ارزیابی تاثیر نوع حلال و شرایط استخراج بر ویژگی‌های کیفی و آنتی‌میکروبی عصاره فاوانیا (Paeonia officinalis L.)
محورهای موضوعی :
بهداشت مواد غذایی
زهرا یعقوبی کوپایی
1
(دانش آموخته کارشناسی ارشد علوم و مهندسی صنایع غذایی، دانشکده کشاورزی و منابع طبیعی، واحد اصفهان (خوراسگان)، دانشگاه آزاد اسلامی، اصفهان، ایران)
هاجر عباسی
2
(دانشیار گروه علوم و مهندسی صنایع غذایی، دانشکده کشاورزی و منابع طبیعی، واحد اصفهان (خوراسگان)، دانشگاه آزاد اسلامی، اصفهان، ایران)
تاریخ دریافت : 1402/01/20
تاریخ پذیرش : 1402/04/27
تاریخ انتشار : 1402/04/01
کلید واژه:
امواج اولتراسونیک,
آنتی‌اکسیدان,
سیال فوق بحرانی,
عودالصلیب,
چکیده مقاله :
گیاه عودالصلیب، با نام علمی Paeonia officinalis L. یکی از گونه‌های متعلق به خانواده Paeoniaceae است. ریشه گیاه عودالصلیب دارای ترکیبات مؤثره متفاوتی از جمله فنل ها ، فلاونوئید ها ، تری‌ترپنوئید ها ،تانن ها، مونوترپن ها و فعالیت های بیولوژیکی متنوعی از جمله اثرات ضد باکتری، ضد اکسیداتیو، آرامش بخش و ضد درد می باشد. در این مطالعه ارزیابی تاثیر سه حلال آب، اتانول/آب، و آب/اتانول/هگزان در استخراج مواد مؤثره ریشه گیاه عودالصلیب در سه روش غوطه وری، غوطه‌وری به‌همراه اعمال امواج اولتراسونیک و استفاده از سیال فوق بحرانی دی‌اکسیدکربن به همراه حلال‌های کمکی مذکور بر راندمان بر ویژگی‌های کیفی (pH، محتوی ترکیبات پلی‌فنولیک و خواص آنتی‌اکسیدانی) و خواص ضدمیکروبی عصاره‌های حاصل بر اشریشیاکلی، استافیلوکوکوس اورئوس، سالمونلا تیفی‌موریوم و کاندیدا آلبیکانس ارزیابی شد. همچنین حداقل غلظت بازدارندگی و کشندگی عصاره ها معین گردید. نتایج نشان داد که نوع حلال مورد استفاده و شرایط استخراج عصاره بر کمیت و کیفیت عصاره‌های تولید شده تاثیر معنی‌داری دارد (05/0p ≤). با استفاده از آب/اتانول و در روش غوطه‌وری با اعمال امواج اولتراسونیک بیش ترین راندمان استخراج عصاره (3/7±0/17) با بالاترین محتوی ترکیبات فنولیک (6/5±0/33) و خاصیت آنتی‌اکسیدانی(4/0±1/76) حاصل شد. حداقل غلظت بازدارندگی و کشندگی این عصاره برای اغلب میکروارگانیسم های مورد مطالعه کمتر از سایر نمونه‌ها بود و قطر هاله عدم رشد اشریشیاکلی، استافیلوکوکوس اورئوس و کاندیدا آلبیکانس در مجاورت این نمونه بیش از سایر عصاره‌ها ارزیابی شد. درحالیکه هاله عدم رشد سالمونلا تیفی‌موریوم در اثر مجاورت با عصاره استخراج شده در آب در روش غوطه‌وری قطر بیشتری داشت. کمیت و کیفیت عصاره مستخرج با سیال فوق بحرانی نسبت به سایر روش‌ها کمتر بود.
