Page 25 - Microorganism Species Analysis
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Prominence UFLC                                                                                                                                                                                                      Prominence UFLC
                                            Analysis of Amino Acids, Organic Acids, and Aflatoxins
            Microorganism Solutions                                                                                                                                                                                              Microorganism Solutions




                     UFLC Analysis of Microorganism-Derived Components                                                                    Data
                     High-performance liquid chromatography is widely used for the analysis of amino acids in fermented and brewed food
                     products. However, the demand for fast analysis has been increasing over recent years to enhance the productivity of   Fast Analysis of Amino Acids by Pre-Column Derivatization
                     laboratory analysis. The Prominence Ultra Fast Liquid Chromatograph (UFLC) is based on a comprehensive analysis of
                                                                                                                                                                                       Derivatization Reaction with Phenylisothiocyanate (PITC)        Microorganism Species  Observation of
                     technologies to increase the speed of liquid chromatography. In addition to ultra-fast liquid chromatography, it achieves high   Standard sample
                                                                                                                                                                                       1. To 200 µL sample, add 100 mmol/L phenylisothiocyanate
                     levels of accuracy, stability, durability, and expandability not available from conventional HPLC. The ultra-fast analysis and
                                                                                                                                                                                           (acetonitrile solution) and 1 mol/L triethylamine (acetonitrile
                     high accuracy and reliability offered by the Prominence UFLC achieve revolutionary improvements in laboratory analysis
            Observation of
                                                                                                                                                                                           solution), agitate, and leave at 40 °C for 20 minutes to react.
                     productivity.
                                                                                                                                                                                       2. Allow to cool to room temperature, and spike with 1.2 mol/L
          Microorganism Species
                                                                                                                                              Commercial soy sauce
                                                                                                                                                                                           hydrochloric acid aqueous solution to neutralize.
                                                                                                                                                                                       This is an example of the fast analysis of amino acids by
                        Features of the Prominence UFLC Ultra Fast Liquid Chromatograph
                                                                                                                                                                                       pre-column derivatization using PITC. Analysis was performed on
                         Ultra-fast LC analysis: just one-tenth the analysis time of general LC                                                                                        amino acids, 23 related components (0.5 µmol/L), and commercially
                         Shorter total analysis cycle time                                                                                                                             available soy sauce. The analysis was completed within 10
                         High-resolution analysis: three times the resolution of general LC                                                                                            minutes.
                                                                                                                                           Analytical Conditions                                                                                       Microorganism Species  Identification of
                         High accuracy, stability, and durability                                                                          Column     : Shim-pack XR-ODS (100 mm L. × 3.0 mm I.D.)
                                                                                                                                           Mobile phase  : A) 10 mmol/L (potassium) phosphate buffer (pH7.0)
                         The Shim-pack XR-ODS Column for fast analysis and high separation maximizes the Prominence UFLC                                B) Acetonitrile
                         performance.                                                                                                                   B.conc; 5% (0 to 0.5 min), 5% to 35% (0.5 to 10.5 min)
                                                                                                                                           Flow rate  : 0.9 mL/min
            Identification of
                                                                                                                                           Column temp.  : 40 °C
                                                                                                                                           Detection  : absorbance at 254 nm with semi-micro flow cell
          Microorganism Species
                                                                                                                                           Sample volume  : 1 µL
                                                                                                                                           Fast Analysis of Organic Acids

                                                                                                                                             mAU                                         The ion exclusion mode is often used for the analysis of organic
                                                                                                                                            8                1                           acids, but in this case an ODS column that can handle 100%
                                                                                                                                                                 2           4
                                                                                                                                            7                                                                                                          Specific Microorganisms  Detection of
                                                                                                                                                                                         aqueous solutions was used to complete fast analysis within two
                                                                                                                                            6
                                                                                                                                                                                         minutes.
                                                                                                                                            5                        3
                                                                                                                                            4
                                                                                                                                                                                         Analytical Conditions
            Detection of
                                                                                                                                            3
                                                                                                                                                                                         Column    : Phenomenex Synergi 2.5 µm Hydro-PP 100A (100 mm L. × 3.0 mm I.D., 2.5 µm)
                                                                                                                                            2                                            Mobile phase  : 10 mmol/L (sodium) phosphate buffer (pH2.6)
                                                                                                                                            1                                            Flow rate  : 0.8 mL/min
          Specific Microorganisms
                                                                                                                                            0                                            Column temp.  : 30 °C
                                                                                                                                                                                         Injection volume : 4 µL
                                                                                                                                            0.00  0.25  0.50  0.75  1.00  1.25  1.50  1.75  min
                                                                                                                                                                                         Detection  : SPD-20A at 210 nm
                                                                                                                                                                                         Flow cell  : semi-micro cell
                                                                                                                                           Fast Analysis of Cephem Antibiotics
                                                                                                                                           Analytical Conditions
                                                                Prominence UFLC                                                            Column  : Shim-pack XR-ODS                                                                                  Components  Analysis of Microorganism-Derived
                                                                                                                                                     (100 mm L. × 3.0 mm I.D., 2.2 µm)
                                                                                                                                                     Shim-pack VP-ODS
                                                                                                                                                     (250 mm L. × 4.6 mm I.D., 4.6 µm)
          Components
                                                                                                                                           Mobile phase  : A:0.1% formic acid-water
                                                                                                                                                     B:acetonitrile
                                                                                                                                           Time program  [XR-ODS]
                                                                                                                                                     B.conc; 15% (0 min)
                                                                                                                                                     55% (3.5 min)
                                                                                                                                                     15% (3.51-6.5 min)
            Analysis of Microorganism-Derived
                                                                                                                                                   [VP-ODS]                                                 Structure of Cephem Antibiotics (Partial)
                                                                                                                                                     B.conc; 15% (0 min)
                                                                                                                                                     55% (30 min)
                                                                                                                                                     15% (30.01-45 min); 0.2 mL/min                      Cephem antibiotics are a type of beta-lactam
                                                                                                                                           Flow rate  : 1.0 mL/min (XR-ODS)                              antibiotic. They are broad-spectrum
                                                                                                                                                     1.0 mL/min (VP-ODS)
                                                                                                                                           Column temp.  : 40 °C                                         antibiotics with a strong antibiotic action, and
                                                                                                                                           Injection volume: 4 µL (XR-ODS)
                                                                                                                                                    10 µL (VP-ODS)                                       are used as general-purpose oral or injected
                                                                                                                                           Detection  : photodiode array                                 antibiotics. The combination of Prominence    Other Applications
                                                                                                                                                     UV-VIS detector 260 nm
                                                                                                                                           Flow cell  : semi-micro cell (XR-ODS)                         UFLC and Shim-pack XR-ODS increases the
                                                                                                                                                     conventional (VP-ODS)
                                                                                                                                                                                                         mobile phase linear velocity to approximately
                                                                                                                                                                                                         2.4 times that of the Shim-pack VP-ODS
          Other Applications
                                                                                                                                                                                                         during analysis. In addition, it maintains high
                                        LCsolution Screen                        Shim-pack XR-ODS                                                                     Chromatogram of 13 Cephem Antibiotic Components   resolution while reducing the analysis cycle
                                                                                 Column for Fast Analysis and High Separation                                              (Upper: Shim-pack VP-ODS;     time by approximately 85%.
                                                                                                                                                                            Lower: Shim-pack XR-ODS)
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