by Nexera-i and LCMS-2020 No - Shimadzu · 2018. 11. 1. · LCMS-2020. The built-in UV/ VIS...
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Application News No. L465 High Performance Liquid Chromatography Investigation of Synthetic Compounds in Drug Discovery by Nexera-i and LCMS-2020 LAAN-A-LC-E242 n Investigation of Synthetic Compounds in Open Access Environment A workflow in which multiple analysts in an open access environment can quickly verify the synthesis of a variety of compounds using the same method is perceived as one of the solutions that can lead to greater efficiency in the research and development process. This Application News describes an example in which measurement was conducted for a range of compounds with properties spanning acidic to basic, and high to low polarity, all using the same analytical conditions (Table 1). Each of the various compounds was dissolved in dimethylsulfoxide (DMSO), and 0.1 µL was injected after diluting with methanol solution containing 5 % DMSO (Final concentration 20 mmol/L each). MS analysis was conducted using scan mode (m/z = 165 – 385) and positive / negative mode. The composition analysis results for the eight pharmaceutical compounds are shown below (Fig. 1.1 – 1.4). Also, assuming an actual workflow, we injected a blank solution (methanol) prior to analyzing the next sample. From these results, it is clear that even using high-sensitivity LC/MS analysis, low-carryover performance was maintained regardless of the polarity of the compound or its acidity / alkalinity. The excellent low-carryover performance of this system permits stable analysis without the influence of a preceding analysis on the subsequent analysis. 0.0 0.5 1.0 1.5 min 0 2.5 5.0 7.5 291.2(+) TIC(-) TIC(+) Inten. (×1,000,000) Trimethoprim Blank MS Chromatogram MS Spectrum Analysis Number ① ② log P : 0.91 pKa : 7.12 0.0 0.5 1.0 1.5 min 0 1 2 3 4 291.2(+) TIC(-) TIC(+) Inten. (×100,000) Sample Name 250.0 275.0 300.0 325.0 350.0 m/z 0 1 2 3 4 Inten. (×1,000,000) 291.2 Positive Operational efficiency is an important issue to researchers involved in the organic synthesis stage of drug discovery typically conducted at pharmaceutical companies and universities. Specifically, large numbers of drug candidates are synthesized daily in the drug discovery chemistry operations conducted throughout the pharmaceutical industry, and the synthesis and purity confirmation of these substances are primarily conducted by LC/MS. Typically, the LC/MS is treated as a shared instrument in an open-access environment, so rather than optimizing the analytical conditions for the various individual compounds, each type of compound is analyzed using a fixed set of analytical conditions. Further, the analyst conducting an analysis is often unaware of the type of compound that was analyzed by the previous analyst who used that instrument. Reliable analysis in such an environment fundamentally requires an LC system that ensures low carryover performance. The new Nexera-i integrated Ultra High Performance Liquid Chromatograph effectively suppresses carryover using specially treated liquid-contact surfaces, such as the needle seal and sampling needle, thereby providing excellent quantitative performance with high accuracy in a wide variety of analyses. Furthermore, the Nexera-i can be connected with the high-scan-speed LCMS-2020. The built-in UV / VIS detector or PDA detector along with connection to an LCMS permit very detailed qualitative and quantitative analysis. Here, we introduce an example of analysis of eight pharmaceutical substances in a workflow that is typically used for pharmaceutical synthesis confirmation analysis. Fig. 1.1 Chromatograms and Spectra of Eight Pharmaceutical Compounds and Blank Solutions
Transcript of by Nexera-i and LCMS-2020 No - Shimadzu · 2018. 11. 1. · LCMS-2020. The built-in UV/ VIS...
ApplicationNews
No.L465
High Performance Liquid Chromatography
Investigation of Synthetic Compounds in Drug Discovery by Nexera-i and LCMS-2020
LAAN-A-LC-E242
n Investigation of Synthetic Compounds in Open Access Environment
A workflow in which multiple analysts in an open access environment can quickly verify the synthesis of a variety of compounds using the same method is perceived as one of the solutions that can lead to greater efficiency in the research and development process. This Application News describes an example in which measurement was conducted for a range of compounds with properties spanning acidic to basic, and high to low polarity, all using the same analytical conditions (Table 1).Each of the various compounds was dissolved in dimethylsulfoxide (DMSO), and 0.1 µL was injected after diluting with methanol solution containing 5 % DMSO (Final concentration 20 mmol/L each). MS analysis was conducted using scan mode (m/z = 165 – 385) and positive / negative mode. The composition analysis results for the eight pharmaceutical compounds are shown below (Fig. 1.1 – 1.4). Also, assuming an actual workf low, we in jected a blank solut ion (methanol) prior to analyzing the next sample. From these results, it is clear that even using high-sensitivity LC/MS analysis, low-carryover performance was maintained regardless of the polarity of the compound or its acidity / alkalinity. The excellent low-carryover performance of this system permits stable analysis without the influence of a preceding analysis on the subsequent analysis.
