Research Article
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Chlorogenic Acid: HPLC Quantification and In Vitro Assessment of Proliferative and Migration Effects on Human Dermal Fibroblast Cells

Year 2024, , 299 - 308, 30.06.2024

Abdulbaki Akpınar , Fatma Demirkaya Miloğlu , Gülşah Gündoğdu , Leyla Güven , Burak Bayrak , Yücel Kadıoğlu

https://doi.org/10.17776/csj.1440382

Abstract

Chlorogenic acid (CA) exhibits diverse biological activities, including antioxidant and antiinflammatory effects. This research aims to develop, optimize, and validate an HPLC method to quantify CA in methanol and investigate its in vitro proliferative and cell migration effects on human-dermal-fibroblast (HDF) cell lines in a dose-dependent manner. The HPLC experimental conditions were optimized using the central composite design (CCD) method for determining CA. Chromatographic separation occurred at a wavelength of 330 nm. Under the optimized conditions, the method exhibited linearity across a concentration range of 0.1-100 µg/mL, demonstrating sensitivity (LOQ:0.1µg/mL), precision (RSD%≤3.32), and accuracy (RE%≤4.05). To evaluate the in vitro proliferative and cell migration effects on HDFs, we employed the XTT cell proliferation assay and TAS-TOS commercial kits. The XTT assay revealed that CA displayed a proliferative effect within the concentration range of 75-250 µM (P <0.01), and at a concentration of 125 µM, TAS levels increased significantly (P<0.05). The scratch assay demonstrated that HDF cell migration increased at 12 h, with substantial closure of the wound area at 24 h when treated with CA concentrations between 75-125 µM. The results demonstrate that pure chlorogenic acid extracted from plants exhibits dose-dependent effects on cell proliferation, antioxidant, and cell migration

