Impurity Analysis Using a New High-Sensitivity UHPLC Detector

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Recently, a new high-sensitivity detector was introduced that provides improved UHPLC system performance and includes new features that improve workplace efficiency.

This presentation introduces this technology and provides a description of its use for impurity analysis. For more information, go to www.ssi.shimadzu.com or follow Shimadzu on Twitter @shimadzussi.

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Impurity Analysis Using a New High-Sensitivity UHPLC Detector

  1. 1. 1 / 13 Impurity Analysis Using a New High-Sensitivity UHPLC Detector William Hedgepeth, Kenichiro Tanaka Shimadzu Scientific Instruments, Columbia, Md., USA
  2. 2. 2 / 132 / 13 Introduction The use of UHPLC continues to grow throughout the analytical industry as researchers increase their need for faster LC separations with more sensitive detection requirements. Recently, a new high-sensitivity detector was introduced that provides improved UHPLC system performance and includes new features that improve workplace efficiency. The new UHPLC detector features a total reflection capillary cell to reduce system dispersion and allow ultra-high speed analysis. The high sensitivity of the detector combined with its wide dynamic detection range allows simultaneous analysis of a main peak and an impurity peak with a 0.005% concentration. A new function of the detector provides extended dynamic range (i-DReC). This function enables an extension in apparent dynamic range by shifting the spectrum wavelength when the linearity of the calibration curve cannot be obtained due to intensity saturation of the detector. Potential analyst error is also avoided in the dilution procedure. Results presented will demonstrate the increased sensitivity of the new PDA detector. An example of the detector’s extended dynamic range will also be presented.
  3. 3. 3 / 133 / 13 Needs for High-Sensitivity Detection Pharmaceutical • Growing interest in impurity analysis in genotoxicity test • Publication of the related guidance by FDA ⇒ Requires analysis of trace impurities (equivalent to 0.005%) Chemicals and Cosmetics • Increasing demand for a PDA detector for the quantitation and qualitative analysis of quasi- drugs • Simultaneous confirmation and qualitative test by retention time and spectrum Food Inspection Agency • Necessity for analyses of impurities, toxic substances, and foreign substances with increasing awareness of food safety • Increasing analysis of harmful components such as pesticides and melamine in food due to globalization of food distribution
  4. 4. 4 / 134 / 13 High-Sensitivity Cell Figure 1: 85 mm High-Sensitivity Flow Cell Standard cell: 10 mm optical length High-sensitivity cell: 85 mm optical length The Cell ID information can be shown with analytical results in the report.
  5. 5. 5 / 135 / 13 Sensitivity Comparison Enhanced signal by optional high-sensitivity cell ⇒ 85 mm optical length increases signal significantly
  6. 6. 6 / 136 / 13 Analysis of Ketoprofen System: Nexera SR (CBM-20A, DGU-20A5, LC-30AD x 2, SIL-30AC, CTO-30A, SPD-M30A) Column: Shim-pack XR-ODSIII 150x2.0mm Mobile phase: A: 0.1% Formic acid in water, B: Acetonitrile B.Conc 30%-70% (10min) Flow rate: 0.5mL/min Mixer: 180uL II Column temp.: 40°C Injection volume: 2uL Detection: 254 nm Cell: High-sensitivity cell (85mm) Sample: Ketoprofen
  7. 7. 7 / 137 / 13 Analysis of Ketoprofen 4.50 4.75 5.00 5.25 5.50 5.75 min 50 100 150 200 250 300 350 400 450 500 550 600 650 700 750 uV Data2:9122013_Ketoprof en_0.02mg$L_30%-70%(10min)m20a_002.lcd PDA Ch1 254nm,4nm Data1:9182013_Ketoprof en_0.02mg$L_30%-70%(10min)_002.lcd PDA Ch1 254nm,4nm Figure 2: Chromatogram of Ketoprofen showing high sensitivity vs. standard cell
  8. 8. 8 / 138 / 13 Trace Level Impurity Detection Figure 3: Peaks with areas of less than 0.0001% could be detected Datafile Name:9162013_Ketoprofen_500mg$L_30%-70%(10min)rinse_039.lcd Sample Name:Ketoprofen Sample ID:500mg/L 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 min -1 0 1 2 3 4 5 6 mAU 254nm,4nm Ketoprofen Datafile Name:9162013_Ketoprofen_500mg$L_30%-70%(10min)rinse_039.lcd Sample Name:Ketoprofen Sample ID:500mg/L 3.25 3.50 3.75 4.00 4.25 4.50 4.75 5.00 5.25 5.50 5.75 6.00 6.25 min 0.0 0.1 0.2 0.3 0.4 0.5 0.6 mAU 254nm,4nm /0.019805 /0.000133 /0.000031 /0.001622 /0.000266 /0.000055 /0.000077 Ketoprofen/99.900219 /0.003438 /0.012885 Data File Name: 9162013_Ketoprofen_500mg$L_30%-70% (10 min) rinse_039.lcd Sample Name: Ketoprofen Sample ID: 500 mg/L
  9. 9. 9 / 139 / 13 Carryover Analysis 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 min -500 0 500 1000 1500 2000 2500 3000 3500 4000 4500 uV Data3:9122013_Blank__30%-70%(10min)rinse_007.lcd PDA Ch1 254nm,4nm Data2:9132013_Ketoprofen_500mg$L_30%-70%(10min)rinse_020.lcd PDA Ch1 254nm,4nm Data1:9132013_Ketoprofen_0.01mg$L_30%-70%(10min)30min_002.lcd PDA Ch1 254nm,4nm Figure 4: Ketoprofen chromatograms showing 0.01 ppm, 500 ppm, and post-run blank (top to bottom)
  10. 10. 10 / 1310 / 13 Extended Linearity The newly developed intelligent Dynamic Range Extension Calculator (i-DReC) function can expand the dynamic range in the high-concentration region. It provides a powerful tool for checking synthetic compounds, stability testing, and the simultaneous qualification of principal components and trace impurities. The calibration curve linearity may become lost due to signal saturation if the absorbance is extremely high, when analyzing high-concentration samples, for example. The LabSolutions i-DReC function corrects the linearity in the high-concentration region by shifting the spectrum wavelengths and performing correction calculations using signals at wavelengths where linearity is achieved. The i-DReC function can improve the linearity of the absorbance range over 4 AU to permit analysis of higher concentration samples.
  11. 11. 11 / 1311 / 13 Extended Linearity Using the high-sensitivity cell and the i-DReC function can further extend the applicable Nexera X2 sample concentration range. The wider concentration range suits not only checking synthetic compounds and stability testing but also trace impurity analysis, such as the genotoxicity testing of drugs. Labor to change dilution ratios of each compound is reduced and the risk of human error is avoided.
  12. 12. 12 / 1312 / 13 Extended Linearity
  13. 13. 13 / 1313 / 13 Summary A new UHPLC detector was introduced that allows low-level impurity analysis down to 0.0001%. The optional 85 mm high-sensitivity cell provides the highest sensitivity available in PDA detectors while keeping peak dispersion to a minimum.
  14. 14. 14 / 13 Thank you for viewing this presentation. Should you have any questions or require additional information about our research, products or services, please visit our support page: www.ssi.shimadzu.com/support/ @shimadzussiFollow us on Twitter Need More Info?

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