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High Performance Liquid Chromatograph (HPLC)                                                                              High Performance Liquid Chromatograph (HPLC)






            Tocopherols                                                                                                               Polycyclic Aromatic Hydrocarbons (PAHs)

            Tocopherols (Vitamin E) consist of a class of nutrients that are not only used as food additives such as antioxidants and nutritional   Polycyclic Aromatic Hydrocarbons (PAHs) are one group of wide-ranging environmental pollutants, some compounds of which have been
            supplements, but are also included in natural ingredients. Tocopherols are known to exist in various forms, including the β-, γ-, and   identified as being carcinogenic, toxic and mutagenic. The U.S. Environmental Protection Agency (EPA) has designated 16 polycyclic aromatic
            δ-isomers, not to mention α-tocopherol.                                                                                   hydrocarbons compounds as "priority pollutants" that are subject to restriction, and EPA methods 550.1 (drinking water) and 610 (waste
            HPLC analysis of tocopherol isomers is typically conducted using normal phase chromatography combined with fluorescence   water) specify test methods using HPLC and GC. Many polycyclic aromatic hydrocarbons are fluorescent, and can therefore be detected with
            detection. Shimadzu conducted ultra fast analysis using the Shim-pack XR-SIL high-speed, high-resolution analytical column (particle   high selectivity at high sensitivity using a fluorescence detector. Shimadzu analyzed a mixture of 15 PAHs, excluding acenaphthylene, which
            diameter 2.2 µm) together with the RF-20AXS detector. Below shows an example of analysis of a standard solution of 4 tocopherols   does not display fluorescence, using the RF-20AXS and time programmed excitation and emission wavelength switching.
            (2 mg/L each) using the Shim-pack XR-SIL column, which shortened the analysis time to less than one-fourth of the time when using a
            conventional column.
                                                                                                                                      Analytical Conditions
                                                                                                                                      Column       :SUPELCOSIL LC-PAH (50 mmL. × 4.6 mm I.D., 3 µm)
                                                                                                                                      Mobile phase   :A: Water   B: Acetonitrile
            Analytical Conditions                                                                                                     Time program   :B Conc. 50% (0-0.5 min)     88% (3.0 min)     88% (3.0-4.2 min)     100% (4.21-4.5 min)     50% (4.51-5.0 min)
            Column        :Shim-pack XR-SIL (75 mm L. × 3.0 mm I.D., 2.2 µm)                                                          Flow rate    :3.0 mL/min
            Mobile phase   :Hexane / 2-Propanol = 100 / 0.5 (v/v)                                                                     Column temperature :40°C
            Flow rate     :1.2 mL/min (XR-SIL), 1.0 mL/min (CLC-SIL(M))                                                               Injection volume   :5 µL
            Column temperature :30 °C                                                                                                 Cell temperature   :20ºC
            Injection volume   :4 µL (XR-SIL)                                                                                         Detection    :RF-20Axs
            Detection     :RF-20AXS Ex. at 298 nm, Em. at 325 nm                                                                                    0 - 1.54 min Ex. at 270 nm, Em. at 330 nm, Gain : × 1
            Cell temperature   :25 °C                                                                                                               1.54 - 1.89 min Ex. at 250 nm, Em. at 370 nm, Gain : × 1
            Flow cell     :Semi-micro cell (XR-SIL)                                                                                                 1.89 - 2.30 min Ex. at 330 nm, Em. at 430 nm, Gain : × 4
                                                                                                                                                    2.30 - 2.75 min Ex. at 270 nm, Em. at 390 nm, Gain : × 1
                                                                                                                                                    2.75 - 3.62 min Ex. at 290 nm, Em. at 430 nm, Gain : × 1
                                                                                                                                                    3.62 - 4.20 min Ex. at 370 nm, Em. at 460 nm, Gain : × 16
                                                                                                                                                    4.20 - 5.00 min Ex. at 270 nm, Em. at 330 nm, Gain : × 1






























                                         Chromatograph of a Standard Mixture of Tocopherols (2 mg/L each)





                                                                                                                                                                 Chromatogram of a Standard Mixture of Polycyclic Aromatic Hydrocarbons

                                                                                                                                      Reference: Application News L393 - Applications of the Prominence RF-20AXS Fluorescence Detector (Part 2)
            Reference: Application News No.L393 - Applications of the Prominence RF-20AXS Fluorescence Detector (Part 2) High-Speed Analysis of Tocopherols and Polycyclic Aromatic Hydrocarbons     High-Speed Analysis of Tocopherols and Polycyclic Aromatic Hydrocarbons

                                                                                                                                                                   Solutions for Palm Oil Analysis Application Notebook
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