Keywords

Chlorogenic acid HPLC Experimental design Central composite design

References

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  • [3] Naveed M., Hejazi V., Abbas M., Kamboh A.A., Khan G.J., Shumzaid M., XiaoHui Z., Chlorogenic acid (CGA): A pharmacological review and call for further research, Biomedicine & pharmacotherapy., 97 (2018) 67-74.
  • [4] Budama-Kılınç Y., Klorojenik Asit Yüklü PLGA Nanopartiküllerinin Üretimi ve Antimikrobiyal Etkinliğinin Belirlenmesi, Türk Mikrobiyoloji Cemiyeti Dergisi., 49(1) (2019) 47-54.
  • [5] Feng R., Lu Y., Bowman L.L., Qian,Y., Castranova V., Ding M., Inhibition of activator protein-1, NF-κB, and MAPKs and induction of phase 2 detoxifying enzyme activity by chlorogenic acid, Journal of Biological Chemistry., 280 (2005) 27888-27895.
  • [6] Zhong C., Wall N.R., Zu Y., Sui G., Therapeutic application of natural medicine monomers in cancer treatment, Current Medicinal Chemistry., 24(34) (2017) 3681-3697.
  • [7] Bagdas D., Gul N.Y., Topal A., Tas S., Ozyigit M.O., Cinkilic N., Gurun M.S., Pharmacologic overview of systemic chlorogenic acid therapy on experimental wound healing, Naunyn-Schmiedeberg's archives of pharmacology., 387 (2014) 1101-1116.
  • [8] Chen W.C., Liou S.S., Tzeng T.F., Lee S.L., Liu I.M., Effect of topical application of chlorogenic acid on excision wound healing in rats, Planta medica., (2013) 616-621.
  • [9] Chaowuttikul C., Palanuvej C., Ruangrungsi N., Quantification of chlorogenic acid, rosmarinic acid, and caffeic acid contents in selected Thai medicinal plants using RP-HPLC-DAD, Brazilian Journal of Pharmaceutical Sciences., 56 (2020) e17547.
  • [10] Kolaylı S., Şahin H., Ulusoy E., Tarhan Ö., Phenolic composition and antioxidant capacities of Helichrysum plicatum, Hacettepe Journal of Biology and Chemistry., 38(4) (2010) 269-276.
  • [11] Albayrak S., Aksoy A., Sagdic O., Hamzaoglu E.,Compositions, antioxidant and antimicrobial activities of Helichrysum (Asteraceae) species collected from Turkey, Food chemistry., 119(1) (2010) 114-122.
  • [12] Albayrak S., Aksoy A., Sağdıç O., Budak Ü., Phenolic compounds and antioxidant and antimicrobial properties of Helichrysum species collected from eastern Anatolia, Turkey,Turkish Journal of Biology., 34(4) (2010) 463-473.
  • [13] Ayelign A., Sabally K., Determination of chlorogenic acids (CGA) in coffee beans using HPLC, American Journal of Research Communication., 1(2) (2013) 78-91.
  • [14] De Luca S., Ciotoli E., Biancolillo A., Bucci R., Magrì A.D.,Marini F., Simultaneous quantification of caffeine and chlorogenic acid in coffee green beans and varietal classification of the samples by HPLC-DAD coupled with chemometrics, Environmental Science and Pollution Research., 25 (2018) 28748-28759.
  • [15] Li H., Chen B., Yao S., Application of ultrasonic technique for extracting chlorogenic acid from Eucommia ulmodies Oliv.(E. ulmodies), Ultrasonics sonochemistry., 12(4) (2005) 295-300.
  • [16] Zhang B., Yang R., Liu C.Z., Microwave-assisted extraction of chlorogenic acid from flower buds of Lonicera japonica Thunb, Separation and Purification Technology., 62(2) (2008) 480-483.
  • [17] Mullen W., Nemzer B., Ou B., Stalmach A., Hunter J., Clifford M.N., Combet E., The antioxidant and chlorogenic acid profiles of whole coffee fruits are influenced by the extraction procedures, Journal of agricultural and food chemistry., 59(8) (2011) 3754-3762.
  • [18] Tan Z., Wang C., Yi Y., Wang H., Li M., Zhou W., Li F., Extraction and purification of chlorogenic acid from ramie (Boehmeria nivea L. Gaud) leaf using an ethanol/salt aqueous two-phase system, Separation and Purification Technology., 132 (2014) 396-400.
  • [19] Płotka J., Tobiszewski M., Sulej A.M., Kupska M., Górecki T., Namieśnik J., Green chromatography, Journal of Chromatography A., 1307 (2013) 1-20.
  • [20] Demirkaya-Miloglu F., Yaman M. E., Kadioglu Y., Optimization of a spectrofluorimetric method based on a central composite design for the determination of potassium losartan in pharmaceutical products, Brazilian Journal of Pharmaceutical Sciences., 50 (2014) 611-619.
  • [21] Theoret C.L., Update on wound repair, Clinical Techniques in Equine Practice., 3(2) (2004) 110-122.
  • [22] Zanca A., Flegg J.A., Osborne J. M., Push or Pull? Cell Proliferation and Migration During Wound Healing, Frontiers in Systems Biology., 2 (2022) 876075.
