Clinopodium vulgare L. subsp. vulgare Ekstresinin Antioksidan, Antimikrobial, Tirozinaz İnhibitor Aktiviteleri ve RP-HPLC ile Fenolik Bileşiklerinin Araştırılması

Yıl 2018, , 230 - 238, 31.07.2018

Rezzan Aliyazıcıoğlu , Sıla Özlem Şener , Nuriye Korkmaz , Şeyda Akkaya Ufuk Özgen Şengül Alpay Karaoğlu

https://doi.org/10.17714/gumusfenbil.306925

Öz

Clinopodium vulgare L. subsp. vulgare, Clinopodium
cinsine ve Lamiaceae familyasına mensup çok yıllık aromatik otsu bir bitkidir.
Bu cins bitkiler sıklıkla geleneksel tedavide kullanılmaktadır. Farklı
Clinopodium türlerinin farklı ekstrelerinin antioksidan, antimikrobiyal,
tirozinaz inhibitör etkilerini ve HPLC ile fenolik bileşenlerinin analizini
konu alan çalışmalar olmasına rağmen, Clinopodium vulgare L. subsp. vulgare ekstresinin
bu özelliklerinin tümünü içeren herhangi bir çalışma bulunmamaktadır. Bu
çalışmanın amacı C. vulgare  subsp.
vulgare metanol ekstresinin antioksidan, antimikrobiyal, tirozinaz inhibitör
aktivitelerini ve HPLC analizi ile fenolik madde içeriğini belirlemektir. Ekstrenin fenolik
bileşenleri ters faz yüksek performanslı sıvı kromatografisi (RP-HPLC) ile
belirlendi. Antioksidan, tirozinaz inhibitör aktivite spektrofotometrik
yöntemlerle ve antimikrobiyal aktivite disk difüzyon metodu ile incelendi. Ekstrenin toplam fenolik madde miktarı 27.9±0.4 mg gallik
asit eşdeğeri/g numune, 2,2-difenil-1-pikrilhidrazil (DPPH) radikal süpürme kapasitesi
IC50 değeri 0.114±0.0004 mg/mL ve ferrik indirgeyici antioksidan güç
(FRAP) değeri 1556±3 µM troloks eşdeğeri/g numune olarak hesaplandı. Tirozinaz
inhibisyon çalışma sonucuna göre ekstrenin IC50 değeri kojik asit
standardından yüksek bulundu. HPLC ile analiz sonucunda protokatekuik asit,
klorojenik asit, vanilin, sinapik asit ve benzoik asit tespit edildi. Ekstre,
asid-hızlı bakteri (M. smegmatis), bazı gram pozitif (S. aureus ve B. cereus)
ve bazı gram negatif (Y. pseudotuberculosis) bakterilere karşı ılımlı
antibakteriyal aktivite gösterdi. Ancak C. albicans and S. cerevisiae türlerine
karşı hiçbir antifungal aktivite göstermedi. Bu çalışmanın sonuçlarına göre, C. vulgare subsp. vulgare yeni
farmasotiklerin geliştirilmesinde potansiyel bir kaynak olarak düşünülebilir. 

Anahtar Kelimeler

Antioksidan, Antimikrobiyal, Tirozinaz İnhibitör Aktivite, HPLC

Investigation of Phenolic Compounds by RP-HPLC and Antioxidant, Antimicrobial, Tyrosinase Inhibitor Activities of Clinopodium vulgare L. subsp. vulgare Extract

Öz

Clinopodium vulgare L. subsp. vulgare is a perennial
aromatic herbaceous plant belonging to Clinopodium genus and Lamiaceae family.
This genus plants are often used in traditional therapy. There are no studies
involving the whole of these properties of the Clinopodium vulgare L. subsp.
vulgare extract, although there are studies about antioxidant, antimicrobial,
tyrosinase inhibitor effects and analysis of phenolic compounds by HPLC of
different extracts of different Clinopodium genus. The purpose of this study
was to determine the phenolic composition by HPLC, and antioxidant,
antimicrobial, and tyrosinase inhibitor activity of methanolic extract of C. vulgare  subsp. vulgare. The phenolic compounds were determined by reverse
phase high performance liquid chromatography (RP-HPLC). The antioxidant, tyrosinase
inhibitor, and antimicrobial activities of the extract were examined by spectrophotometric
methods, and disc diffusion method, respectively. The total phenolic content
(TPC), 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity (DPPH), and
ferric reducing antioxidant power (FRAP) values of the extract were found
27.9±0.4 mg gallic acid equivalents per g sample, 0.114±0.0004 mg/mL, and 1556±3
µM trolox equivalents per g sample, respectively. IC50 value of the extract according to the
tyrosinase inhibitition study results was found higher than kojic acid standard.
The protocatechuic acid, chlorogenic acid, vanillin, sinapic acid, benzoic acid
were detected by HPLC analysis. The extract exhibited while moderate
antibacterial activity against an acid-fast bacterium (M. smegmatis), some gram
pozitive (S. aureus and B. cereus) and gram negative (Y. pseudotuberculosis)
bacteria. But, antifungal activity was not showed against C. albicans and S. cerevisiae species. According to
the results of this study, C. vulgare  subsp. vulgare can be
considered as a potential source for developing new pharmaceuticals.

