HPTLC-DPPH• and HPTLC-tyrosinase methods for hot water-soluble contents of kumquat, limequat and Mexican lime fruit powders

Hale Gamze Ağalar; Burak Temiz

HPTLC-DPPH• and HPTLC-tyrosinase methods for hot water-soluble contents of kumquat, limequat and Mexican lime fruit powders

Abstract

In the present study, small Citrus fruits which are cultivated in our country and have an economic value were chosen as research materials. It was aimed to do fingerprint analysis of the fruits by HPTLC method. DPPH• scavenging and antityrosinase potentials were determined in vitro. Extracts showed noticable activities, but these effects were not comparable with positive controls. HPTLC-bioautographic assays and Mass Spectrometry analysis were employed to identify bioactive compound in the extracts. Phloretin-di-C-glucoside was tentatively identified as DPPH• scavenging and antityrosinase active compound in kumquat and limequat extracts.

Keywords

References

[1] Butelli E, Garcia-Lor A, Licciardello C, Las Casas G, Hill L, Recupero GR, Keremane ML, Ramadugu C, Krueger R, Xu Q, Deng X, Fanciullino AL, Froelicher Y, Navarro L, Martin C. Changes in anthocyanin production during domestication of Citrus. Plant Physiol. 2017; 173(4): 2225-2242.
[2] Janick J. The origins of fruits, fruit growing, and fruit breeding. Plant Breed Rev. 2005; 25: 255-320.
[3] Abobatta WF. Nutritional benefits of Citrus fruits. Am J Biomed Sci Res. 2019; 3(4): 303-306.
[4] Brito A, Ramirez JE, Areche C, Sepulveda B, Simirgiotis MJ. HPLC-UV-MS profiles of phenolic compounds and antioxidant activity of fruits from three citrus species consumed in Northern Chile. Molecules. 2014; 19: 17400-17421.
[5] Zhao Y, Kao CP, Liao CR, Wu KC, Zhou X, Ho YL, Chang YS. Chemical compositions, chromatographic fingerprints and antioxidant activities of Citri exocarpium rubrum (Juhong). Chin Med. 2017; 12: 6.
[6] Cheng LP, Ren YJ, Lin DB, Peng S, Zhong B, Ma ZC. The anti-inflammatory properties of Citrus wilsonii Tanaka extract in LPS-induced RAW 264.7 and primary mouse bone marrow-derived dendritic cells. Molecules. 2017; 22(7): 1213.
[7] Mandalari G, Bisignano C, Cirmi S, Navarra N. Effectiveness of Citrus fruits on Helicobacter pylori. Evid Based Complement Altern Med. 2017; 2017: 8379262.
[8] Onuma W, Asai D, Tomono S, Miyamoto S, Fujii G, Hamoya T, Nagano A, Takahashi S, Masumori S, Miyoshi N, Wakabayashi K, Mutoh M. Anticarcinogenic effects of dried Citrus peel in colon carcinogenesis due to inhibition of oxidative stress. Nutr Cancer Int J. 2017; 69(6): 855-861.