  • [23] Ayla Ş., Günal M.Y., Sayın Şakul A.A., Biçeroğlu Ö., Özdemir E.M., Okur M.E., Bilgiç B. E., Effects of Prunus spinosa L. fruits on experimental wound healing, Medeniyet medical journal., 32(3) (2017).
  • [24] Bigović D., Šavikin K., Janković T., Menković N., Zdunić G., Stanojković T., Djurić Z., Antiradical and cytotoxic activity of different Helichrysum plicatum flower extracts, Natural product communications., 6(6) (2011) 1934578X1100600617.
  • [25] Bigović D.J., Stević T.R., Janković T.R., Noveski N.B., Radanović D.S., Pljevljakušić D.S.,Djurić Z.R., Antimicrobial activity of Helichrysum plicatum DC, HEMIJSKA INDUSTRIJA (Chemical Industry)., 71(4) (2017) 337-342.
  • [26] Steenkamp V., Mathivha E., Gouws M.C., Van Rensburg C.E.J., Studies on antibacterial, antioxidant and fibroblast growth stimulation of wound healing remedies from South Africa, Journal of ethnopharmacology., 95(2-3) (2004) 353-357.
  • [27] Erel O., A new automated colorimetric method for measuring total oxidant status, Clinical biochemistry, 38(12) (2005) 1103-1111.
  • [28] Pereira J.C., Paulino C.L.D.A., Granja B.D.S., Santana A.E.G., Endres L.,Souza R.C.D., Allelopathic potential and identification of secondary metabolites in extracts of Canavalia ensiformis L, Revista Ceres., 65 (2018)243-252.
  • [29] Santana-Gálvez J., Cisneros-Zevallos L., Jacobo-Velázquez D.A.,Chlorogenic acid: Recent advances on its dual role as a food additive and a nutraceutical against metabolic syndrome, Molecules., 22(3) (2017) 358.
  • [30] Rofouei M.K., Kojoori S.M.H.,Moazeni-Pourasil R.S., Optimization of chlorogenic acid extraction from Elm tree, Ulmus minor Mill., fruits, using response surface methodology,Separation and Purification Technology., 256 (2021) 117773.
  • [31] Skoog D.A., Holler F.J., Crouch S.R., Principles of instrumental analysis, Cengage learning (2017).
  • [32] Gouveia S.C., Castilho P.C., Validation of a HPLC-DAD–ESI/MSn method for caffeoylquinic acids separation, quantification and identification in medicinal Helichrysum species from Macaronesia, Food research international., 45(1) (2012) 362-368.
  • [33] Silva L., Rodrigues A.M., Ciriani M., Falé P.L.V., Teixeira V., Madeira P.,Serralheiro M.L.M., Antiacetylcholinesterase activity and docking studies with chlorogenic acid, cynarin and arzanol from Helichrysum stoechas (Lamiaceae), Medicinal Chemistry Research., 26 (2017) 2942-2950.
  • [34] Dinç, E., Kemometri Çok Değişkenli Kalibrasyon Yöntemleri,Hacettepe Üniversitesi, Eczacılık Fakültesi Dergisi., (2007) 61-92.
  • [35] Özkorkmaz E.G., Özay Y., Yara iyileşmesi ve yara iyileşmesinde kullanılan bazı bitkiler, Türk Bilimsel Derlemeler Dergisi., (2) (2009) 63-67.
  • [36] Gundogdu G., Nalci K.A., Ugur Kaplan A.B., Gundogdu K., Demirci T., Demirkaya-Miloglu F., Cetin M.,The evaluation of the effects of nanoemulsion formulations containing boron and/or zinc on the wound healing in diabetic rats, The International Journal of Lower Extremity Wounds., 21(4) (2022) 492-501.
  • [37] Villegas J., McPhaul M., Establishment and culture of human skin fibroblasts, Current protocols in molecular biology., 71(1), (2005) 28-3.
  • [38] Moghadam S.E., Ebrahimi S. N., Salehi P., Moridi Farimani M., Hamburger M., Jabbarzadeh E., Wound healing potential of chlorogenic acid and Myricetin-3-O-β-Rhamnoside isolated from Parrotia persica, Molecules., 22(9) (2017) 1501.
  • [39] Alexandru V., Gaspar A., Savin S., Toma A., Tatia R., Gille E.,Phenolic content, antioxidant activity and effect on collagen synthesis of a traditional wound healing polyherbal formula, Studia Universitatis" Vasile Goldis" Arad. Seria Stiintele Vietii (Life Sciences Series)., 25(1) (2015) 41.
  • [40] Muniandy K., Gothai S., Tan W.S., Kumar S.S., Mohd Esa N., Chandramohan G., Arulselvan P., In vitro wound healing potential of stem extract of Alternanthera sessilis, Evidence-based complementary and alternative medicine., (2018) 2018.
  • [41] Gundogdu G., Gundogdu K., Nalci K. A., Demirkaya A. K., Yılmaz Tascı S., Demirkaya Miloglu F., Hacimuftuoglu, A., The effect of parietin isolated from rheum ribes l on in vitro wound model using human dermal fibroblast cells, The International Journal of Lower Extremity Wounds., 18(1) (2019) 56-64.

