Usability of the phenolic profile analysis method developed in RP-HPLC-PDA in natural products

Year 2024, Volume: 7 Issue: 1, 14 - 27, 30.06.2024

Yakup Kara , Ceren Birinci

https://doi.org/10.35206/jan.1430767

Abstract

The predominant majority of bioactive compounds in natural products are polyphenols. Reverse Phase High Performance Liquid Chromatography (HPLC) is the most employed analytical method for determining the polyphenol profiles of natural products. Analyses are conducted based on methods validated according to the number and type of phenolic standards used. In this study, it was aimed to determine 26 phenolic compound standards with HPLC-fotodiot array (PDA) detector, which is preferred for the separation of secondary metabolites commonly found in natural products. The analysis was carried out utilizing a C18 column (250 mm x 4.6 mm, 5 μm; GL Sciences) with a gradient program. The HPLC method was developed, determining the limit of detection within the range of 0.019-0.072 μg/mL, and the limit of quantification within the range of 0.063-0.239 μg/mL. All calibration curves exhibited linear corelations with R² values exceeding 0.994 across the specified range. The developed method has been optimized and validated by assessing detection and quantification limits, accuracy, repeatability, and recovery data suitable for phenolic analysis. It has been concluded that the optimized method allows for the rapid and reliable evaluation of the phenolic content of natural products and their quantitative determination.

Keywords

Phenolic content, Bee products, Plant, HPLC

RP-HPLC-PDA'da geliştirilen fenolik profil analiz yönteminin doğal ürünlerde kullanılabilirliği