[9] Cirmi S, Maugeri A, Ferlazzo N, Gangemi S, Calapai G, Schumacher U, Navarra M. Anticancer potential of citrus juices and their extracts: A systematic review of both preclinical and clinical studies. Front Pharmacol. 2017; 8: 420.
[10] Kim AH, Kim HJ, Ryu R, Han HJ, Han YJ, Lee MK, Choi MS, Park YB. A mixture of ethanol extracts of Persimmon leaf and Citrus junos Sieb improves blood coagulation parameters and ameliorates lipid metabolism disturbances caused by diet-induced obesity in C57BL/6J mice. J Microbiol Biotechnol. 2016; 26(2): 295-308.
[11] Curro M, Risitano R, Ferlazzo N, Cirmi S, Gangemi C, Caccamo D, Ientile R, Navarra M. Citrus bergamia juice extract attenuates beta-amyloid-induced pro-inflammatory activation of THP-1 cells through MAPK and AP-1 pathways. Sci Rep. 2016; 6: 20809.
[12] Sadek ES, Makris DP, Kefalas P. Polyphenolic composition and antioxidant characteristics of kumquat (Fortunella margarita) peel fractions. Plant Foods Hum Nutr. 2009; 64: 297-302.
[13] Olcay N, Demir MK. Nutritional composition and functional properties of Kumquat (Fortunella spp.). J Inst Sci Technol. 2019; 9(4): 2124-2132.
[14] Agalar HG, Temiz B, Demirci B, Baser KHC. Drying effects on the volatile compounds of kumquat, limequat and Mexican lime fruits. J Essent Bear Plants. 2020; 23(6): 1395-1408.
[15] Casilli A, Decorzant E, Jaquier A, Delort E. Multidimensional gas chromatography hyphenated to mass spectrometry and olfactometry for the volatile analysis of citrus hybrid peel extract. J Chrom A. 2014; 1373: 169-178.
[16] Ağalar HG, Temiz B, Demirci B, Başer KHC. Direct SPME/GC-MS analyzes of small citrus fruits cultivated in Turkey. EMU J Pharm Sci. 2020; 3(3): 140-152.
[17] Viloria Z, Grosser JW, Bracho B. Immature embryo rescue, culture and seedling development of acid citrus fruit derived from interploid hybridization. Plant Cell Tiss Organ Cult. 2005; 82(2): 159-167.
[18] Barreca D, Bellocco E, Caristi C, Leuzzi U, Gattuso G. Kumquat (Fortunella japonica Swingle) juice: flavonoid distribution and antioxidant properties. Food Res Int. 2011; 44(7): 2190-2197.
[19] Lou SN, Lai YC, Huang JD, Ho CT, Ferng LHA, Chang YC. Drying effect on flavonoid composition and antioxidant activity of immature kumquat. Food Chem. 2015; 171: 356-363.
[20] Tan S, Zhao XJ, Yang Y, Ke ZL, Zhou ZQ. Chemical profiling using UPLC Q-TOF/MS and antioxidant activities of Fortunella fruits. J Food Sci. 2016; 81(79): C1646-C1653.
[21] Goh RMV, Pua A, Liu SQ, Lassabliere B, Leong KC, Sun J, Tan LP, Yu B. Characterisation of volatile compounds in kumquat and calamansi peel oil extracts. J Essent Oil Bear Plants. 2020; 23(5): 953-969.
[22] Nouri A, Shafaghatlonbar A. Chemical constituents and antioxidant activity of essential oil and organic extract from the peel and kernel parts of Citrus japonica Thunb. (kumquat) from Iran. Nat Prod Res. 2016; 30(9): 1093-1097.
[23] Lin CC, Hung PF, Ho SC. Heat treatment enhances the NO-suppressing and peroxynitrite-intercepting activities of kumquat (Fortunella margarita Swingle) peel. Food Chem. 2008; 109(1): 95-103.
[24] Ramful D, Tarnus E, Aruoma OI, Bourdon E, Bahorun T. Polyphenol composition, vitamin C content and antioxidant capacity of Mauritian citrus fruit pulps. Food Res Int. 2011; 44(7): 2088-2099.
[25] Damian-Reyna AA, Gonzalez-Hernandez JC, Maya-Yescas R, Cortes-Penagos CD, Chavez-Parga MD. Polyphenolic content and bactericidal effect of Mexican Citrus limetta and Citrus reticulata. J Food Sci Tech. 2017; 54(2): 531-537.
[26] Craske JD, Suryadi N, Wootton M. A comparison of the peel oil components of Australian native lime (Microcitrus australe) and Mexican lime (Citrus aurantifolia Swingle). J Sci Food Agric. 2005; 85(3): 522-525.
[27] Ramful D, Bahorun T, Bourdon E, Tarnus E, Aruoma OI. Bioactive phenolics and antioxidant propensity of flavedo extracts of Mauritian citrus fruits: Potential prophylactic ingredients for functional foods application. Toxicol. 2010; 278(1): 75-87.
[28] Settanni L, Palazzolo E, Guarrasi V, Aleo A, Mammina C, Moschetti G, Germana MA. Inhibition of foodborne pathogen bacteria by essential oils extracted from citrus fruits cultivated in Sicily. Food Control. 2012; 26(2): 326-330.
[29] Spadaro F, Costa R, Circosta C, Occhiuto F. Volatile composition and biological activity of key lime Citrus aurantifolia essential oil. Nat Prod Commun. 2012; 7(11): 1523-1526.
[30] Aldana DS, Andrade-Ochoa S, Aguilar CN, Contreras-Esquivel JC, Nevarez-Moorillon GV. Antibacterial activity of pectic-based edible films incorporated with Mexican lime essential oil. Food Control. 2015; 50: 907-912.
