ارزﯾﺎﺑﯽ ﺧﻮاص آﻧﺘﯽ اﮐﺴﯿﺪاﻧﯽ و ﺿﺪ ﺑﺎﮐﺘﺮﯾﺎﯾﯽ ﮐﻠﺮوﻓﯿﻞ اﺳﺘﺨﺮاج ﺷﺪه از ﮔﯿﺎه ﺷﺒﺪر اﯾﺮاﻧﯽ (.Trifolium resupinatum L)

Evaluation of Antioxidant and Antibacterial Properties of Chlorophyll Extracted from Persian Clover (Trifolium resupinatum L.)

ﺑﻪ ﮐﺎرﮔﯿﺮي رﻧﮓﻫﺎي ﻃﺒﯿﻌﯽ در ﻣﻮاد ﻏﺬاﯾﯽ ﻣﻮﺟﺐ اﻓﺰاﯾﺶ ﮐﯿﻔﯿﺖ و وﯾﮋﮔﯽﻫﺎي ﺣﺴﯽ ﻣﺤﺼﻮل ﺷﺪه و ﺑﺎ اﺛﺮات ﺿﺪاﻟﺘﻬﺎﺑﯽ و ﺿﺪ ﺳﺮﻃﺎﻧﯽ ﻧﯿﺰ ﻫﻤﺮاه ﻣﯽﺑﺎﺷﻨﺪ و ﺑﻨﺎﺑﺮاﯾﻦ، ﻧﻘﺶ ﻣﻬﻤﻰ در ﺳﻼﻣﺖ اﻧﺴﺎن ﺑﺮ ﻋﻬﺪه دارﻧﺪ. ﮐﻠﺮوﻓﯿﻞ ﻧﯿﺰ ﺑﻪ ﻋﻨﻮان ﯾﮑﯽ از رﻧﮓ داﻧﻪ ﻫﺎي ﻃﺒﯿﻌﯽ، در اﻧﻮاع ﻣﻮاد ﻏﺬاﯾﯽ ﻣﻮرد اﺳﺘﻔﺎده ﻗﺮار ﻣﯽﮔﯿﺮد. ﻫﺪف از اﯾﻦ ﺗﺤﻘﯿﻖ اﺳﺘﻔﺎده از روش آﻧﺰﯾﻤﯽ ﺑﻪ ﻣﻨﻈﻮر اﺳﺘﺨﺮاج ﮐﻠﺮوﻓﯿﻞ از ﮔﯿﺎه ﺷﺒﺪر اﯾﺮاﻧﯽ و ارزﯾﺎﺑﯽ ﺧﻮاص آﻧﺘﯽ اﮐﺴﯿﺪاﻧﯽ و وﯾﮋﮔﯽﻫﺎي ﺿﺪ ﺑﺎﮐﺘﺮﯾﺎﯾﯽ آن ﺑﻮده اﺳﺖ. ﻓﻌﺎﻟﯿﺖ آﻧﺘﯽاﮐﺴﯿﺪاﻧﯽ ﮐﻠﺮوﻓﯿﻞ اﺳﺘﺨﺮاج ﺷﺪه از ﮔﯿﺎه ﺷﺒﺪر اﯾﺮاﻧﯽ ﺑﺎ ﺳﻪ روش ﻣﻬﺎر رادﯾﮑﺎلﻫﺎي آزاد DPPH و ABTS و ﻗﺪرت اﺣﯿﺎﮐﻨﻨﺪﮔﯽ آﻫﻦ )FRAP( و اﺛﺮات ﺿﺪ ﺑﺎﮐﺘﺮﯾﺎﯾﯽ ﻋﻠﯿﻪ ﻣﯿﮑﺮوارﮔﺎﻧﯿﺴﻢﻫﺎي ﻣﺨﺘﻠﻒ، ﺑﺎ دو روش اﻧﺘﺸﺎر دﯾﺴﮏ در ﻣﺤﯿﻂ آﮔﺎردار و رﻗﯿﻖ ﺳﺎزي )MIC و MBC( اﻧﺠﺎم ﮔﺮﻓﺖ. ﻧﺘﺎﯾﺞ ﻧﺸﺎن داد ﮐﻪ ﻏﻠﻈﺖ ﮐﻠﺮوﻓﯿﻞ a در ﺷﺒﺪر اﯾﺮاﻧﯽ ﺗﻘﺮﯾﺒﺎً دو ﺑﺮاﺑﺮ ﻏﻠﻈﺖ ﮐﻠﺮوﻓﯿﻞ b ﺑﻮده و ﻏﻠﻈﺖﻫﺎي ﺑﺎﻻﺗﺮ ﮐﻠﺮوﻓﯿﻞ ﻓﻌﺎﻟﯿﺖ آﻧﺘﯽاﮐﺴﯿﺪاﻧﯽ ﺑﯿﺸﺘﺮي در ﻣﻬﺎر رادﯾﮑﺎلﻫﺎي آزاد DPPH و ABTS و ﻧﯿﺰ ﻗﺪرت اﺣﯿﺎﮐﻨﻨﺪﮔﯽ آﻫﻦ ﻧﺸﺎن دادﻧﺪ )0/05 < p(. اﺛﺮات ﺿﺪ ﺑﺎﮐﺘﺮﯾﺎﯾﯽ ﮐﻠﺮوﻓﯿﻞ در ﻏﻠﻈﺖﻫﺎي 20، 40، 60 و 100 ﻣﯿﮑﺮوﻣﻮﻻر ﻣﺘﻔﺎوت ﺑﻮده ﺑﻪ ﻃﻮري ﮐﻪ ﻣﻘﺎوﻣﺖ ﺑﺎﮐﺘﺮيﻫﺎ در روش اﻧﺘﺸﺎر ﺑﻪ ﺗﺮﺗﯿﺐ ﺳﻮدوﻣﻮﻧﺎس< اﺷﺮﺷﯿﺎ<ﺳﺎﻟﻤﻮﻧﻼ< اﺳﺘﺎﻓﯿﻠﻮﮐﻮﮐﻮس< ﻟﯿﺴﺘﺮﯾﺎ )0/05 < p( و در روش رﻗﯿﻖ ﺳﺎزي ﺑﻪ ﺗﺮﺗﯿﺐ ﺳﻮدوﻣﻮﻧﺎس< ﺳﺎﻟﻤﻮﻧﻼ< اﺷﺮﺷﯿﺎ< اﺳﺘﺎﻓﯿﻠﻮﮐﻮﮐﻮس= ﻟﯿﺴﺘﺮﯾﺎ ﻣﺸﺎﻫﺪه اﺳﺖ. ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﻧﺘﺎﯾﺞ ﺑﻪ دﺳﺖ آﻣﺪه و ﺗﺄﺛﯿﺮات آﻧﺘﯽاﮐﺴﯿﺪاﻧﯽ و ﺿﺪ ﺑﺎﮐﺘﺮﯾﺎﯾﯽ ﮐﻠﺮوﻓﯿﻞ در ﻏﻠﻈﺖﻫﺎي ﺑﺎﻻﺗﺮ، ﻣﯽﺗﻮان از آن ﺑﻪ ﻋﻨﻮان رﻧﮓ داﻧﻪ ﺷﺎﺧﺺ در ﻣﻮاد ﻏﺬاﯾﯽ و داروﯾﯽ اﺳﺘﻔﺎده ﻧﻤﻮد. وﻟﯽ ﺑﺎ اﯾﻦ وﺟﻮد، اﻧﺠﺎم آزﻣﺎﯾﺶ ﻫﺎي ﺗﮑﻤﯿﻠﯽ ﺑﻪ ﻣﻨﻈﻮر دﺳﺘﯿﺎﺑﯽ ﺑﻪ ﻧﺘﺎﯾﺞ ﺑﯿﺸﺘﺮ ﺿﺮوري ﺑﻪ ﻧﻈﺮ ﻣﯽرﺳﺪ.