چکیده انگلیسی:
Paeonia officinalis L. is one of the species of the Paeoniaceae family. Its root has different effective compounds e.g., polyphenols, flavonoids, triterpenoids, tannins, and monoterpenes. It shows biological activities such as antibacterial, anti-oxidative, relaxing and anti-pain properties. This study evaluated the effect of three solvents, ethanol/water, water, and water/ethanol/hexane on extracting the active ingredients. These solvents were applied by the methods of immersion, immersion with ultrasonic waves, and the use of supercritical fluid carbon dioxide with auxiliary solvents. The efficiency, qualitative characteristics (pH, polyphenolic compounds and antioxidant properties) and antimicrobial properties of the extracts were evaluated by the well-diffusion method against Escherichia coli, Staphylococcus aureus, Salmonella Typhimurium and Candida albicans. Also, the MIC and MBC of the extracts on the activity of these organisms were determined. The results showed that the independent variables significantly affect the quantity and quality of the produced extracts (P ≤ 0.05). Using the water/ethanol, and the immersion method in combination with ultrasonic waves, the highest extract efficiency (17.66± 0.34), of the phenolic compounds (33.5±0.65) and antioxidant property (76±1.44) was obtained. The diameter of the inhibition zone for E. coli, S. aureus and C. albicans in the vicinity of this sample was more than others. Meanwhile, the inhibition zone of S. Typhimurium was larger in the presence of watery extract using the immersion method. The quantity and quality of the extract using supercritical fluid was lower than others.
منابع و مأخذ:
Aliaño-González, M.J., Barea-Sepúlveda, M., Espada-Bellido, E., Ferreiro-González, M., López-Castillo, J.G., Palma, M. et al., (2022). Ultrasound-assisted extraction of total phenolic compounds and antioxidant activity in mushrooms. Agronomy, 12(8): 1812.
Abbas, A., Anwar, F., Ahmad, N., Shahid, M., Al-Mijalli, S. H., Yaseen, M., and Iqbal, M. (2022). Characterization of bioactives and nutra-pharmaceutical potential of supercritical fluid and hydro-distilled extracted coriander leaves essential oil. Dose-Response, 20(4), 15593258221130749.
Ayub, M. A., Choobkar, N., Hanif, M. A., Abbas, M., Ain, Q. U., and Riaz, M. (2022). Chemical composition and biological potential of Pinus roxburghii oleoresin essential oils extracted by steam distillation, superheated steam, and supercritical fluid CO2 extraction. Research Square,
1: 1-18.
Ain Afshar, S. and Sharayi, Parvin. (2020). Investigating the effect of extraction conditions on the amount of bioactive compounds in saffron onion extract. Scientific research of saffron agriculture and technology, 8(1): 89-98 [In Persian]
Ahmad, F. and Tabassum, N. (2013). Preliminary phytochemical, acute oral toxicity and antihepatotoxic study of roots of Paeonia officinalis Linn. Asian Pacific Journal of Tropical Biomedicine, 3(1): 64-68.
Arulmozhi, P., Vijayakumar, S. and Kumar, T. (2018). Phytochemical analysis and antimicrobial activity of some medicinal plants against selected pathogenic microorganisms. Microbial pathogenesis, 123: 219-226.
Delazar, A., Nahar, L., Hamedeyazdan, S. and Sarker, S.D. (2012). Microwave-assisted extraction in natural products isolation. Natural products isolation, 89-115.
Fu, X., Belwal, T., He, Y., Xu, Y., Li, L. and Luo, Z. (2022). UPLC-Triple-TOF/MS characterization of phenolic constituents and the influence of natural deep eutectic solvents on extraction of Carya cathayensis Sarg. Peels: Composition, extraction mechanism and in vitro biological activities. Food Chemistry, 370: 131042.
Gao, L.L., Li, Y.Q., Wang, Z.S., Sun, G.J., Qi, X.M. and Mo, H.Z. (2018). Physicochemical characteristics and functionality of tree peony (Paeonia suffruticosa Andr.) seed protein. Food Chemistry, 240: 980-988.
Abu Najami, M., Ghorbani, M. and Ghorbani Javed, M. (1394). Ultrasonic waves a new method in extracting herbal compounds. Promotional Scientific Journal of Sound and Vibration, 4(8): 119-125. [In Persian]
Hoehn, E., Gasser, F., Guggenbühl, B. and Künsch, U. (2003). Efficacy of instrumental measurements for determination of minimum requirements of firmness, soluble solids, and acidity of several apple varieties in comparison to consumer expectations. Postharvest Biology and Technology, 27(1): 27-37.
