Page 12 - Shimadzu Journal vol.2 Issue1

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.
Customer Interviews Product Information
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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