The use of natural dyes in foods enhances the quality, efficiency and sensory properties of the product and also, they are associated with anti-inflammatory and anti-cancer effects and therefore they play an important role in human health. Chlorophyll is used as a natural pigment in a variety of foods. The aim of this study was extraction of chlorophyll from Persian clover (Trifolium resupinatum L.) using enzyme method and evaluation its antioxidant and antibacterial properties. Antioxidant activity of chlorophyll extracted from Persian clover was performed by three methods including DPPH and ABTS free radicals scavenging and ferric reducing antioxidant power (FRAP) and antibacterial effects against different microorganisms was also carried out using agar disk diffusion and microdilution (MIC and MBC) methods. The results showed that the concentration of chlorophyll a was almost twice of chlorophyll b and higher concentrations of chlorophyll had higher antioxidant activity in DPPH and ABTS free radicals scavenging and FRAP (p>0.05). Antibacterial effects of chlorophyll at concentrations of 20, 40, 60 and 100 μM were different, with bacterial resistance in Pseudomonas aeruginosa > Salmonella typhi > Escherichia coli > Staphylococcus aureus > Listeria monocytogenes (P<0.05) in agar disk diffusion and Pseudomonas aeruginosa > Salmonella typhi > Escherichia coli > Staphylococcus aureus = Listeria monocytogenes in microdilution respectively. Based on the results and the antioxidant and antibacterial effects of chlorophyll at higher concentrations, it can be used as a biocolorant in food and pharmaceuticals. However, further tests are needed to obtain further results.