Jehanshahi Afshar, A. (2017). Favaniya. Journal of Traditional Medicine of Islam and Iran, 1: 133-138. [In Persian]
Kabiri, S. and Seyed-Alangi, S. Z. (2014). Comparison of antioxidant properties of different extracts of Melissa officinalis leaves obtained by two methods of water extraction and extraction with the help of microwaves and its effect on the oxidation stability of soybean oil. Technology Quarterly New food products, 8: 23-38. [In Persian]
Khaja Nouri, M. and Haghighi, A. (2014). Investigating of natural compounds extraction of plants by microwave and ultrasound waves. Journal of New Food Sciences and Technologies, 1(3): 81-91. [In Persian]
Morelli, L.L.L. and Prado, M.A. (2012). Extraction optimization for antioxidant phenolic compounds in red grape jam using ultrasound with a response surface methodology. Ultrasonics sonochemistry, 19(6): 1144-1149.
Li, P., Shen, J., Wang, Z., Liu, S., Liu, Q., Li, Y. et al., (2021). Genus Paeonia: A comprehensive review on traditional uses, phytochemistry, pharmacological activities, clinical application, and toxicology. Journal of Ethnopharmacology, 269: 113708.
Mokrani, A. and Madani, K. (2016). Effect of solvent, time and temperature on the extraction of phenolic compounds and antioxidant capacity of peach (Prunus persica L.) fruit. Separation and Purification Technology, 162: 68-76.
Milovanovic, I., Zengin, G., Maksimovic, S. and Tadic, V. (2021). Supercritical and ultrasound‐assisted extracts from Pleurotus pulmonarius mushroom: chemical profiles, antioxidative, and enzyme‐inhibitory properties. Journal of the Science of Food and Agriculture, 101(6): 2284-2293.
Mandal, V., Mohan, Y. and Hemalatha, S.J.P.R. (2007). Microwave assisted extraction—an innovative and promising extraction tool for medicinal plant research. Pharmacognosy reviews, 1(1): 7-18.
Moshari-Nasirkandi, A., Alirezalu, A. and Hachesu, M.A. (2020). Effect of lemon verbena bio-extract on phytochemical and antioxidant capacity of strawberry (Fragaria× ananassa Duch. cv. Sabrina) fruit during cold storage. Biocatalysis and Agricultural Biotechnology, 25: 101613.
Nematollahi, A., Mashayekh, M., Sohrabvandi, S., Khosravani, K. and Nejad-Bariki I. (1391). Use of supercritical carbon dioxide in the extraction and purification of vegetable oils. Journal of Nutritional Sciences and Food Industries of Iran, 7(4): 35-44. [In Persian]
Nomanbhay, S., Uskenbayeva, S.A. and Ong, M.Y. (2018). On Increase of The Efficiency of Extracting Phenolic Compounds from Palm Oil Mill Effluent. Journal of Chemical Technology & Metallurgy, 53(1).
Ramak, P., Karimian, V., and Sepahvand, A. (2019). Effects of supercritical fluid and distillation extraction methods on the yield, antioxidant and antifungal activities of Artemisia persica Boiss essential oil. Eco-phytochemical Journal of Medicinal Plants, 7(2), 26-39. [In Persian]
Syukriah, A.N., Liza, M.S., Harisun, Y. and Fadzillah, A.A.M. (2014). Effect of solvent extraction on antioxidant and antibacterial activities from Quercus infectoria (Manjakani). International Food Research Journal, 21(3): 1031.
Saboora, A., Pourbarat, F. and Fallah-Hossini, H. (2014). Comparison of Different Extraction Methods for Optimizing Antioxidant Compounds in Origanum Majorana L. Journal of Shahid Sadoughi University of Medical Sciences, 21: 693-704. [In Persian]
Esmaielzadeh, R., and Raftani Amiri, Z. (2019). The effect of ultrasound assisted and super critical fluid extraction on antioxidant and antibacterial properties of cinnamon extract. Journal of food science and technology (Iran), 16(86), 335-347. [In Persian]
Soria, A.C. and Villamiel, M. (2010). Effect of ultrasound on the technological properties and bioactivity of food: a review. Trends in food science & technology, 21(7): 323-331.
Sharma, S., Barkauskaite, S., Jaiswal, A.K. and Jaiswal, S. (2021). Essential oils as additives in active food packaging. Food Chemistry, 343: 128403.
Shirani Beed Abadi, Kh., Safaiyan, Sh., Mousavi Nadoshan, R. and Rahimi Fard, N. (1400). Investigation of different extraction methods (maceration and ultrasound) on the antioxidant, anti-Alzheimer and antimicrobial properties of Cappadina algae extraction. Journal of Iran Food Sciences and Industries, 112: 69-79. [In Persian]
Soria, A.C. and Villamiel, M. (2010). Effect of ultrasound on the technological properties and bioactivity of food: a review. Trends in food science & technology, 21(7): 323-331.