Anahtar Kelimeler

Antioxidants, Antimicrobial, Tyrosinase inhibition, HPLC

Kaynakça

• Aliyazicioglu, R., Demir, S., Badem, M., Sener, S.O., Korkmaz, N., Demi, E.A., Ozgen, U., Karaoglu, S.A., Aliyazicioglu, Y., 2017. Antioxidant, Antigenotoxic, Antimicrobial Activities and Phytochemical Analysis of Dianthus carmelitarum, Records of Natural Products, 11, 3, 270-284.
• Asami, S., Manabe, H., Miyake, J., Tsurudome, Y., Hirano, T., Yamaguchi, R., Itoh, H. ve Kasai, H., 1997. Cigarette smoking induces an increase in oxidative DNA damage, 8-hydroxydeoxyguanosine, in a central site of the human lung, Carcinogenesis, 18, 9, 1763–1766.
• Benzie, I.F.F. ve Strain, J.J., 1996. The Ferric Reducing Ability of Plasma (FRAP) as a Measure of ‘‘Antioxidant Power’’: The FRAP Assay, Analytical Biochemistry, 239, 70–76.
• Birinci, S., 2008. Doğu Karadeniz Bölgesinde Doğal Olarak Bulunan Faydalı Bitkiler ve Kullanım Alanlarının Araştırılması. Yüksek Lisans Tezi, Çukurova Üniversitesi Fen Bilimleri Enstitüsü. Adana, 181s.
• Blois, M.S., 1958. Antioxidant determinations by the use of stable free radical, Nature, 181, 1199-1200.
• Cao, G. ve Prior, R.I., 1999. The Measurement of Oxygen Radical Absorbance Capacity in Biological Samples, Methods in Enzymology, 299, 50-62.
• Chan, E.W.C., Lim, Y.Y., Wong, L.F., Lianto, F.S., Wong, S.K., Lim, K.K., Joe, C.E. ve Lim, T.Y., 2008. Antioxidant and tyrosinase inhibition properties of leaves and rhizomes of ginger species, Food Chemistry, 109, 477–483.
• Chen, J.S., Wei, C. ve Marshall, M.R., 1991. Inhibition mechanism of kojic acid on polyphenol oxidase, Journal of Agricultural and Food Chemistry, 39:1897-1901.
• Çelikol, D., 2015. Hypericum perforatum bitkisinden elde edilen kantaron yağının yara iyileşmesi üzerine etkilerinin deneysel olarak incelenmesi. Doktora tezi, Cumhuriyet Üniversitesi, Ağız Diş ve Çene Cerrahisi Anabilim Dalı, Sivas, 196s.
• Dhalla, N.S., Temsah, R.M. ve Netticadan, T., 2000. Role of oxidative stress in cardiovascular diseases, Journal of Hypertension, 18, 655–673.
• Dzhambazov, B., Daskalova, S., Monteva, A., Popov, N., 2002. In vitro screening for antitumour activity of Clinopodium vulgare L. (Lamiaceae) extracts, Biological and Pharmaceutical Bulletin Journal, 25, 499–504.
• Eruygur, N., 2014. Türkiye’de yetişen bazı Echium türlerinin yara iyileştirici aktivitesinin araştırılması, Doktora tezi, Gazi Üniversitesi Farmakognozi Ana Bilim Dalı, Ankara, 176s.
• Gholamhoseinian A., ZohreRazmi Z., 2012. Screening the methanolic extracts of some plants for tyrosinase inhibitory activity. Toxicology Environment Chemistry, 94, 310-318.
• Halliwell, B., 2007. Dietary polyphenols: Good, bad or indifferent for your health? Cardiovascular Research, 73, 341-347.
• Kaur, C. and Kapoor, H.C., 2001. Antioxidants in fruits and vegetables-the millennium’s health, International Journal of Food Science & Technology, 36, 703-725.
• Kasparova, S., Brezova, V., Valko, M., Horecky, J., Mlynarik, V., Liptaj, T., Vancová, O., Ulicná, O. ve Dobrota, D., 2005. Study of the oxidative stress in a rat model of chronic brain hypoperfusion, Neurochemistry International, 46, 601–611.
• Kokdil, G., 1998. Composition of the essential oil of Clinopodium vulgare ssp. arundanum (Boiss.) from two different localities in Turkey, Flavour and Fragrance Journal, 13, 170–172.
• Lu, X., Wang, J., Al-Qadiri, H.M., Ross, C.F., Powers, J.R., Tang, J. ve Rasco, B.A., 2011. Determination of total phenolic content and antioxidant capacity of onion (Allium cepa) and shallot (Allium oschaninii) using infrared spectroscopy, Food Chemistry, 129, 2, 637-644.
• Lyras, L., Cairns, N.J., Jenner, A., Jenner, P. ve Halliwell, B., 1997. An assessment of oxidative damage to proteins, lipids, and DNA in brain from patients with Alzheimer’s disease, Journal of Neurochemical Research, 68, 2061–2069.