Year 2024, , 299 - 308, 30.06.2024

Abdulbaki Akpınar , Fatma Demirkaya Miloğlu , Gülşah Gündoğdu , Leyla Güven , Burak Bayrak , Yücel Kadıoğlu

https://doi.org/10.17776/csj.1440382

Abstract

References

  • [1] Kremr D., Bajer T., Bajerová P., Surmová S., Ventura K., Unremitting problems with chlorogenic acid nomenclature: A
  • [2] Review, Química Nova., 39 (2016) 530-533.
  • [3] Naveed M., Hejazi V., Abbas M., Kamboh A.A., Khan G.J., Shumzaid M., XiaoHui Z., Chlorogenic acid (CGA): A pharmacological review and call for further research, Biomedicine & pharmacotherapy., 97 (2018) 67-74.
  • [4] Budama-Kılınç Y., Klorojenik Asit Yüklü PLGA Nanopartiküllerinin Üretimi ve Antimikrobiyal Etkinliğinin Belirlenmesi, Türk Mikrobiyoloji Cemiyeti Dergisi., 49(1) (2019) 47-54.
  • [5] Feng R., Lu Y., Bowman L.L., Qian,Y., Castranova V., Ding M., Inhibition of activator protein-1, NF-κB, and MAPKs and induction of phase 2 detoxifying enzyme activity by chlorogenic acid, Journal of Biological Chemistry., 280 (2005) 27888-27895.
  • [6] Zhong C., Wall N.R., Zu Y., Sui G., Therapeutic application of natural medicine monomers in cancer treatment, Current Medicinal Chemistry., 24(34) (2017) 3681-3697.
  • [7] Bagdas D., Gul N.Y., Topal A., Tas S., Ozyigit M.O., Cinkilic N., Gurun M.S., Pharmacologic overview of systemic chlorogenic acid therapy on experimental wound healing, Naunyn-Schmiedeberg's archives of pharmacology., 387 (2014) 1101-1116.
  • [8] Chen W.C., Liou S.S., Tzeng T.F., Lee S.L., Liu I.M., Effect of topical application of chlorogenic acid on excision wound healing in rats, Planta medica., (2013) 616-621.
  • [9] Chaowuttikul C., Palanuvej C., Ruangrungsi N., Quantification of chlorogenic acid, rosmarinic acid, and caffeic acid contents in selected Thai medicinal plants using RP-HPLC-DAD, Brazilian Journal of Pharmaceutical Sciences., 56 (2020) e17547.
  • [10] Kolaylı S., Şahin H., Ulusoy E., Tarhan Ö., Phenolic composition and antioxidant capacities of Helichrysum plicatum, Hacettepe Journal of Biology and Chemistry., 38(4) (2010) 269-276.
  • [11] Albayrak S., Aksoy A., Sagdic O., Hamzaoglu E.,Compositions, antioxidant and antimicrobial activities of Helichrysum (Asteraceae) species collected from Turkey, Food chemistry., 119(1) (2010) 114-122.
  • [12] Albayrak S., Aksoy A., Sağdıç O., Budak Ü., Phenolic compounds and antioxidant and antimicrobial properties of Helichrysum species collected from eastern Anatolia, Turkey,Turkish Journal of Biology., 34(4) (2010) 463-473.
  • [13] Ayelign A., Sabally K., Determination of chlorogenic acids (CGA) in coffee beans using HPLC, American Journal of Research Communication., 1(2) (2013) 78-91.
  • [14] De Luca S., Ciotoli E., Biancolillo A., Bucci R., Magrì A.D.,Marini F., Simultaneous quantification of caffeine and chlorogenic acid in coffee green beans and varietal classification of the samples by HPLC-DAD coupled with chemometrics, Environmental Science and Pollution Research., 25 (2018) 28748-28759.
  • [15] Li H., Chen B., Yao S., Application of ultrasonic technique for extracting chlorogenic acid from Eucommia ulmodies Oliv.(E. ulmodies), Ultrasonics sonochemistry., 12(4) (2005) 295-300.