[31] Dominguez-Vigil IG, Camacho-Corona MD, Heredia-Rojas JA, Vargas-Villarreal J, Rodriguez-De la Fuente AO, Heredia-Rodriguez O, Mata-Cardenas BD. Anti-giardia activity of hexane extract of Citrus aurantifolia (Christim) Swingle and some of its constituents. Afr J Tradit Complement Altern Med. 2015; 12(2): 55-59.
[32] Patil JR, Murthy KNC, Jayapraskasha GK, Chetti MB, Patil BS. Bioactive compounds from Mexican lime (Citrus aurantifolia) juice induce apoptosis in human pancreatic cells. J Agric Food Chem. 2009; 57(22): 10933-10942.
[33] Orozco-Santos M, Robles-Gonzalez M, Hernandez-Fuentes LM, Velazquez-Monreal JJ, Bermudez-Guzman MD, Manzanilla-Ramirez M, Manzo-Sanchez G, Nieto-Angel D. Use of oils and plant extracts to control Diaphorina citri kuwayama1 on Mexican Lime in the dry tropic of Mexico. Southwest Entomol. 2016; 41(4): 1051-1066.
[34] Lou SN, Lai YC, Huang JD, Ho CT, Ferng LHA, Chang YC. Drying effect on flavonoid composition and antioxidant activity of immature kumquat. Food Chem. 2015; 171: 356-363.
[35] Singleton VL, Orthofer R, Lamuela-Raventos RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methodol Enzymol. 1999; 299: 152-178.
[36] Duymuş HG, Göger F, Başer KHC. In vitro antioxidant properties and anthocyanin compositions of elderberry extracts. Food Chem. 2014; 155: 112-119.
[37] Likhitwitayawuid K, Sritularak B. A new dimeric stilbene with tyrosinase inhibitiory activity from Artocarpus gomezianus. J Nat Prod. 2001; 64(11): 1457-1459.
[38] Orcini F, Vovk I, Glavnik V, Jug U, Corradini D. HPTLC, HPTLC-MS/MS and HPTLC methods for analyses of flavonoids and their antioxidant activity in Cyclanthera pedate leaves, fruits and dietary supplement. J Liquid Chrom Relat Tech. 2019; 42: 290-301.
[39] Revoltella S, Rainer B, Waltenberger B, Pagitz K, Schwaiger S, Stuppner H. HPTLC autography based screening and isolation of mushroom tyrosinase inhibitors of European plant species. Chem Biodivers. 2019; 16(3): e1800541.
[40] Zolghadri S, Bahrami A, Khan MTH, Munoz-Munoz J, Garcia-Molina F, Garcia-Canovas F, Saboury AA. A compherensive review on tyrosinase inhibitors. J Ezyme Inhib Med Chem. 2019; 34(1): 279-309.
[41] Farras A, Casedas G, Les F, Terrado EM, Mitjans M, Lopez V. Evaluation of antityrosinase and antioxidant properties of four fern species for potential cosmetic applications. Forests. 2019; 10(2): 179.
[42] Wang HM, Chen CY, Chen CY, Ho ML, Chou YT, Chang HC, Lee CH, Wang CZ, Chu IM. (-)-N-Formylanonaine from Michelia alba as a human tyrosinase inhibitor and antioxidant. Bioorg Med Chem. 2010; 18(14): 5241-5247.
[43] Batubara I, Darusman LK, Mitsunaga T, Rahminiwati M, Djauhari E. Potency of Indonesian medicinal plants as tyrosinase inhibitor and antioxidant agent. J Biol Sci. 2010; 10(2), 138-144.
[44] Ogawa K, Kawasaki A, Omura M, Yoshida T, Ikoma Y, Yano M. 3’,5’-D-C-β-glucopyranosylphloretin, a flavonoid characteristic of the genus Fortunella. Phytochem. 2001; 57(5): 737-742.
[45] Lou SN, Lai YC, Hsu YS, Ho CT. Phenolic content, antioxidant activity and effective compounds of kumquat extracted by different solvents. Food Chem. 2016; 197: 1-6.
[46] Nithiya T, Udayakumar R. In vitro antioxidant properties of phloretin—An important phytocompound. J Biosci Med. 2016; 4(1): 85-90.
[47] Zhang T, Liang J, Wang P, Xu Y, Wang Y, Wei X, Fan M. Purification and characterization of a novel phloretin-2′-O- glycosyltransferase favoring phloridzin biosynthesis. Sci Rep. 2016; 6: 35274.
[48] Lou SN, Yu MW, Ho CT. Tyrosinase inhibitory components of immature calamondin peel. Food Chem. 2012; 135(3): 1091-1096.
[49] Beelders T, De Beer D, Stander MA, Joubert E. Comprehensive phenolic profiling of Cyclopia genistoides (L.) Vent. by LC-DAD-MS and-MS/MS reveals novel xanthone and benzophenone constituents. Molecules. 2014; 19(8):11760- 11790.
[50] De Beer D, Schulze AE, Joubert E, De Villiers A, Malherbe CJ, Stander MA. Food ingredient extracts of Cyclopia subternata (Honeybush): variation in phenolic composition and antioxidant capacity. Molecules. 2012; 17(12): 14602- 14624.
[51] Schulze AE, Beelders T, Koch IS, Erasmus LM, De Beer D, Joubert E. Honeybush herbal teas (Cyclopia spp.) contribute to high levels of dietary exposure to xanthones, benzophenones, dihydrochalcones and other bioactive phenolics. J Food Compos Anal. 2015; 44: 139-148.
[52]Barreca D, Bellocco E, Lagana G, Ginestra G, Bisignano C. Biochemical and antimicrobial activity of phloretin and its glycosilated derivatives present in apple and kumquat. Food Chem. 2014; 160: 292-297.

