بررسی ترکیبات شیمیایی، گروه‏های عاملی زیست‏فعال، قدرت آنتی‌اکسیدانی و سمیت سلولی اسانس آویشن شیرازی بر رده سلولیHT29: مطالعه در شرایط آزمایشگاهی

نوع مقاله : مقاله پژوهشی

نویسندگان

• الناز صفاری سامانی ¹
• حسین جوینده ²
• بهروز علیزاده بهبهانی ³

¹ دانشجوی کارشناسی ارشد، گروه علوم و مهندسی صنایع غذایی، دانشکده علوم دامی و صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی خوزستان، ملاثانی، ایران.

² استاد، گروه علوم و مهندسی صنایع غذایی، دانشکده علوم دامی و صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی خوزستان، ملاثانی، ایران

³ گروه علوم و مهندسی صنایع غذایی، دانشکده علوم دامی و صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی خوزستان، ملاثانی، ایران

https://doi.org/10.22067/ifstrj.2020.40161.0

چکیده

هدف از این پژوهش شناسایی ترکیبات شیمیایی، گروه‏های عاملی زیست‏فعال، فعالیت آنتی‌اکسیدانی، فنل کل و فلاونوئید کل اسانس آویشن شیرازی و همچنین بررسی اثر سمیت سلولی آن بر رده سلولی سرطان روده بزرگ (HT29) بود. ترکیبات شیمیایی اسانس آویشن شیرازی با استفاده از دستگاه کروماتوگرافی گازی متصل به طیف‌سنج جرمی شناسایی شد. گروه‏های عاملی زیست‏فعال اسانس آویشن شیرازی با استفاده از دستگاه FTIR در محدوده عدد موجی
cm^-1 500-4000 اندازه‌گیری شد. به‌منظور اندازه‌گیری میزان فنل کل و فلاونوئید کل اسانس آویشن شیرازی به‌ترتیب از روش‌های فولین سیوکالتو و رنگ‌سنجی آلومینیوم کلراید استفاده گردید. از روش MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyltetrazolium bromide) جهت اندازه‌گیری سمیت سلولی اسانس آویشن شیرازی در برابر رده سلولی سرطان روده بزرگ (HT29) استفاده شد. قدرت آنتی‌کسیدانی اسانس آویشن شیرازی با استفاده از مهار رادیکال آزاد DPPH و ABTS بررسی گردید. ترکیب کارواکرول و تیمول به‌ترتیب با 3/39 و 30 درصد بیشترین ترکیب تشکیل‌دهنده اسانس آویشن شیرازی بودند. وجود پیک در عدد موجی cm ^-11000-320 نشان‌دهنده پیوندهای C-O است که می‌تواند مربوط به ترکیبات آلی همچون الکل‌ها، اسیدهای کربوکسیلیک، استرها و اتر باشد. میزان فنل کل و فلاونوئید اسانس آویشن شیرازی به‌ترتیب 05/64 میلی‌گرم گالیک اسید بر گرم و 68/11 میلی‌گرم کوئرستین بر گرم بود. میزان درصد مهارکنندگی رادیکال‌های آزاد با استفاده از DPPH و ABTS در غلظت ppm 1000 به‌ترتیب 69/63 و 33/64 بود. با افزایش غلظت اسانس آویشن شیرازی، تأثیر بر رده سلولی HT29 افزایش پیدا کرد و درصد زنده‌مانی آن کاهش یافت. با توجه به نتایج قدرت آنتی‌اکسیدانی، فنل و فلاونوئید کل اسانس آویشن شیرازی به نظر می‌رسد بتوان از گیاه آویشن شیرازی به‌عنوان نگهدارنده طبیعی در صنایع غذایی بهره برد.