Sun, X., Guo, X., Ji, M., Wu, J., Zhu, W., Wang, J. et al., (2019). Preservative effects of fish gelatin coating enriched with CUR/βCD emulsion on grass carp (Ctenopharyngodon idellus) fillets during storage at 4 C. Food Chemistry, 272: 643-652.
Tiwari, B.K. (2015). Ultrasound: A clean, green extraction technology. TrAC Trends in Analytical Chemistry, 71: 100-109.
Vardanega, R., Santos, D.T. and Meireles, M.A.A. (2014). Intensification of bioactive compounds extraction from medicinal plants using ultrasonic irradiation. Pharmacognosy reviews, 8(16): 88.
Vega-Arroy, D.J., Ruiz-Espinosa, H., Luna-Guevara, J.J., Luna-Guevara, M.L., Hernandez-Carranza, P., Avila-Sosa, R. et al., (2017). Effect of solvents and extraction methods on total anthocyanins, phenolic compounds and antioxidant capacity of Renealmia alpinia (Rottb.) Maas peel. Czech Journal of Food Sciences, 35(5): 456-465.
Vagi, E., Simándi, B., Daood, H.G., Deak, A. and Sawinsky, J. (2002). Recovery of pigments from Origanum majorana L. by extraction with supercritical carbon dioxide. Journal of agricultural and food chemistry, 50(8): 2297-2301.
Vidović, S., Vasić, A., Vladić, J., Jokić, S., Aladić, K., Gavarić, A. et al., (2021). Carbon dioxide supercritical fluid extracts from yarrow and rose hip herbal dust as valuable source of aromatic and lipophilic compounds. Sustainable Chemistry and Pharmacy, 22: 100494.
Vinatoru, M. (2001). An overview of the ultrasonically assisted extraction of bioactive principles from herbs. Ultrasonics sonochemistry, 8(3): 303-313.
Yan, Z., Xie, L., Li, M., Yuan, M., Tian, Y., Sun, D. et al., (2021). Phytochemical components and bioactivities of novel medicinal food–peony roots. Food Research International, 140: 109902.
Yeasmin, D., Swarna, R.J., Nasrin, M.S., Parvez, S. and Alam, M.F. (2016). Evaluation of antibacterial activity of three flower colours Chrysanthemum morifolium Ramat. Against multi-drug resistant human pathogenic bacteria. International Journal of Biosciences, 9: 78-87.
_||_
Aliaño-González, M.J., Barea-Sepúlveda, M., Espada-Bellido, E., Ferreiro-González, M., López-Castillo, J.G., Palma, M. et al., (2022). Ultrasound-assisted extraction of total phenolic compounds and antioxidant activity in mushrooms. Agronomy, 12(8): 1812.
Abbas, A., Anwar, F., Ahmad, N., Shahid, M., Al-Mijalli, S. H., Yaseen, M., and Iqbal, M. (2022). Characterization of bioactives and nutra-pharmaceutical potential of supercritical fluid and hydro-distilled extracted coriander leaves essential oil. Dose-Response, 20(4), 15593258221130749.
Ayub, M. A., Choobkar, N., Hanif, M. A., Abbas, M., Ain, Q. U., and Riaz, M. (2022). Chemical composition and biological potential of Pinus roxburghii oleoresin essential oils extracted by steam distillation, superheated steam, and supercritical fluid CO2 extraction. Research Square,
1: 1-18.
Ain Afshar, S. and Sharayi, Parvin. (2020). Investigating the effect of extraction conditions on the amount of bioactive compounds in saffron onion extract. Scientific research of saffron agriculture and technology, 8(1): 89-98 [In Persian]
Ahmad, F. and Tabassum, N. (2013). Preliminary phytochemical, acute oral toxicity and antihepatotoxic study of roots of Paeonia officinalis Linn. Asian Pacific Journal of Tropical Biomedicine, 3(1): 64-68.
Arulmozhi, P., Vijayakumar, S. and Kumar, T. (2018). Phytochemical analysis and antimicrobial activity of some medicinal plants against selected pathogenic microorganisms. Microbial pathogenesis, 123: 219-226.