• Marnett, L.J., 1999. Lipid peroxidation DNA damage by malondialdehyde, Mutation Research, 424, 83–95.
• Martos, I., Cosentini, M., Ferreres, F. ve Tomas-Barberan, F.A., 1997. Flavonoid Composition of Tunisian Honeys and Propolis. Journal of Agricultural and Food Chemistry, 45, 2824-2829.
• Masuda, T., Yamashita, D., Takeda, Y. ve Yonemori, S., 2005. Screening for tyrosinase inhibitors among extracts for of seashore plants and identification of potent inhibitors from Garcinia subelliptica, Bioscience, Biotechnology, and Biochemistry, 69, 197-201.
• Mohanty, S., Kamolvit, W., Zambrana, S., Sandström, C., Gonzales, E., Ostenson, C.G. ve Braunerf, A., 2017. Extract of Clinopodium bolivianum protects against E. coli invasion of uroepithelial cells, Journal of Ethnopharmacology, 198, 214-220.
• Opalchenova, G. ve Obreshkova, D., 1999. Antibacterial action of extracts of Clinopodium vulgare L. curative plant. Drug Development and Industrial Pharmacy, 25, 323–328.
• Perez, G.R.M., 2013. Antihepatotoxic, nephroprotective, and antioxidant activities of phenolic compounds from Satureja macrostema leaves against carbon tetrachloride-induced hepatic damage in mice, Medicinal Chemistry Research, 22, 1846–1855.
• Sagdıc, O., Kuscu, A., Ozcan, M. ve Ozcelik, S., 2002. Effects of Turkish spice extracts at various concentrations on the growth E. coli O157:H7, Food Microbiology, 19, 473-480.
• Sarikurkcu, C., Ozer, M.S., Tepe, B., Dilek, E. ve Ceylan, O., 2015. Phenolic composition, antioxidant and enzyme inhibitory activities of acetone, methanol and water extracts of Clinopodium vulgare L. subsp. vulgare L. Industrial Crops and Products, 76, 961–966.
• Sayre, L.M., Smith, M.A. ve Perry, G., 2001. Chemistry and biochemistry of oxidative stress in neurodegenerative disease, Current Medicinal Chemistry, 8, 721–738.
• Slinkard, K. ve Singleton, V.L., 1977. Total Phenol Analysis: Automation and Comparison with Manuel Methods. American Journal of Enology and Viticulture, 28, 49-55.
• Soler-Rivas, C., Jolivet, S., Arpin, N., Olivier, J.M. ve Wihers, H.J., 1999. Biochemical and physiological aspects of Brown blotchdisease of Agaricus bisporus. FEMS Microbiology Reviews, 23, 591-614.
• Soumitra, M., Witchuda, K., Silvia, Z., Corine, S., Eduardo, G., Claes-Goran, O. ve Annelie, Br., 2017. Extract of Clinopodium bolivianum protects against E. coli invasion of uroepithelial cells, Journal of Ethnopharmacology, 198,214–220.
• Tepe, B., Sihoglu-Tepe, A., Daferera, D., Polissiou, M. ve Sokmen, A., 2007. Chemical composition and antioxidant activity of the essential oil of Clinopodium vulgare L., Food Chemistry, 103, 766–770.
• Tosun, A., Süntar, İ., Keleş, H., Kiremit, H., Asakawa, Y. ve Akkol, E., 2016. Wound Healing Potential of Selected Liverworts Growing in Turkey, Turkish Journal of Pharmaceutical Sciences, 13, 3, 285-291.
• Turner, B.L., 2008. Taxonomic status of Clinopodium macrostemum (Lamiaceae), Phytologia, 90, 411–413.
• Valko, M., Rhodes, C.J., Moncol, J., Izakovic, M. ve Mazur, M., 2006. Free radicals, metals and antioxidants in oxidative stress-induced cancer, Chemico-Biological Interactions, 160, 1–40.
• Villa-Ruano, N., Pacheco-Hernandez, Y., Zurita-Vasquez, G., Betancourt-Jiménez, M.B., Cruz-Duran, R., Duque-Bautista, H., 2013. Anti-lipase and antioxidant properties of 30 medicinal plants used in Oaxaca México, Biological Research, 46, 153–160.
• Woods, G.L., Brown-Elliott, B.A., Desmond, E.P., Hall, G.S., Heifets, L., Pfyffer, G.E., Ridderhof, J.C., Wallace, R.J., Warren, N.C., Witebsky, F.G. 2003. In Susceptibility Testing of Mycobacteria, Nocardiae, and Other Aerobic Actinomycetes; Approved Standard, NCCLS document M24-A, 23, 18.
• Yarnell, E. ve Abascal, K., 2004. The Leading Publisher in Biotechnology, Alternative Complementary Therapies, 5, 277-284.