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Analysis of Mycotoxins  Identification and Observation of Microorganisms






 Mycotoxins are chemical products produced in fungus and are toxic to humans, animals, and crops. There are more than 300   In the event of food poisoning, identification of the causative factor is extremely important in determining the path of
 known mycotoxins, including aflatoxin, deoxynivalenol, patulin, fumonisin and others, and some of them are strictly   infection, prescribing treatment and preventing recurrence. Shimadzu’s iDPlus microorganism identification platform provides
 controlled under regulations. HPLC and LC-MS are mainly used for analysis of mycotoxins, and simultaneous analysis of   rapid identification of microorganisms in food.
 multiple mycotoxins has been carried out for more efficient screening.
                    Identification of Microorganisms (MALDI-TOF-MS)     AXIMA Microorganism Identification System
                    In order to prevent pathogenic germs from contaminating
 Analysis of Aflatoxins (LC)  Rapid Screening of 10 Mycotoxins (LC)
                    food sources, it is essential to identify them quickly and
 The analysis of aflatoxins (B1, B2, G1 and G2) is very   Simultaneous analysis of mycotoxins is important because   reliably in order to allow for proper controlled measures.
 important from a food safety perspective due to concerns   multiple mycotoxins have been detected in a single food and   A microorganism identification system using the AXIMA
 about adverse effects on human health. The Nexera Ultra   may cause a synergistic effect. From this background, a   MALDI-TOF MS enables direct measurement of cultured
 High Performance Liquid Chromatograph achieves   simultaneous analytical method has been developed for   microorganisms and identification within two minutes.
 high-resolution analysis of aflatoxins in less than four   rapid screening of 10 mycotoxins using HPLC. Shown below
                      % Int.
 minutes. The RF-20Axs high-sensitivity fluorescence detector   are chromatograms of 10 mycotoxins by fluorescence (upper)    Salmonella enterica  subsp.  Enterica
 allows direct analysis of aflatoxins at trace levels (ppt)   and PDA (lower) detectors without derivatization.  4365.39   5369.13   6091.59
 without any derivatization.                    10950.18
 mV                                7158.16   7658.66
 3                     3829.04
 Integrated HPLC  RF-20AXS  4  ■Peaks      9520.27
     Peaks  Nexera-i  2  1) AFM1 1 µg/L  6885.93
 2) AFG2 0.25 µg/L
 1 AFG2 (0.6 µg/kg)  2  3) AFG1 0.25 µg/L
 2 AFG1 (2.4 µg/kg)  1  4) AFB2 0.25 µg/L
 3 AFB2 (0.6 µg/kg)  5) AFB1 0.25 µg/L  5033.17   5524.87    Staphylococcus aureus  subsp.  Aureus
 4 AFB1 (2.4 µg/kg)  6) ZON  50 µg/L  4308.68
 1  7) OTA  5 µg/L              6352.79
 7                                                   12397.15   13762.64
 5                     3640.52
 0  3  6
 4  5  6  7  8  9  10  11  4434.57   5209.27   6043.95   6671.91   7230.02   8348.89    Pseudomonas aeruginosa
 min                                 7886.39     11424.07
 mAU                    4054.41     7590.49   10220.70   11027.94   11922.76   12295.04   12706.04   13225.69   13698.41   14067.89   14991.82
 Nexera-i PDA  ■Peaks
 5  8) NIV  1000 µg/L
 8  9) DON 1000 µg/L               7156.44
 10) PAT 25 µg/L                6253.14   9059.12   9733.78    Escherichia coli
 9                                   7867.55                           Features
 Analysis of Aflatoxins without TFA Derivatization (N = 5)  8364.74
 0  220 nm                               8871.39
 10                                                      13645.06      Quick and Easy Analysis
 Source: Shimadzu Application News No. L435  276 nm
 0.0  0.5  1.0  1.5  2.0  2.5  3.0                       Bacillus  subtilis      Three steps from sample preparation to identification.
 min                     4307.86   5255.03   9235.80                    The microorganism identification process is finished within two
 Simultaneous Analysis of 10 Mycotoxins  3890.60
 Simultaneous Analysis of 25 Mycotoxins (LC-MS/MS)  Source: Shimadzu Application News No. L512  minutes after starting the analysis.
                       4000   6000   8000   10000   12000   14000      Highly Accurate Identification
 LC-MS/MS is effective for analysis of mycotoxins in complex   m/z      Microorganism identification by MALDI-TOF MS is a well-established
 matrices. Shown is an example of the simultaneous analysis             technique with a low number of false positives.
                              Comparing mass spectra of different bacteria
 of 25 mycotoxins by LC-MS/MS and the Method Package for   Database  Source: https://www.shimadzu.com/an/industry/pharmaceuticallifescience/proteome0207005.htm  Applicable for a Wide Range of Microorganisms
 Mycotoxins. All compounds, including positive and negative             Capable of classifying/identifying gram-positive and gram-negative
 ions, were separated and detected with high sensitivity   LC/MS/MS Method Package for Mycotoxins  What is Microorganism Identification by MALDI-TOF MS?  bacteria, yeasts, fungi and spores. iDPlus identifies mircoorganisms
 The Method Package provides simultaneous multi-component analysis      by family, genus, species, and subspecies.
 within 12 minutes. For analysis of mycotoxins with high
 conditions for mycotoxins subject to regulations in the US, the EU,   Species-specific mass spectral patterns from microorganisms can be   Database Customization
 sensitivity, eliminating carryover is indispensable. Modified
 China and Japan. It contains seven different analytical method   obtained by direct measurement of microorganisms using MALDI-TOF   Easily register new microorganisms in the database, which allows
 rinse methods using the SIL-30AC autosampler can eliminate
 conditions and fully optimized MRM transitions for a total of 27   MS. Identification of microorganisms can be performed by matching   rapid development of a high-quality database.
 carryover of fumonisins.
 different mycotoxins. Rapid analytical methods are included for a 25   these collected spectra to the database. This is a fast, easy and   Low Operational Cost
 mycotoxin screen, 16 mycotoxins using high-sensitivity conditions, 10   reliable method for identification of microorganisms.  Analysis can be performed for pennies per sample compared to
 mycotoxins using APCI, and specialized methods for trichothecenes,     expensive traditional techniques.
 fumonisins, aflatoxins, and patulin.                                  Expanded Research Capabilities
                                                                        iDPlus is not limited to microorganism identification.  Reflectron
 Number of  Analysis                                                    mode and MS/MS functionalities enable a true research platform.
 Method
 Components  Time (min)
 Simultaneous Analysis of 25 Mycotoxins  25  15
 Simultaneous Analysis with High Sensitivity  16  30
 Patulin Analysis  1  10
 Aflatoxins Analysis  6  30
 Trichothecenes Analysis  4  12
 High-speed Simultaneous Analysis of 25 Mycotoxins  Fumonisins Analysis  3  15
 Source: Shimadzu Flyer No. C146-E351  10 mycotoxins using APCI  10  33
                                                                 Shimadzu’s Total Support for Food Safety
 8                                                                                                Analytical and Testing Instruments for Food Safety  9
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