Abstract: Objective To examine and identify fungi with important significance in food safety by matrix2assisted laser

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1 (11, ; 21, ) :(MALDI2TOF2MS) MALDI2TOF2MS,, MALDI2TOF2MS, MBT; IAA ;Π1 1 ; MALDI2TOF2MS :,, ; ;, Detection and Identification of Fungi by Matrix2Assisted Laser DesorptionΠIonization Time of Flight Mass Spectrometry LI Feng2qin, WU Duo2jia, WU Shu2zhen, LI Yu2wei (National Institute for Nutrition and Food Safety, Chinese CDC, Beijing , China) Abstract: Objective To examine and identify fungi with important significance in food safety by matrix2assisted laser desorptionπionization time2of2flight mass spectrometry (MALDI2TOF2MS). Method Sample preparation, screening of appropriate matrix for specific fungal analysis, suitable ratio of matrix to sample, methods for loading samples on mass spectrometer sample tray and repeatability were investigated to optimize the parameters for fungal detection and identification by MALDI2TOF2MS. Results The results showed that different genus of fungi owned their proper matrix in sample preparation : MBTfor both Monascus and Fusaruim species, IAA for Aspergillus and Penicillium species as well as yeast, respectively. The optimum ratio of matrix to sample was 1 1 (vπv). Dried droplet is the best choice for applying sample on sample tray of the mass spectrometer. Conclusion MALDI2TOF2MS is of stability and repeatability characteristics in detecting and identifying fungus. Key word : Spectrometry, Mass, Matrix2Assisted Laser Desorption2Ionization ; Fungi ; Chemistry, Analytical, ( matrix2assisted,,,,,, : ( ) ; (2006BAK02A27) :, 20 80, laser desorptionπionization time of flight mass spectrometry, MALDI2TOF2MS),,,,,, [1-5 ] MALDI2TOF2MS

2 386 CHINESE JOURNAL OF FOOD HYGIENE ( Aspergillus species) ( A. flavus ) ,, , ( A. parasiticus) ,, ATCC2999 ( A. sojae) ( A. ochraceus ) ( A. oryzae ) ( A. niger) ( A. carbonarius) ( Penicillium species) ( P1 verrucosum ) ( P1 expansum) ( P1 urticae) 31833, ( P1 chysogenum) ( P1 purpurogenum) ( P1 islandicum) ( P1 rugulosum) ( Fusarium species) ( F1 graminaerum) ( F1 moniliforme) ( F1 tritinctum ) ( F1 proliferatum ) ( F1 oxysporum) ( F1 culmorum) ,7, ( Monascus species) ( Monascus ruber) SJS - 6 SJS - 8 SJS - 10 SJS - 18 SJS - 21 SJS - 22 SJS - 24 SJS - 26 SJS - 28 SJS - 33 ( Yeast) ( ), FX - 2X ( Saccharomyces cerevisiae) 1 2, ( Malt Extract Agar, MEA) - - ( Potato Dextrose Agar, PDA) (Malt Agar, MA) ( GBΠT ) [6 ] MALDI2TOF2MS 2, 5 - ( 2, 5 - dihydroxybenzoic acid, DHB) (2 - amino methyl nitropyridine, 2a4m5n) ( trans indoleacrylic acid, IAA) 2 - ( 2 - mercaptobenzothiazole, MBT) 2 - (4 - ) [ 2 - (4 - hydroxyphenylazo) benzoic acid, HABA ] 2, 6 - ( 2, 6 - dihydroxyacetophenone, DHAP) (Sinapinic acid, 3,5 - dimenthoxy hydroxycinnamic acid, SA) (- cyano hydroxycinnamic acid, CHCA,cyano) (5 - chloro mercaptobenzothiazole, CMBT), Sigma % ( Trifluoroacetic acid, TFA) ( , ) MALDI2TOF2MS 9, MALDI2TOF2MS ( Voyager DE TM PRO Biospectrometry TM Workstation),:,,, :20 kv, :400 ns, : Da, : 100, : 2010 Hz, : 9219 %, Bin size : 015 ns, Vertical scale : 500 mv, Vertical offset : 015 %, Input bandwith : 500 MHz, Guide wire :0105 %Angiotensin ( : ) ACTH (: ), : Da MALDI2TOF2MS SJS , MALDI2TOF2MS () SJS - 22 MEA,30 10 d ;

3 MA, PDA,28 7 d, 1 PDA,35 24 h ( ) 1 ml, 115 ml Eppendorf, rπmin 5 min, 2, 20 l 110 ml Eppendorf, SJS - 22 SJS - 22 ( , ),1 l MALDI2 TOF2MS MALDI2TOF2MS SJS l,,1 l, MALDI2TOF2MS, 1 l, MALDI2TOF2MS, 1 l,1 l, 1 l, MALDI2TOF2MS MALDI2TOF2MS SJS MEA, 7 d,maldi2 TOF2MS MALDI2TOF2MS MALDI2TOF2MS SJS MALDI2TOF2MS 1 ( mπz),1,, MBT, mπz 4 030,, MBT MALDI2 TOF2MS 212 SJS - 22 MBT ( ) MALDI2TOF2MS, 22, 1 + 1,, 1 + 1,, SJS ,,mΠz ;, ; mπz 4 030,,,MALDI2TOF2MS 214 MALDI2TOF2MS (1 + 1) 9 MALDI2TOF2MS, 4 56, IAA (mπz mπz mπz mπz ),IAA (mπz mπz mπz 9 326mΠz ),MBT (mπz mπz mπz mπz 1 759mΠz ),, MALDI2TOF2 MS,IAA MBT, IAA 215 MALDI2TOF2MS

4 388 CHINESE JOURNAL OF FOOD HYGIENE SJS SJS d MALDI2TOF2MS 7, 3 MALDI2TOF2MS,, mπz4 030,, MALDI2TOF2MS,

5 389 2 SJS - 22 MBT 3 SJS MALDI2TOF2MS,,,,, MALDI2TOF2MS,, [7 ] TFA, NH 2,,, TFA,,,,, [8 ] MALDI2TOF2

6 390 CHINESE JOURNAL OF FOOD HYGIENE MS : ; MALDI2TOF2MS,,MALDI2TOF2MS,,,

7 ,, MALDI2TOF2MS MALDI2TOF2MS [7 ], 9,,,, MALDI2TOF2 MS, [8 ],,,,

8 392 CHINESE JOURNAL OF FOOD HYGIENE ,,,,,, MALDI2TOF2MS

9 393 7 SJS - 22 MEA, , ( ), Π ( ),,,, ;, Π ( ) [1 ],,. [J ].,2004, 33 (5) : [2 ],. [J ].,2002,38 (4) : [3 ] WELHAM KJ,DOMIN M A,JOHNSON K, et al. Characterization of fungal sporesby laser desorptionπionazation time2of2flight mass spectrometry[j ]. Rapid Common Mass Spectrom,2000,14 : [4 ] NANCYB VALENTINE, JON H WAHL, MARK T KINGSLEY, et al. Direct surface analysis of fungal species by matrix2assisted laser desorptionπionazation mass spectrometry [ J ]. Spectrom,2002,16 : Rapid Common Mass [5 ] BIJAN AMIRI2ELIASI, CATHERINE FENSELAU. Characterization of Protein Biomarkers Desorbed by MALDI from Whole Fungal Cells[J ]. Anal Chem,2001, 73 (21) : [6 ] GBΠT [ S]. [7 ],,. Π [J ].,2004, 32 (12) : [8 ],,,. [J ].,2002, 30 (8) : [ : ] :R15 ;Q ;O65713 :A : (2007)

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