کلیدواژه‌ها

• اسانس آویشن شیرازی
• سمیت سلولی
• فعالیت آنتی‌اکسیدانی
• زنده‌مانی سلول‌های HT29

عنوان مقاله [English]

Investigation on the chemical composition, bioactive functional groups, antioxidant potential and cell toxicity (HT29) of Shirazi thyme essential oil: A study in laboratory scale

نویسندگان [English]

• Elnaz Saffari Samani ¹
• Hossein Jooyandeh ²
• Behrooz Alizadeh Behbahani ³

¹ MSc. student, Department of Food Science and Technology, Faculty of Animal Science and Food Technology, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Iran.

² Professor, Department of Food Science and Technology, Faculty of Animal Science and Food Technology, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Iran

³ Department of Food Science and Technology, Faculty of Animal Science and Food Technology, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Iran

چکیده [English]

Introduction: Shirazi thyme is a perennial herbaceous plant related to Lamiaceae family growing in central and southern regions of Iran. This study aimed to assess chemical composition, bioactive functional groups, antioxidant potential, total phenol, and total flavonoids of Shirazi thyme essential oil (STEO) and also to evaluate its cell toxicity effect against the colorectal cancer cell line (HT29).
 
Materials and Methods: The chemical composition of STEO was identified by gas chromatography-mass spectrometry. The bioactive functional groups of STEO were measured by Fourier transform infrared at spectra range of 500- 4000 cm^-1. The total phenol and total flavonoids of STEO were determined by folin- ciocalteu and aluminum chloride colorimetric method, respectively. MMT method was performed to measure the cell toxicity of STEO against the colorectal cancer cell line (HT29). The cells were cultured on DMEM high glucose medium supplemented with 10% fetal bovine serum and penicillin/streptomycin and incubated at 37°C with 95% relative humidity and 5% CO₂ concentration. The antioxidant activity of STEO was evaluated by DPPH and ABTS free radicals scavenging assays. Duncan test at 5% probability and SPSS software (version 18) was performed to compare the means of obtained results.
 
Results and Discussion: Based on GC/MS spectrometry, a total of 29 constituents were identified and quantified in the STEO representing more than 99% of total constituents. Thymol with 39.3% and Carvacrol with 30% were the most constituents of STEO. The other major compound of STEO were Benzene (8.52%), γ-Terpinene (5.27%), and Caryophyllene (2.97%). The presence of peak at a wavelength of 320- 1000 cm^-1 demonstrates O-C bound that could be related to organic compounds such as alcohols, carboxylic acids, esters, and ethers. The peaks occurred at spectral range between 2800- 3000 cm^-1 (particularly at 2869.6 up to 2960.8 cm^-1) are related to the stretching mode of C-H bounds and are mostly associated with alcoholic compounds in the essence. The amount of total phenol and flavonoids of STEO were 64.05 mg gallic acid equivalent/g and 11.68 mg quercetin equivalent/g, respectively. Results obtained from antioxidant activity of STEO with DPPH and ABTS free radicals scavenging showed that by increasing of STEO, the inhibition level of free radicals was enhanced. The inhibition percent of free radicals by using DPPH and ABTS at 1000 ppm concentration were 63.69% and 64.33%, respectively. The cell toxicity results of different STEO concentrations against the cell line HT29 revealed that survival of HT29 cells at 1, 3.125, 6.25, 12.5, 25, 50, 100 and 200 ppm of STEO were 100, 70.15, 61.19, 59.33, 38.83, 25.68, 20.65 and 12.2%. According to the results of this study, as the concentration of STEO increased, its effect on the cell line HT29 enhanced and the cell percentage viability decreased. Some antioxidant activity of STEO could be interrelated to its phenolic compounds. Based on the results of antioxidant activity, the amount of total phenol and flavonoids and the effect of STEO on cell toxicity against the cell line HT29, it seems that the use of STEO is applicable in the pharmaceutical and food industries

کلیدواژه‌ها [English]

• Shirazi thyme essential oil
• Cell toxicity
• Antioxidant activity
• HT29 cell viability

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