Page 12 - Shimadzu Journal vol.2 Issue1
P. 12

One of the things we do is use LC for quantitative analysis in
                                                                                                                                      early-stage drug discovery work. Unlike many physics, chemistry, and
                                                                                                                                      drug-manufacturing departments, where analysis is performed using
                                                                                                                                      validated instruments or measurements are made for application
                                                                                                                                      processes, we focus not only on data quality and accuracy, but also on
                                                                                                                                      analytical efficiency to process as many compounds as quickly as possible.
                                                                                                                                      The samples Shimadzu tested for us contained high concentrations of
                                                                                                                                      compounds that would normally be diluted before quantitation.
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            Pretreatment processes such as dilution can affect accuracy. In
            addition, dilution and some other pretreatment processes require                                                            Great Potential for Further Applications
            considering the solvent used, which takes additional time. Highly viscous
            solvents, moreover, require separate consideration. For example, the                                                      High-Sensitivity Cell and New Dynamic Range Extension   Features:
            sample quantity available for administration tests is often very limited, so                                              Function Extend Measurement Concentration Range of
            we were already looking for ways to eliminate such negative factors by                                                    Nexera X2 Series UHPLC by a Factor of 50           •  The i-DReC function can be used with either SR-Cells (10 mm optical
            measuring concentrated samples directly without pretreatment.                                                             LC analysis often requires performing multiple analyses to analyze   path length) or high-sensitivity SR-Cells (85 mm optical path length)
            However, typical PDA detectors have a limited dynamic range.                                                                                                                 and measurements can be performed using a single PDA detector.
                                                                                                                                      samples with different concentration levels. For instance, when using
            Eventually, we considered using MS or another detection method with a                                                     the same detection method to quantify compounds with significantly   •  Using a high-sensitivity SR-Cell boosts the sensitivity of SPD-M30A
            wide dynamic range, but MS requires dealing with matrix effects and                                                       different concentration or sensitivity levels, or when measuring   detectors by about 5 times and allows detecting extremely small
            considering internal standard correction. It was at that point that we                                                    impurities, other trace components, and primary components.  peaks.
            were introduced to the i-DReC and provided samples. I thought the                                                         Nexera X2 systems, when equipped with an SPD-M30A   •  The i-DReC function extends the measurement range by about 10
            technology was theoretically possible, but after seeing the good range                                                    high-sensitivity cell (85 mm optical path length) and with the i-DReC*   times in the high-concentration direction and automatically corrects
            and linearity of calibration curve results, I realized that this was truly a                                              function enabled, minimize this issue. With such a configuration,   the peak area and height values of saturated peaks.
            useful function.
                                                               principal components are often detected at their optimal wavelengths,   Nexera X2 systems can obtain information about both extremely small
                                                               which are not necessarily the same as the optimal UV wavelengths for   and large peaks from a single analysis. Consequently, Nexera X2   •  Using the i-DReC function with the high-sensitivity SR-Cell extends
            Did you have any apprehensions about the i-DReC    impurities or raw material components. Consequently, detection can     systems can reduce the number of analyses required and increase   the Nexera X2 measurable concentration range by up to 50 times.
            technique?                                         require considerable effort. Also, if only MS is used for analysis, the   productivity. In particular, it provides an effective way of shortening   •  Obtain results from a single dataset acquired using a single PDA
                                                               raw materials or other components can be made up of                    the time (and improving efficiency) of analyses when each   detector.
            I knew it was important to specify optimal wavelength settings, but I   low-molecular-weight molecules, which can prevent adequate   measurement takes a long time, due to various constraints on
            didn't know what would result from extremely low molecular weight   detection and make it difficult to determine appropriate analytical   analytical conditions, for example.  •  i-DReC is included as a standard feature of LabSolutions software.
            causing a lack of multiple maximum absorption wavelengths, from   conditions for simultaneous quantitative analysis. Similarly, when
            barely detectable peaks on the low wavelength end, or from very low   considering processes, reactions must be checked with simultaneous
            absorptivity. As it turned out, when we calculated the linearity for even   quantitation that is conducted in as short a time as possible. In
            rather low-wavelength and sloped regions of the spectrum, linearity   contrast, if calibration curves are prepared in advance, quantitative
            was quite high, which convinced us that it was a useful technique.  analysis can be accomplished very quickly by loading PDA data and
                                                               using the i-DReC function.
            What about in terms of improving analytical efficiency?