0.0 0.5 1.0 1.5 min
0
2.5
5.0
7.5 291.2(+)TIC(-)TIC(+)Inten. (×1,000,000)
Trimethoprim
Blank
MS Chromatogram MS SpectrumAnalysis Number
①
②
log P : 0.91pKa : 7.12
0.0 0.5 1.0 1.5 min
0
1
2
3
4291.2(+)
TIC(-)TIC(+)Inten. (×100,000)
Sample Name
250.0 275.0 300.0 325.0 350.0 m/z0
1
2
3
4Inten. (×1,000,000)
291.2Positive
Operational efficiency is an important issue to researchers involved in the organic synthesis stage of drug discovery typically conducted at pharmaceutical companies and universities. Specifically, large numbers of drug candidates are synthesized dai ly in the drug discovery chemistry operations conducted throughout the pharmaceutical industry, and the synthesis and purity confirmation of these substances are primarily conducted by LC/MS. Typically, the LC/MS is treated as a shared instrument in an open-access environment, so rather than optimizing the analytical conditions for the various individual compounds, each type of compound is analyzed using a fixed set of analytical conditions. Further, the analyst conducting an analysis is often unaware of the type of compound that was analyzed by the previous analyst who used that instrument.Reliable analysis in such an environment fundamentally requires an LC system that ensures low carryover performance. The new Nexera-i integrated Ultra High Performance Liquid Chromatograph effectively suppresses carryover using specially treated liquid-contact surfaces, such as the needle seal and sampling needle, thereby providing excellent quantitative performance with high accuracy in a wide variety of analyses. Furthermore, the Nexera-i can be connected with the high-scan-speed LCMS-2020. The built-in UV / VIS detector or PDA detector along with connection to an LCMS permit very detailed qualitative and quantitative analysis.Here, we introduce an example of analysis of eight pharmaceutical substances in a workflow that is typically used for pharmaceutical synthesis confirmation analysis.
Fig. 1.1 Chromatograms and Spectra of Eight Pharmaceutical Compounds and Blank Solutions
ApplicationNews
No.L465
0.0 0.5 1.0 1.5 min
0
2.5
5
235.3(+)TIC(-)TIC(+)Inten. (×1,000,000)
0.0 0.5 1.0 1.5 min
0
1
2
3
4235.3(+)
TIC(-)TIC(+)
Inten. (×100,000)
0.0 0.5 1.0 1.5 min
0
5
10260.2(+)TIC(-)TIC(+)
Inten. (×1,000,000)
0.0 0.5 1.0 1.5 min
0
1
2
3
4260.2(+)
TIC(-)TIC(+)
Inten. (×100,000)
0.0 0.5 1.0 1.5 min
0
2.5
5.0
7.5
10
189.2(+)TIC(-)TIC(+)Inten. (×1,000,000)
0.0 0.5 1.0 1.5 min
0
1
2
3
4
5
189.2(+)
TIC(-)TIC(+)
Inten. (×100,000)
Blank
Blank
Blank
Lidocaine
Propranolol
Antipyrine
200.0 250.0 300.0 350.0 m/z0
1
2
3Inten. (×1,000,000)
189.2Positive
200.0 250.0 300.0 350.0 m/z0
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3
4
5
6
Inten. (×1,000,000)
260.2Positive
250.0 300.0 350.0 m/z0
1
2
3
4Inten. (×1,000,000)
235.3 Positive
log P : 2.44pKa : 8.01
log P : 3.48pKa : 9.42
log P : 0.38pKa : 1.4
⑧
③
④
⑤
⑥
⑦
MS Chromatogram MS SpectrumSample Name
Fig. 1.2 Chromatograms and Spectra of Eight Pharmaceutical Compounds and Blank Solutions
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No.L465
243.2(+)
Blank
Blank
Blank
Desipramine
Chlorpromazine
Bifonazole
0
1
2
3
log P : 4.9pKa : 10.4
log P : 5.41pKa : 9.3
log P : 4.77*pKa : 6.69
*Experimental Properties
MS Chromatogram MS SpectrumSample Name
⑨
⑩
⑪
⑫
⑬
⑭
1.5 min
Inten. (×10,000,000) Inten. (×1,000,000)
267.3(+)TIC(-)TIC(+)
267.3Positive
0
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1.0
1.5
200.0 250.0 300.0 350.00
1
2
3
4
5
Inten. (×100,000)
0