Abstract

References

• Abdulla, K. K., Taha, E. M., & Rahim, S. M. (2014). Phenolic profile, antioxidant, and antibacterial effects of ethanol and aqueous extracts of Rheum ribes L. roots. Der Pharmacia Lettre, 6(5), 201-205.
• Ali, A. M., & Kunugi, H. (2021). Propolis, bee honey, and their components protect against coronavirus disease 2019 (COVID-19): A review of in silico, in vitro, and clinical studies. Molecules, 26(5), 1232.
• Bae, I. K., Ham, H. M., Jeong, M. H., Kim, D. H., & Kim, H. J. (2015). Simultaneous determination of 15 phenolic compounds and caffeine in teas and mate using RP-HPLC/UV detection: Method development and optimization of extraction process. Food chemistry, 172, 469-475.
• Benzie I. F. & Strain J. J. (1999). Ferric reducing/antioxidant power assay: direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. In Methods in enzymology Vol. 299, pp. 15–27. Academic Press.
• Can, Z., Kara, Y., Kolayli, S., & Çakmak, I. (2022). Antioxidant activity and phenolic composition of propolis from Marmara region, Türkiye. Journal of Apicultural Research, 1-7.
• Can, Z., Yildiz, O., Sahin, H., Turumtay, E. A., Silici, S., & Kolayli, S. (2015). An investigation of Turkish honeys: Their physico-chemical properties, antioxidant capacities and phenolic profiles. Food chemistry, 180, 133-141.
• Çayan, F., Deveci, E., Tel-Çayan, G., & Duru, M. E. (2020). Identification and quantification of phenolic acid compounds of twenty-six mushrooms by HPLC–DAD. Journal of Food Measurement and Characterization, 14, 1690-1698.
• Chaudhary, S. K., Sharma, K. C., Devi, S. I., Kar, A., Bhardwaj, P. K., Sharma, N., ... & Mukherjee, P. K. (2023). Evaluation of anti-HMG-CoA reductase potential and simultaneous determination of phenolic compounds in hydroalcoholic extract of Ficus cunia fruits by RP-HPLC. South African Journal of Botany, 155, 27-34.
• Fukumoto LR, Mazza G (2000) Assessing antioxidant and prooxidant activities of phenolic compounds. J of Agricultural and Food Chem 48(8):3597–3604
• Halagarda, M., Groth, S., Popek, S., Rohn, S., & Pedan, V. (2020). Antioxidant activity and phenolic profile of selected organic and conventional honeys from Poland. Antioxidants, 9(1), 44.
• Ignat, I., Volf, I., & Popa, V. I. (2011). A critical review of methods for characterisation of polyphenolic compounds in fruits and vegetables. Food chemistry, 126(4), 1821-1835.
• Kara, Y., Can, Z., & Kolaylı, S. (2022). Applicability of phenolic profile analysis method developed with RP-HPLC-PDA to some bee product. Brazilian Archives of Biology and Technology, 65.
• Karaçelik, A. A., & Şahin, H. (2021). Determination of chemical compositions, antioxidant and enzyme inhibitory activities of naturally growing Chenopodium album subsp. iranicum Aellen. Journal of the Institute of Science and Technology, 11(3), 2091-2101.
• Keser, S., Keser, F., Karatepe, M., Kaygili, O., Tekin, S., Turkoglu, I., ... & Sandal, S. (2020). Bioactive contents, in vitro antiradical, antimicrobial and cytotoxic properties of rhubarb (Rheum ribes L.) extracts. Natural product research, 34(23), 3353-3357.
• Kumar, N., & Goel, N. (2019). Phenolic acids: Natural versatile molecules with promising therapeutic applications. Biotechnology reports, 24, e00370.
• Kumar, N., Gupta, S., Chand Yadav, T., Pruthi, V., Kumar Varadwaj, P., & Goel, N. (2019). Extrapolation of phenolic compounds as multi-target agents against cancer and inflammation. Journal of Biomolecular Structure and Dynamics, 37(9), 2355-2369.
• Kumazawa, S., Hamasaka, T., & Nakayama, T. (2004). Antioxidant activity of propolis of various geographic origins. Food chemistry, 84(3), 329-339.
• Liu, Q., Cai, W., & Shao, X. (2008). Determination of seven polyphenols in water by high performance liquid chromatography combined with preconcentration. Talanta, 77(2), 679-683.
• Madrera, R. R., & Valles, B. S. (2020). Development and validation of ultrasound assisted extraction (UAE) and HPLC-DAD method for determination of polyphenols in dry beans (Phaseolus vulgaris). Journal of Food Composition and Analysis, 85, 103334.
• Meral, R. (2017). The effect of different temperatures on antioxidant activity and phenolic profile of the Rheum ribes. Yuzuncu Yıl University Journal of Agricultural Sciences, 27(1), 88-94.
• Michalaki, A., Karantonis, H. C., Kritikou, A. S., Thomaidis, N. S., & Dasenaki, M. E. (2023). Ultrasound-assisted extraction of total phenolic compounds and antioxidant activity evaluation from Oregano (Origanum vulgare ssp. hirtum) using response surface methodology and identification of specific phenolic compounds with HPLC-PDA and Q-TOF-MS/MS. Molecules, 28(5), 2033.
• Mısır, S., Özbek, M., & Hepokur, C. (2023). Bioactive Composition, Antioxidant, And Cytotoxic Activities of Rheum ribes Extracts: Rheum ribes ekstraktlarının çeşitli biyolojik aktiviteleri. Turkish Journal of Agriculture-Food Science and Technology, 11(8), 1345-1350.
• Molyneux P (2004) The use of the stable free radical diphenylpicrylhydrazyl (DPPH) for estimating antioxidant activity. Songklanakarin J Sci Technol 26(2):211–219.
• Oktay, M., Yildirim, A., Bilaloglu, V., & Gülçin, I. (2007). Antioxidant activity of different parts of isgin (Rheum ribes L.). Asian journal of chemistry, 19(4), 3047.
• Ożarowski, M., & Karpiński, T. M. (2023). The Effects of Propolis on Viral Respiratory Diseases. Molecules, 28(1), 359.
• Rashmi, H. B., & Negi, P. S. (2020). Phenolic acids from vegetables: A review on processing stability and health benefits. Food Research International, 136, 109298.
• Ribani, M., Collins, C. H., & Bottoli, C. B. (2007). Validation of chromatographic methods: Evaluation of detection and quantification limits in the determination of impurities in omeprazole. Journal of Chromatography A, 1156(1-2), 201-205.
• Santos, W. N. D., & MagalhÃes, B. E. D. (2020). Phenolic content and antioxidant capacity of infusions herbs: Optimization of phenolic extraction and HPLC-DAD method. Anais da Academia Brasileira de Ciências, 92.
• Seal, T. (2016). Quantitative HPLC analysis of phenolic acids, flavonoids and ascorbic acid in four different solvent extracts of two wild edible leaves, Sonchus arvensis and Oenanthe linearis of North-Eastern region in India. Journal of Applied Pharmaceutical Science, 6(2), 157-166.
• Skendi, A., Irakli, M., & Chatzopoulou, P. (2017). Analysis of phenolic compounds in Greek plants of Lamiaceae family by HPLC. Journal of applied research on medicinal and aromatic plants, 6, 62-69.
• Slinkard K, Singleton VL (1977) Total phenol analysis: automation and comparison with manual methods. Am J of Enology and Viticulture 28(1):49–55
• Sobral, F., Calhelha, R. C., Barros, L., Dueñas, M., Tomás, A., Santos-Buelga, C., ... & Ferreira, I. C. (2017). Flavonoid composition and antitumor activity of bee bread collected in northeast Portugal. Molecules, 22(2), 248.
• Švecová, B., Bordovská, M., Kalvachová, D., & Hájek, T. (2015). Analysis of Czech meads: Sugar content, organic acids content and selected phenolic compounds content. Journal of Food Composition and Analysis, 38, 80-88.
• Zhang, Y., Cao, C., Yang, Z., Jia, G., Liu, X., Li, X., ... & Li, A. (2023). Simultaneous determination of 20 phenolic compounds in propolis by HPLC-UV and HPLC-MS/MS. Journal of Food Composition and Analysis, 115, 104877.