2010,32 (1) :24 28 http :/ / xuebao. jlau. edu. cn Journal of Jilin Agricultural University E2mail : jlndxb @vip. sina. com Ξ,,,, ΞΞ, 200062 : : 320 mg/ L,; ph 4 9, ; 80, 100,121 : : ; ; : TS20213 : A : 100025684 (2010) 0120024205 Antimicrobial Ability of Limonene, a Natural and Active Monoterpene WANG Xue2mei, CHEN Hui, LI Xue2jiao, SUN Yong2hong, GAO Hong2liang School of Life Science, East China Normal University, Shanghai 200062, China Abstract : The antimicrobial activity of limonene was determined by minimum inhibitory concentration (MIC) and inhibitory circle. The results showed that MIC to and sp. was 320 mg/ L. However, it didn t show obvious antimicrobial effect on Staphylococcus aureus. Its antimicrobial ability was stable in ph value between 4 and 9. It was also stable when heated at 80. However, the antimicrobial ability to sp. and decreased when heated at 100. And it decreased when heated at 121. The results of inhibitory circle showed that limonene had better antimicrobial ability to, sp. and Aspergillus niger than potassium sorbate and sodium benzoate. Key words : limonene ; antimicrobial ability ; food preservative,12242(12 ),C 10 H 16,,, [1 ], GRAS ( ), FDA, FAO WHO ADI [2 ],,,, [3 ], [4 ], Ξ ΞΞ : (30600079) :,,,: : 2008212214 : 2009203213
: 25, 1 111, ( ) ( Staphylococcus aureus) ( Bacillus subtilis) ( Bacillus ) ( Aspergillus niger) ( Penicillum sp1) ( ) ( sp1) 112 752 SLS - 60-70 ( ) BME100L ( ) 113 [5 ], [6 ] 114 100 g 20 g, 1 000 ml 80 ;,5 000 r/ min,10 min ; SLS - 60-70 5,20 MPa 115 11511 (10 ml),,40,80, 160, 320, 640, 1 000, 2 000, 3 000, 4 000, 5 000 mg/ L 100 L () 10 7 cfu/ ml 28 72 h 28 48 h37 24 h, (MIC) [728 ] 11512 MIC(MIC) 1 ml,11511, (MBC) [728 ] 11513 ph 0110 mol/ L 0110 mol/ L NaOH phph 4,5,6,7,8,9 6, MIC [9 ] (11511 ) 11514 80 100 121 15 min, MIC [9 ] (11511 ) 11515 1 ml, 0110 ml (10 6 cfu), 1, 3, 6, 12 h, := ( - ) / 100 % 11516 20 ml,5 ml 50 012 ml () 10 7 cfu/ ml, 6 mm,10 g/ L, [10 ] 11511 2 2. 1 1, MIC, MBC MIC 3 000,2 000,1 000, 320,320,4 000 mg/ L, MBC 2 000,640, 640,4 000 mg/ L,,J. S. Dambolena [4 ],, Journal of Jilin Agricultural University
2 6 2010 2 1 Table 1. MIC and MBC of limonene on different microorganisms ck () / (mg L - 1 ) 40 80 160 320 640 1 000 2 000 3 000 4 000 5 000 MIC/ (mg L - 1 ) MBC/ (mg L - 1 ) Staphylococcus aureus Bacillus sp1 Penicillum sp1 Aspergillus niger +++ +++ +++ +++ +++ +++ +++ + 3 000 > 5 000 +++ +++ +++ +++ +++ +++ + 2 000 > 5 000 +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ > 5 000 > 5 000 +++ +++ +++ +++ ++ + 1 000 2 000 +++ +++ + + 320 640 +++ +++ ++ + 320 640 +++ +++ +++ +++ +++ +++ +++ +++ ++ ++ ++ > 5 000 > 5 000 +++ +++ +++ +++ +++ +++ +++ +++ ++ 4 000 4 000 Note : Absent growth ; + Mild growth ; ++ Strong growth ; +++ Intense growth,the same below 2. 2 ph 2, ph 4 9,, ph,, [11212 ] 2 ph Table 2. Effect of ph on antimicrobial effect of limonene Bacillus sp1 Aspergillus niger ph 4 5 6 7 8 9 C ck C ck C ck C ck C ck C ck + ++ +++ +++ +++ +++ + +++ +++ +++ +++ +++ +++ +++ +++ : C MIC Note : C contains the same concentration of limonene with MIC 2. 3 3, 80 15 min, 100 15 min, 121 15 min [1 ], Journal of Jilin Agricultural University 2010, February
: 27 3 Table 3. Antimicrobial thermal stability of limonene ck 80 100 121 +++ + + +++ Bacillus +++ + +++ + sp. +++ + ++ Aspergillus niger +++ + 2. 4,,1 2,,,1 1 Fig. 1. Relationship between antimicrobial rate and treating time of limonene to Bacillus 1 2,,,,, 2 Fig. 2. Relationship between antimicrobial rate and treating time of limonene to Saccharomyce s 2. 5,,44, 4 ( ) Table 4. Comparison of antimicrobial effect between limonene and chemical pre servative ( Inhibitory circle diameter) Agent Bacillus Saccharomyc2 es sp1 ck 0 0 0 0 0 0 Limonene Potassium sorbate Sodium benzoate Aspergillus niger 9162 10151 13151 15151 14155 9133 9163 9154 13122 10184 10152 8151 9159 9154 13129 9129 9118 8132 mm Journal of Jilin Agricultural University
2 8 2010 2 3 1),,; 2) ph, 3) 80, 100, 121, 4) 5), : [1 ],. [J ]., 2007 (16) : 64. [2 ]. [J ]., 2005 (1) : 33237. [3 ]. [J ]., 2006, 27 (3) : 1622165. [4 ] Dambolena J S, L pez A G, Cgnepa M C, et al. Inhibitory effect of cyclic terpenes (limonene, menthol, menthone and thymol) on Fusarium verticillioides MRC 826 growth and fumonisin B1 biosyn2 thesis[j ]. Toxicon, 2008, 51 : 37244. [5 ],,. [M]. :, 1999. [6 ],,,. [J ]., 2008, 37 (5) : 31235. [7 ] Rasooli I, Rezaei M B, Allameh A. Ultrastructural studies on an2 timicrobial efficacy of thyme essential oils on Listeria monocytogenes [J ]. International Journal of Infectious Diseases, 2006, 10 : 2362 241. [8 ] Rasooli I, Owlia P. Chemoprevention by thyme oils of Aspergillus parasiticus growth and aflatoxin production [ J ]. Phytochemistry, 2005, 66 : 285122856. [9 ],,. [J ]., 2005, 25 (1) : 56259. [10 ],,. 8 [J ]., 2008,13 (1) :9210. [11 ],,. [J ]., 2008, 29 (3) : 1572161. [12 ],,, 1 [J ]1,2008,30 (4) :5962604. (16 ) 4 411 4,60 kg/ hm 2,16 18 / hm 2 150 kg/ hm 2,30 t/ hm 2 412 4,60 65 cm, 10 cm,18 20 / hm 2,, 413 24 : [1 ],,,. 12 [J ].,2003,25 (1) :123. [2 ],,,. 14 [J ].,2004,26 (2) :1192121. [3 ],,,. 16 [J ].,2006,28 (1) :10212. [4 ],,,. 19 [J ].,2007,29 (3) :2472249. [5 ],,,. 13 [J ].,2004,26 (1) :13215. [6 ],,,. 13 [J ].,2002,24 (6) :123. [7 ],,, 1 23 [J ].,2008,30 (6) :7792781,788. Journal of Jilin Agricultural University 2010, February