            It definitely allows us to improve analytical efficiency. With   Did you see the i-DReC settings screen in LabSolutions?
            conventional techniques, dilution requires particular care, because not
            only the type of solvent, but also the solubility is important. If viscosity   Yes, I did. Only two windows were added and there were few settings
            is too high, it can also affect results from LC analysis. It is also   needed to be specified. I think it only involved setting wavelength and
            important to determine a dilution rate that allows the concentration   threshold values. I was shown on the spot how to generate results,   This ability to measure a wider range of concentrations means the   Using the i-DReC function in combination with a high-sensitivity
            of a target component to fall within the quantitative range of the   which did not seem very difficult. Without having to specify all sorts of   system can now be used for an even wider range of applications. These   SR-Cell requires only a single analysis of the sample to detect peaks (1)
            calibration curve. Furthermore, since calibration curves are often   parameters, it seemed rather user-friendly.          include analyzing ultra-trace impurities, trace contaminants or   through (8). Then, based on those impurities, area value results can be
            prepared using standard samples, subtracting the background noise                                                         hazardous substances in genotoxicity testing in the pharmaceutical   obtained for primary components by using pre-specified threshold
            from the diluting solvent must be considered. Background noise                                                            industry, and confirming the purity, monitoring the processing, or   values to automatically correct area values as though the peaks had
            effects can be quite large in MS, which is why an internal standard   What do you think about the reliability of results   testing the stability or dissolution of synthetic compounds.  not been saturated. In this example, results with an extremely small
            needs to be added. Using MS provides a wide dynamic range, but it   obtained using i-PDeA?                                                                                   error factor were obtained from analyzing two sample concentrations.
            requires considering a lot of factors. Therefore, I think that in reality,                                                Furthermore, results can be obtained from a single dataset acquired
            PDA results provide higher accuracy for our measurement purposes.  When it comes to data reliability, I am concerned about how the data   using a single PDA detector. This makes it easier to obtain more   In contrast, using previous methods, peak information for primary
                                                               may be interpreted. After all, the information in the final results is not   consistent results than when changing the concentration level and   components was first obtained by analyzing a low-concentration
                                                               extracted directly from raw data. On the other hand, it is not that data   performing multiple analyses. The following is an example of results   sample (50 ppm). Then the peak information for trace components
            i-DReC is only one of the PDA functions, so have you   taken from different instruments are combined into one, but rather   obtained from analyzing a pharmaceutical mixture sample using a   was obtained by analyzing a high-concentration sample (1000 ppm).
            thought of connecting the PDA detector in series with an   that information from simultaneously acquired data is used. Therefore,   Nexera SR system (with a high-sensitivity SR-Cell).  This required calculating the total area ratio percent value.
            MS system after it in some cases?                  it may be safe to conclude that no analytical problems arise as long as
                                                               data consistency is maintained. I think it would be important to        Table 1: Comparison of Area% Corrected by i-DReC and Obtained from

            Yes, it would probably be used more often as an LC/MS system,                                                              ɹɹɹɹAnalyzing Two Sample Concentrations
            because we also want to obtain MS data.            confirm consistency by using actual samples to compare results from                                  Area Ratio (%)       * i-DReC is a function that determines
                                                               previous methods with results obtained using i-PDeA. In that respect,            Retention Time  Area Ratio (%)  Obtained from  peak area and peak height in PDA
                                                               the tests conducted by Shimadzu probably serve to demonstrate             Peak No.  (min)  Corrected by  Two Samples  Error  detector data with peak intensity
                                                                                                                                                                    (50 ppm and
            Aside from quantitation of high-concentration samples,   satisfactory data consistency for the given samples.                                   i-DReC   1000 ppm)           saturation (intensity has exceeded a
            would you use it for simultaneous quantitation of                                                                          ɹɹɹ(1) Main  4.634   96.6789   96.7171   -0.0382  pre-specified threshold value) by
            components with large differences in concentration,   Thank you so much for providing your samples and                       (2)       5.448    2.8749    2.8419    0.0330   obtaining a chromatogram that is
            such as impurities and principal components?       helping us with your valuable comments.                                   (3)       3.900    0.1993    0.1970    0.0023   automatically shifted to a position
                                                                                                                                         (4)
                                                                                                                                                                      0.1007
                                                                                                                                                            0.1019
                                                                                                                                                   4.910
                                                                                                                                                                                0.0012
                                                                                                                                         (5)       5.091    0.0469    0.0464    0.0005   where peaks are at a wavelength with
            When we study the synthesis process for manufacturing, we must                                                               (6)       4.487    0.0295    0.0291    0.0004   low absorption. Then, the sensitivity
            consider the synthesis route. Therefore, it is important to establish                                                        (7)       4.226    0.0248    0.0245    0.0003   ratio between the wavelengths in the
            analytical methods (simultaneous multi-component analysis) that can                                                          (8)       4.975    0.0248    0.0245    0.0003
                                                                                                                                         Imp1      4.056    0.0062    0.0061    0.0001   spectral data is corrected to calculate
            quantitatively determine, based on how long the reaction is
                                                                                                                                         Imp2      4.331    0.0076    0.0075    0.0001   the peak area and height values at the
            performed, the amount of principal components produced and the                                                               Imp3      4.376    0.0052    0.0051    0.0001   target wavelength.
            loss of impurities and raw materials, for example. In such cases, the
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