W. Peeling, A. Ustianowski and M. D. Perkins 2004. Diagnostics for the developing world. Nature Reviews Microbiology 2(3), 231-240.
  • Maritim, A. C., R. A. Sanders and J. B. Watkins, 3rd 2003. Diabetes, oxidative stress, and antioxidants: a review. J Biochem Mol Toxicol 17(1), 24-38.
  • Martinez, A. W., S. T. Phillips, G. M. Whitesides and E. Carrilho 2010. Diagnostics for the Developing World: Microfluidic Paper-Based Analytical Devices. Analytical Chemistry 82(1), 3-10.
  • Pinchuk, I., H. Shoval, Y. Dotan and D. Lichtenberg 2012. Evaluation of antioxidants: scope, limitations and relevance of assays. Chem Phys Lipids 165(6), 638-647.
  • Pisoschi, A. M., A. Pop, A. I. Serban and C. Fafaneata 2014. Electrochemical methods for ascorbic acid determination. Electrochimica Acta 121, 443-460.
  • Ratnam, D. V., D. D. Ankola, V. Bhardwaj, D. K. Sahana and M. N. Kumar 2006. Role of antioxidants in prophylaxis and therapy: A pharmaceutical perspective. J Control Release 113(3), 189-207.
  • Uğur, H., S. Eker, J. Çatak and M. Yaman 2020. Vitamin C ve Hastalıklar Üzerine Etkisi. Avrupa Bilim ve Teknoloji Dergisi 19(19), 746-756.
  • Washko, P. W., W. O. Hartzell and M. Levine 1989. Ascorbic acid analysis using high-performance liquid chromatography with coulometric electrochemical detection. Analytical biochemistry 181(2), 276-282.
  • Washko, P. W., R. W. Welch, K. R. Dhariwal, Y. Wang and M. Levine 1992. Ascorbic acid and dehydroascorbic acid analyses in biological samples. Analytical biochemistry 204(1), 1-14.
  • Whitesides, G. M. 2006. The origins and the future of microfluidics. Nature 442(7101), 368-373.
There are 22 citations in total.

Details

Primary Language Turkish
Subjects Analytical Chemistry, Food Engineering, Nanotechnology
Journal Section Articles
Authors

Akif Göktuğ Bozkurt 0000-0002-2954-2132

Project Number BAP2018-027
Publication Date June 30, 2022
Submission Date October 28, 2021
Published in Issue Year 2022

Cite

APA Bozkurt, A. G. (2022). Kağıt Tabanlı Kolorimetrik Yöntemle Askorbik Asit (Vitamin C) Tayini. Afyon Kocatepe Üniversitesi Fen Ve Mühendislik Bilimleri Dergisi, 22(3), 597-606. https://doi.org/10.35414/akufemubid.1015952
AMA Bozkurt AG. Kağıt Tabanlı Kolorimetrik Yöntemle Askorbik Asit (Vitamin C) Tayini. Afyon Kocatepe Üniversitesi Fen Ve Mühendislik Bilimleri Dergisi. June 2022;22(3):597-606. doi:10.35414/akufemubid.1015952
Chicago Bozkurt, Akif Göktuğ. “Kağıt Tabanlı Kolorimetrik Yöntemle Askorbik Asit (Vitamin C) Tayini”. Afyon Kocatepe Üniversitesi Fen Ve Mühendislik Bilimleri Dergisi 22, no. 3 (June 2022): 597-606. https://doi.org/10.35414/akufemubid.1015952.
EndNote Bozkurt AG (June 1, 2022) Kağıt Tabanlı Kolorimetrik Yöntemle Askorbik Asit (Vitamin C) Tayini. Afyon Kocatepe Üniversitesi Fen Ve Mühendislik Bilimleri Dergisi 22 3 597–606.
IEEE A. G. Bozkurt, “Kağıt Tabanlı Kolorimetrik Yöntemle Askorbik Asit (Vitamin C) Tayini”, Afyon Kocatepe Üniversitesi Fen Ve Mühendislik Bilimleri Dergisi, vol. 22, no. 3, pp. 597–606, 2022, doi: 10.35414/akufemubid.1015952.
ISNAD Bozkurt, Akif Göktuğ. “Kağıt Tabanlı Kolorimetrik Yöntemle Askorbik Asit (Vitamin C) Tayini”. Afyon Kocatepe Üniversitesi Fen Ve Mühendislik Bilimleri Dergisi 22/3 (June 2022), 597-606. https://doi.org/10.35414/akufemubid.1015952.
JAMA Bozkurt AG. Kağıt Tabanlı Kolorimetrik Yöntemle Askorbik Asit (Vitamin C) Tayini. Afyon Kocatepe Üniversitesi Fen Ve Mühendislik Bilimleri Dergisi. 2022;22:597–606.
MLA Bozkurt, Akif Göktuğ. “Kağıt Tabanlı Kolorimetrik Yöntemle Askorbik Asit (Vitamin C) Tayini”. Afyon Kocatepe Üniversitesi Fen Ve Mühendislik Bilimleri Dergisi, vol. 22, no. 3, 2022, pp. 597-06, doi:10.35414/akufemubid.1015952.
Vancouver Bozkurt AG. Kağıt Tabanlı Kolorimetrik Yöntemle Askorbik Asit (Vitamin C) Tayini. Afyon Kocatepe Üniversitesi Fen Ve Mühendislik Bilimleri Dergisi. 2022;22(3):597-606.


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