Details

Primary Language

English

Subjects

Pharmacognosy

Journal Section

Research Article

Authors

Hale Gamze Ağalar ^*
Türkiye

Burak Temiz
Türkiye

Publication Date

June 28, 2025

Submission Date

February 15, 2021

Acceptance Date

July 5, 2021

Published in Issue

Year 2021 Volume: 25 Number: 5

IZ

https://izlik.org/JA92GW84MN

Cite

RIS / Bibtex

APA

Ağalar, H. G., & Temiz, B. (2025). HPTLC-DPPH• and HPTLC-tyrosinase methods for hot water-soluble contents of kumquat, limequat and Mexican lime fruit powders. Journal of Research in Pharmacy, 25(5), 569-580. https://izlik.org/JA92GW84MN

AMA

1.Ağalar HG, Temiz B. HPTLC-DPPH• and HPTLC-tyrosinase methods for hot water-soluble contents of kumquat, limequat and Mexican lime fruit powders. J. Res. Pharm. 2025;25(5):569-580. https://izlik.org/JA92GW84MN

Chicago

Ağalar, Hale Gamze, and Burak Temiz. 2025. “HPTLC-DPPH• and HPTLC-Tyrosinase Methods for Hot Water-Soluble Contents of Kumquat, Limequat and Mexican Lime Fruit Powders”. Journal of Research in Pharmacy 25 (5): 569-80. https://izlik.org/JA92GW84MN.

EndNote

Ağalar HG, Temiz B (July 1, 2025) HPTLC-DPPH• and HPTLC-tyrosinase methods for hot water-soluble contents of kumquat, limequat and Mexican lime fruit powders. Journal of Research in Pharmacy 25 5 569–580.

IEEE

[1]H. G. Ağalar and B. Temiz, “HPTLC-DPPH• and HPTLC-tyrosinase methods for hot water-soluble contents of kumquat, limequat and Mexican lime fruit powders”, J. Res. Pharm., vol. 25, no. 5, pp. 569–580, July 2025, [Online]. Available: https://izlik.org/JA92GW84MN

ISNAD

Ağalar, Hale Gamze - Temiz, Burak. “HPTLC-DPPH• and HPTLC-Tyrosinase Methods for Hot Water-Soluble Contents of Kumquat, Limequat and Mexican Lime Fruit Powders”. Journal of Research in Pharmacy 25/5 (July 1, 2025): 569-580. https://izlik.org/JA92GW84MN.

JAMA

1.Ağalar HG, Temiz B. HPTLC-DPPH• and HPTLC-tyrosinase methods for hot water-soluble contents of kumquat, limequat and Mexican lime fruit powders. J. Res. Pharm. 2025;25:569–580.

MLA

Ağalar, Hale Gamze, and Burak Temiz. “HPTLC-DPPH• and HPTLC-Tyrosinase Methods for Hot Water-Soluble Contents of Kumquat, Limequat and Mexican Lime Fruit Powders”. Journal of Research in Pharmacy, vol. 25, no. 5, July 2025, pp. 569-80, https://izlik.org/JA92GW84MN.

Vancouver

1.Hale Gamze Ağalar, Burak Temiz. HPTLC-DPPH• and HPTLC-tyrosinase methods for hot water-soluble contents of kumquat, limequat and Mexican lime fruit powders. J. Res. Pharm. [Internet]. 2025 Jul. 1;25(5):569-80. Available from: https://izlik.org/JA92GW84MN