  • [16] Zhang B., Yang R., Liu C.Z., Microwave-assisted extraction of chlorogenic acid from flower buds of Lonicera japonica Thunb, Separation and Purification Technology., 62(2) (2008) 480-483.
  • [17] Mullen W., Nemzer B., Ou B., Stalmach A., Hunter J., Clifford M.N., Combet E., The antioxidant and chlorogenic acid profiles of whole coffee fruits are influenced by the extraction procedures, Journal of agricultural and food chemistry., 59(8) (2011) 3754-3762.
  • [18] Tan Z., Wang C., Yi Y., Wang H., Li M., Zhou W., Li F., Extraction and purification of chlorogenic acid from ramie (Boehmeria nivea L. Gaud) leaf using an ethanol/salt aqueous two-phase system, Separation and Purification Technology., 132 (2014) 396-400.
  • [19] Płotka J., Tobiszewski M., Sulej A.M., Kupska M., Górecki T., Namieśnik J., Green chromatography, Journal of Chromatography A., 1307 (2013) 1-20.
  • [20] Demirkaya-Miloglu F., Yaman M. E., Kadioglu Y., Optimization of a spectrofluorimetric method based on a central composite design for the determination of potassium losartan in pharmaceutical products, Brazilian Journal of Pharmaceutical Sciences., 50 (2014) 611-619.
  • [21] Theoret C.L., Update on wound repair, Clinical Techniques in Equine Practice., 3(2) (2004) 110-122.
  • [22] Zanca A., Flegg J.A., Osborne J. M., Push or Pull? Cell Proliferation and Migration During Wound Healing, Frontiers in Systems Biology., 2 (2022) 876075.
  • [23] Ayla Ş., Günal M.Y., Sayın Şakul A.A., Biçeroğlu Ö., Özdemir E.M., Okur M.E., Bilgiç B. E., Effects of Prunus spinosa L. fruits on experimental wound healing, Medeniyet medical journal., 32(3) (2017).
  • [24] Bigović D., Šavikin K., Janković T., Menković N., Zdunić G., Stanojković T., Djurić Z., Antiradical and cytotoxic activity of different Helichrysum plicatum flower extracts, Natural product communications., 6(6) (2011) 1934578X1100600617.
  • [25] Bigović D.J., Stević T.R., Janković T.R., Noveski N.B., Radanović D.S., Pljevljakušić D.S.,Djurić Z.R., Antimicrobial activity of Helichrysum plicatum DC, HEMIJSKA INDUSTRIJA (Chemical Industry)., 71(4) (2017) 337-342.
  • [26] Steenkamp V., Mathivha E., Gouws M.C., Van Rensburg C.E.J., Studies on antibacterial, antioxidant and fibroblast growth stimulation of wound healing remedies from South Africa, Journal of ethnopharmacology., 95(2-3) (2004) 353-357.
  • [27] Erel O., A new automated colorimetric method for measuring total oxidant status, Clinical biochemistry, 38(12) (2005) 1103-1111.
  • [28] Pereira J.C., Paulino C.L.D.A., Granja B.D.S., Santana A.E.G., Endres L.,Souza R.C.D., Allelopathic potential and identification of secondary metabolites in extracts of Canavalia ensiformis L, Revista Ceres., 65 (2018)243-252.
  • [29] Santana-Gálvez J., Cisneros-Zevallos L., Jacobo-Velázquez D.A.,Chlorogenic acid: Recent advances on its dual role as a food additive and a nutraceutical against metabolic syndrome, Molecules., 22(3) (2017) 358.
  • [30] Rofouei M.K., Kojoori S.M.H.,Moazeni-Pourasil R.S., Optimization of chlorogenic acid extraction from Elm tree, Ulmus minor Mill., fruits, using response surface methodology,Separation and Purification Technology., 256 (2021) 117773.
  • [31] Skoog D.A., Holler F.J., Crouch S.R., Principles of instrumental analysis, Cengage learning (2017).
