Page 8 - Microorganism Species Analysis

LCMS-2020 LCMS-2020
Low-Molecular Information
Microorganism Solutions Microorganism Solutions

Microorganism Classification by Chemical Analysis Data
In addition to gene analysis or cultivation, staining, and shape observation, microorganisms can also be classified by the
chemical analysis of the microorganism components. Methods previously known for the classification of microorganisms Analysis of the Actinomycete Menaquinone Using LC/MS
include using the guanine-cytosine content of the constituent DNA as an index for the microorganism genome and the GC or
The analysis of the bacterial fatty acid composition and isoprenoid quinones has conventionally been used as a Microorganism Species Observation of
GC/MS analysis of the fatty acid cell membrane components after extraction and derivatization.
chemical classification indicator for actinomycetes.
LC/MS is a powerful tool for the analysis and taxonomic evaluation of compounds such as isoprenoid quinones with multiple
Typical menaquinone molecule species have been widely researched and provide an extremely effective method for
similar structures.
Observation of
classification and identification at the genus level. This composition nomenclature is essential for naming a new
species. LC/MS is a powerful tool for the identification and determination of the composition ratio of menaquinone
Microorganism Species
molecule species.

Features of the LCMS-2020 Ultra Fast Liquid Chromatograph Mass Spectrometer MK-n (Hm) n: number of isoprene units
············· Expression (1) m: number of hydrogen atoms related to saturation of the isoprene side chains
Rapid 15 msec positive/negative ion switching time permits high-speed detection (MS measurements) that follows
high-speed LC separation. Microorganism Species Identification of
The newly developed Qarray ion optical system achieves superior sensitivity, reproducibility, and linearity. Fig. 1 Chemical Structural Formula of Menaquinone
Rapid, 15,000 u/sec scanning achieves high ion permeability while maintaining resolution.
Identification of
The LCMS-2020 dramatically enhances long-term stability and ease of maintenance. Test and research flow
LCMSsolution maximizes analysis performance through comprehensive functions including data comparisons, peak
Microorganism Species
1. Pulverize freeze-dried bacteria (100 to 300 mg) and extract the lipid components using chloroform/methanol (2:1).
integration, and report output.
2. Re-dissolve the filtrate in acetone, and roughly separate the menaquinones using TLC silica gel.
3. Re-extract and re-concentrate the menaquinones and perform LC/MS analysis (in ESI-positive mode).

Analytical Conditions
Int. TIC Column : Shim-pack FC-ODS
717.00
719.00 Mobile phase : acetonitrile / methanol /
721.00 MK-8(H8) 10 mM ammonium acetate = 2 / 6 / 15
2000e3 723.00 MK-8(H6) Specific Microorganisms Detection of
725.00 Flow rate : 0.2 mL/min
727.00
729.00 Column temp. : 40 °C
1500e3 731.00
733.00 Injection volume : 6 µL
735.00 Probe voltage : 4.5 kV (ESI-positive mode)
Detection of
1000e3
CDL temp. : 300 °C
500e3 Nebulizing gas flow : 1.5 L/min
Drying gas pressure : 0.15 MPa
Specific Microorganisms
0e3 CDL voltage : 25 V
5.0 10.0 15.0 20.0 25.0 30.0 35.0 min Qarray DC voltage : Scan mode
Int. TIC
785.00 Qarray RF voltage : Scan mode
787.00 MK-9(H8)
789.00 MK-9(H6) Scan range : m/z 200-400 (1.0 sec/scan)
791.00
7.5e6 793.00 SIM : m/z 331.1, 329.1, 315.1, 313.0 (0.25 sec/ch)
795.00 MK-9(H4) MK-9(H10)
797.00
799.00 MK-9(H12)
781.00
5.0e6 783.00
Specific menaquinone molecule species are
2.5e6
detected for each actinomycete (Fig. 2 and Table 1).
LCMS-2020 0.0e6 5.0 10.0 15.0 20.0 25.0 30.0 35.0 min Analysis based on such a chemical classification Components Analysis of Microorganism-Derived
Ultra Fast Liquid Chromatograph Mass Spectrometer index is important for the classification and
Fig. 2 Total Ion Chromatogram and Mass Chromatogram of Menaquinones identification of bacteria. LC/MS is an essential
Components
analysis tool for this purpose.
Analysis of Microorganism-Derived
Table 1 Menaquinone Molecule Species Content and Relative Retention Time based on MK-6 (H0) for Typical Actinomycete Species
Actinomycetes MK-7 MK-8 MK-9 MK-10
JCM No. H0 H0 H2 H0 H2 H4 H6 H8 H0 H2 H4 H6
Thermoactinomyces candidus 3180T 1.00 1.50
Rhodococcus equi 1311T 1.77
Amycolata autotrpphica 4348T 2.05
Nocardia asteroides 6043T 1.86 Other Applications
Tsukuamurella pauromethabolum 3226T 1.87 2.11 2.38
Streptosporangium vulgare 3028T 2.47
Amycolatopsis orientalis sub sp. Orientalis 4600T 2.39 2.70 3.07
Actinomadura madurea 7436T 2.66 3.04
Other Applications
Browser screen Analytical conditions optimization screens Streptomyces albus subsp. Albus 4450T 2.67 3.05
Kitasatosporia setae 3304T 2.53 2.91 3.36 3.81
LCMSsolution Ver.5 Nocardiopsis assonvillei subsp. Dassonvillei 7437T 2.39 3.32
LCMS-2020 Workstation Saccharothrix australiensis 3370T
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