Delazar, A., Nahar, L., Hamedeyazdan, S. and Sarker, S.D. (2012). Microwave-assisted extraction in natural products isolation. Natural products isolation, 89-115.
Fu, X., Belwal, T., He, Y., Xu, Y., Li, L. and Luo, Z. (2022). UPLC-Triple-TOF/MS characterization of phenolic constituents and the influence of natural deep eutectic solvents on extraction of Carya cathayensis Sarg. Peels: Composition, extraction mechanism and in vitro biological activities. Food Chemistry, 370: 131042.
Gao, L.L., Li, Y.Q., Wang, Z.S., Sun, G.J., Qi, X.M. and Mo, H.Z. (2018). Physicochemical characteristics and functionality of tree peony (Paeonia suffruticosa Andr.) seed protein. Food Chemistry, 240: 980-988.
Abu Najami, M., Ghorbani, M. and Ghorbani Javed, M. (1394). Ultrasonic waves a new method in extracting herbal compounds. Promotional Scientific Journal of Sound and Vibration, 4(8): 119-125. [In Persian]
Hoehn, E., Gasser, F., Guggenbühl, B. and Künsch, U. (2003). Efficacy of instrumental measurements for determination of minimum requirements of firmness, soluble solids, and acidity of several apple varieties in comparison to consumer expectations. Postharvest Biology and Technology, 27(1): 27-37.
Jehanshahi Afshar, A. (2017). Favaniya. Journal of Traditional Medicine of Islam and Iran, 1: 133-138. [In Persian]
Kabiri, S. and Seyed-Alangi, S. Z. (2014). Comparison of antioxidant properties of different extracts of Melissa officinalis leaves obtained by two methods of water extraction and extraction with the help of microwaves and its effect on the oxidation stability of soybean oil. Technology Quarterly New food products, 8: 23-38. [In Persian]
Khaja Nouri, M. and Haghighi, A. (2014). Investigating of natural compounds extraction of plants by microwave and ultrasound waves. Journal of New Food Sciences and Technologies, 1(3): 81-91. [In Persian]
Morelli, L.L.L. and Prado, M.A. (2012). Extraction optimization for antioxidant phenolic compounds in red grape jam using ultrasound with a response surface methodology. Ultrasonics sonochemistry, 19(6): 1144-1149.
Li, P., Shen, J., Wang, Z., Liu, S., Liu, Q., Li, Y. et al., (2021). Genus Paeonia: A comprehensive review on traditional uses, phytochemistry, pharmacological activities, clinical application, and toxicology. Journal of Ethnopharmacology, 269: 113708.
Mokrani, A. and Madani, K. (2016). Effect of solvent, time and temperature on the extraction of phenolic compounds and antioxidant capacity of peach (Prunus persica L.) fruit. Separation and Purification Technology, 162: 68-76.
Milovanovic, I., Zengin, G., Maksimovic, S. and Tadic, V. (2021). Supercritical and ultrasound‐assisted extracts from Pleurotus pulmonarius mushroom: chemical profiles, antioxidative, and enzyme‐inhibitory properties. Journal of the Science of Food and Agriculture, 101(6): 2284-2293.
Mandal, V., Mohan, Y. and Hemalatha, S.J.P.R. (2007). Microwave assisted extraction—an innovative and promising extraction tool for medicinal plant research. Pharmacognosy reviews, 1(1): 7-18.
Moshari-Nasirkandi, A., Alirezalu, A. and Hachesu, M.A. (2020). Effect of lemon verbena bio-extract on phytochemical and antioxidant capacity of strawberry (Fragaria× ananassa Duch. cv. Sabrina) fruit during cold storage. Biocatalysis and Agricultural Biotechnology, 25: 101613.
Nematollahi, A., Mashayekh, M., Sohrabvandi, S., Khosravani, K. and Nejad-Bariki I. (1391). Use of supercritical carbon dioxide in the extraction and purification of vegetable oils. Journal of Nutritional Sciences and Food Industries of Iran, 7(4): 35-44. [In Persian]
Nomanbhay, S., Uskenbayeva, S.A. and Ong, M.Y. (2018). On Increase of The Efficiency of Extracting Phenolic Compounds from Palm Oil Mill Effluent. Journal of Chemical Technology & Metallurgy, 53(1).