  • [32] Gouveia S.C., Castilho P.C., Validation of a HPLC-DAD–ESI/MSn method for caffeoylquinic acids separation, quantification and identification in medicinal Helichrysum species from Macaronesia, Food research international., 45(1) (2012) 362-368.
  • [33] Silva L., Rodrigues A.M., Ciriani M., Falé P.L.V., Teixeira V., Madeira P.,Serralheiro M.L.M., Antiacetylcholinesterase activity and docking studies with chlorogenic acid, cynarin and arzanol from Helichrysum stoechas (Lamiaceae), Medicinal Chemistry Research., 26 (2017) 2942-2950.
  • [34] Dinç, E., Kemometri Çok Değişkenli Kalibrasyon Yöntemleri,Hacettepe Üniversitesi, Eczacılık Fakültesi Dergisi., (2007) 61-92.
  • [35] Özkorkmaz E.G., Özay Y., Yara iyileşmesi ve yara iyileşmesinde kullanılan bazı bitkiler, Türk Bilimsel Derlemeler Dergisi., (2) (2009) 63-67.
  • [36] Gundogdu G., Nalci K.A., Ugur Kaplan A.B., Gundogdu K., Demirci T., Demirkaya-Miloglu F., Cetin M.,The evaluation of the effects of nanoemulsion formulations containing boron and/or zinc on the wound healing in diabetic rats, The International Journal of Lower Extremity Wounds., 21(4) (2022) 492-501.
  • [37] Villegas J., McPhaul M., Establishment and culture of human skin fibroblasts, Current protocols in molecular biology., 71(1), (2005) 28-3.
  • [38] Moghadam S.E., Ebrahimi S. N., Salehi P., Moridi Farimani M., Hamburger M., Jabbarzadeh E., Wound healing potential of chlorogenic acid and Myricetin-3-O-β-Rhamnoside isolated from Parrotia persica, Molecules., 22(9) (2017) 1501.
  • [39] Alexandru V., Gaspar A., Savin S., Toma A., Tatia R., Gille E.,Phenolic content, antioxidant activity and effect on collagen synthesis of a traditional wound healing polyherbal formula, Studia Universitatis" Vasile Goldis" Arad. Seria Stiintele Vietii (Life Sciences Series)., 25(1) (2015) 41.
  • [40] Muniandy K., Gothai S., Tan W.S., Kumar S.S., Mohd Esa N., Chandramohan G., Arulselvan P., In vitro wound healing potential of stem extract of Alternanthera sessilis, Evidence-based complementary and alternative medicine., (2018) 2018.
  • [41] Gundogdu G., Gundogdu K., Nalci K. A., Demirkaya A. K., Yılmaz Tascı S., Demirkaya Miloglu F., Hacimuftuoglu, A., The effect of parietin isolated from rheum ribes l on in vitro wound model using human dermal fibroblast cells, The International Journal of Lower Extremity Wounds., 18(1) (2019) 56-64.
There are 41 citations in total.

Details

Primary Language English
Subjects Quality Assurance, Chemometrics, Traceability and Metrological Chemistry
Journal Section Natural Sciences
Authors

Abdulbaki Akpınar 0000-0001-7318-6529

Fatma Demirkaya Miloğlu 0000-0001-5729-7181

Gülşah Gündoğdu 0000-0002-9924-5176

Leyla Güven 0000-0002-3189-6415

Burak Bayrak 0000-0001-6550-6916

Yücel Kadıoğlu 0000-0001-6590-7306

Publication Date June 30, 2024
Submission Date February 20, 2024
Acceptance Date June 21, 2024
Published in Issue Year 2024

Cite

APA Akpınar, A., Demirkaya Miloğlu, F., Gündoğdu, G., Güven, L., et al. (2024). Chlorogenic Acid: HPLC Quantification and In Vitro Assessment of Proliferative and Migration Effects on Human Dermal Fibroblast Cells. Cumhuriyet Science Journal, 45(2), 299-308. https://doi.org/10.17776/csj.1440382