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4267.3(+)
TIC(-)TIC(+)
0
0.5
1.0
1.5 319.1(+)TIC(-)TIC(+)
Inten. (×10,000,000)
200.0 250.0 300.0 350.0
319.1Positive
0
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2
3
4319.1(+)
TIC(-)TIC(+)
Inten. (×100,000)
200.0 250.0 300.0 350.00
1
2
3
4
Inten. (×10,000,000) Inten. (×1,000,000)
0
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2311.3(+)
TIC(-)TIC(+)
311.3243.2
Positive
0.0 0.5 1.0
1.5 min0.0 0.5 1.0
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311.3(+)243.2(+)TIC(-)
TIC(+)
Inten. (×100,000)
Inten. (×1,000,000)
m/z
m/z
m/z
Fig. 1.3 Chromatograms and Spectra of Eight Pharmaceutical Compounds and Blank Solutions
ApplicationNews
No.
For Research Use Only. Not for use in diagnostic procedures.The content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of Shimadzu. The information contained herein is provided to you "as is" without warranty of any kind including without limitation warranties as to its accuracy or completeness. Shimadzu does not assume any responsibility or liability for any damage, whether direct or indirect, relating to the use of this publication. This publication is based upon the information available to Shimadzu on or before the date of publication, and subject to change without notice.
© Shimadzu Corporation, 2014www.shimadzu.com/an/
L465
First Edition: Aug. 2014
0.0 0.5 1.0 1.5 min
0
2.5
5.0237.2(+)TIC(-)TIC(+)
Inten. (×1,000,000)
0.0 0.5 1.0 1.5 min
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2.5
5.0237.2(+)TIC(-)TIC(+)
Inten.(×100,000)
Blank⑯
⑮
Carbamazepine
200.0 250.0 300.00
1
2Inten. (×1,000,000)
237.2Positive
350.0 m/z
log P : 2.45*pKa : 15.96
*Experimental Properties
MS Chromatogram MS SpectrumSample Name
[LC]Column : Shim-pack XR-ODSⅡ (50 mm L. × 2.0 mm I.D., 2.2 µm)Mobile Phase : A) 0.1 % Formic Acid in Water B) 0.1 % Formic Acid in AcetonitrileTime Program : B. Conc. 5 % (0 min) → 95 % (1 - 1.5 min) → 5 % (1.51 - 3 min)Flowrate : 1.0 mL/min Column Temp. : 40 °CInjection Volume : 0.1 µLRinse Solution : 0.2 % Formic Acid in MethanolRinse Mode : Before and After
[MS]Detection(MS) : LCMS-2020Probe Voltage : +4.5 kV (ESI-Positive Mode) -3.5 kV (ESI-Negative Mode)Nebulizing Gas Flow : 1.5 mL/minDrying Gas Flow : 15.0 L/minDL Temp. : 250 °CBlock Heater Temp. : 400 °CDL, Q-array Voltages : Default ValuesEvent Time : 0.03 secScan Range : m/z: 165-385
Fig. 1.4 Chromatograms and Spectra of Eight Pharmaceutical Compounds and Blank Solutions
Table 1 Analytical Conditions
n An Autosampler that Supports Open Access EnvironmentThe direct access mechanism of the Nexera-i permits individual access to separate sample racks, making it possible to load samples in a rack other than the one in which sample injection operations are being conducted for the current analysis (Fig. 2). This, in effect, permits an analyst to load samples for analysis without interrupting an ongoing analysis of another analyst, making it suitable for use as a joint-use instrument. The result is that instrument uti l ization efficiency is enhanced, thereby permitting a more efficient research and development process.Furthermore, synthetic products developed in a laboratory setting are almost always produced in small quantities. This is a scenario in which the Nexera-i autosampler excels, as it supports very small-volume injections, thereby permitting analysis of limited-volume samples.
Fig. 2 Direct Access for Each Sample Rack
[References]Drug Bank (http://www.drugbank.ca) April 2014