Ramak, P., Karimian, V., and Sepahvand, A. (2019). Effects of supercritical fluid and distillation extraction methods on the yield, antioxidant and antifungal activities of Artemisia persica Boiss essential oil. Eco-phytochemical Journal of Medicinal Plants, 7(2), 26-39. [In Persian]
Syukriah, A.N., Liza, M.S., Harisun, Y. and Fadzillah, A.A.M. (2014). Effect of solvent extraction on antioxidant and antibacterial activities from Quercus infectoria (Manjakani). International Food Research Journal, 21(3): 1031.
Saboora, A., Pourbarat, F. and Fallah-Hossini, H. (2014). Comparison of Different Extraction Methods for Optimizing Antioxidant Compounds in Origanum Majorana L. Journal of Shahid Sadoughi University of Medical Sciences, 21: 693-704. [In Persian]
Esmaielzadeh, R., and Raftani Amiri, Z. (2019). The effect of ultrasound assisted and super critical fluid extraction on antioxidant and antibacterial properties of cinnamon extract. Journal of food science and technology (Iran), 16(86), 335-347. [In Persian]
Soria, A.C. and Villamiel, M. (2010). Effect of ultrasound on the technological properties and bioactivity of food: a review. Trends in food science & technology, 21(7): 323-331.
Sharma, S., Barkauskaite, S., Jaiswal, A.K. and Jaiswal, S. (2021). Essential oils as additives in active food packaging. Food Chemistry, 343: 128403.
Shirani Beed Abadi, Kh., Safaiyan, Sh., Mousavi Nadoshan, R. and Rahimi Fard, N. (1400). Investigation of different extraction methods (maceration and ultrasound) on the antioxidant, anti-Alzheimer and antimicrobial properties of Cappadina algae extraction. Journal of Iran Food Sciences and Industries, 112: 69-79. [In Persian]
Soria, A.C. and Villamiel, M. (2010). Effect of ultrasound on the technological properties and bioactivity of food: a review. Trends in food science & technology, 21(7): 323-331.
Sun, X., Guo, X., Ji, M., Wu, J., Zhu, W., Wang, J. et al., (2019). Preservative effects of fish gelatin coating enriched with CUR/βCD emulsion on grass carp (Ctenopharyngodon idellus) fillets during storage at 4 C. Food Chemistry, 272: 643-652.
Tiwari, B.K. (2015). Ultrasound: A clean, green extraction technology. TrAC Trends in Analytical Chemistry, 71: 100-109.
Vardanega, R., Santos, D.T. and Meireles, M.A.A. (2014). Intensification of bioactive compounds extraction from medicinal plants using ultrasonic irradiation. Pharmacognosy reviews, 8(16): 88.
Vega-Arroy, D.J., Ruiz-Espinosa, H., Luna-Guevara, J.J., Luna-Guevara, M.L., Hernandez-Carranza, P., Avila-Sosa, R. et al., (2017). Effect of solvents and extraction methods on total anthocyanins, phenolic compounds and antioxidant capacity of Renealmia alpinia (Rottb.) Maas peel. Czech Journal of Food Sciences, 35(5): 456-465.
Vagi, E., Simándi, B., Daood, H.G., Deak, A. and Sawinsky, J. (2002). Recovery of pigments from Origanum majorana L. by extraction with supercritical carbon dioxide. Journal of agricultural and food chemistry, 50(8): 2297-2301.
Vidović, S., Vasić, A., Vladić, J., Jokić, S., Aladić, K., Gavarić, A. et al., (2021). Carbon dioxide supercritical fluid extracts from yarrow and rose hip herbal dust as valuable source of aromatic and lipophilic compounds. Sustainable Chemistry and Pharmacy, 22: 100494.
Vinatoru, M. (2001). An overview of the ultrasonically assisted extraction of bioactive principles from herbs. Ultrasonics sonochemistry, 8(3): 303-313.
Yan, Z., Xie, L., Li, M., Yuan, M., Tian, Y., Sun, D. et al., (2021). Phytochemical components and bioactivities of novel medicinal food–peony roots. Food Research International, 140: 109902.
Yeasmin, D., Swarna, R.J., Nasrin, M.S., Parvez, S. and Alam, M.F. (2016). Evaluation of antibacterial activity of three flower colours Chrysanthemum morifolium Ramat. Against multi-drug resistant human pathogenic bacteria. International Journal of Biosciences, 9: 78-87.
