Impurities in pharmaceuticals must be appropriately controlled to ensure product safety. The ICH guidelines on impurities in pharmaceutical substances 1) and impurities in drug products 2) have established specific thresholds for impurities. If the content of impurity in pharmaceuticals is more than 0.1%, it is necessary to identify its structure, perform the safety evaluation and report to the regulatory authorities.

Therefore, the accurate determination of trace impurities is of great importance in both pharmaceutical drug development and drug manufacturing. The following procedure is commonly used to measure the concentration of impurities in

pharmaceutical products using area percentage normalization: prepare and analyze the

standard solution at a concentration such that the height of the main peak

will be within the linear range of the detector, then analyze multiple dilutions of the sample. Since the new photodiode array detector SPD-M40 of N-Series UHPLC has completely eliminated the effects of stray light during detection, it not only provides a wide dynamic range (linearity up to 2.5 AU as a specification value, typical value is more than 2.5 AU) but also achieves low noise and high sensitivity.

Moreover, the SPD-M40 has little fluctuation in its baseline

thanks to Advanced TC-Optics (triple temperature control of the

cell, light source, and optical system), making it ideal for analyzing

trace impurities in pharmaceuticals.

Phar

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64 Spinco Biotech

Analysis of Impurities in Pharmaceutical Products - using N-Series UHPLC /PDA with Shim-pack Velox C18 Column

Application News

No. L538

High Performance Liquid Chromatography

Impurity Analysis in Pharmaceutical Products with the Advanced Photodiode Array Detector SPD-M40

LAAN-A-LC-E311

Impurities in pharmaceuticals must be appropriately controlled to ensure product safety. The ICH guidelines on impurities in pharmaceutical substances 1) and impurities in drug products 2) have established specific thresholds for impurities. If the content of impurity in pharmaceuticals is more than 0.1 %, it is necessary to identify its structure, perform the safety evaluation and report to the regulatory authorities. Therefore, the accurate determination of trace impurities is of great importance in both pharmaceutical drug development and drug manufacturing. The following procedure is commonly used to measure the concentration of impurities in pharmaceutical products using area percentage normalization: prepare and analyze the standard solution at a concentration such that the height of the main peak will be within the linear range of the detector, then analyze multiple dilutions of the sample. Since the new photodiode array detector SPD-M40 has completely eliminated the effects of stray light during detection, it not only provides a wide dynamic range (linearity up to 2.5 AU as a specification value, typical value is more than 2.5 AU) but also achieves low noise and high sensitivity. Moreover, the SPD-M40 has little fluctuation in its baseline thanks to Advanced TC-Optics (triple temperature control of the cell, light source, and optical system), making it ideal for analyzing trace impurities in pharmaceuticals.

Y. Zhou, H. Terada

Table 1 Analytical Conditions Column : Shim-pack Velox™ C18

(100 mm L. × 3.0 mm I.D., 2.7 μm) Mode : Low pressure gradient Mobile Phase : A) 10 mM Sodium phosphate buffer (pH=2.6) B) Acetonitrile Flow Rate : 1 mL/min Column Temp. : 40 °C Injection Volume : 2 μL Detection : SPD-M40 at 256 nm Flow Cell : Standard cell

Table 2 Gradient Time Program

Time (min) A. Conc B. Conc 0 70 30 4 40 60 6 10 90 8 10 90

8.01 70 30 10 STOP

Linearity This section presents examples of impurity analyses with SPD-M40 using ketoprofen, a nonsteroidal anti-inflammatory drug. Analytical conditions and gradient time programs for mobile phases are shown in Table 1 and Table 2, respectively. To confirm the linearity of ketoprofen with SPD-M40, 0.5-800 mg/L of standard solutions were prepared and analyzed. Fig. 1 shows the chromatograms of the ketoprofen standard solutions, and Fig. 2 shows the calibration curve of ketoprofen. Excellent linearity (R2 ≥ 0.999) was obtained over a wide range of concentrations, from 0.5 to 800 mg/L.

Fig. 1 Chromatograms of the Ketoprofen Standard Samples

Fig. 2 Calibration Curve for Ketoprofen

Ketoprofen0.5 ~ 800 mg/L

2.25 2.50 2.75 3.00 3.25 min

0.0

0.5

1.0

1.5

2.0

2.5

3.0

AU

Zoomed-in

2.4 2.5 2.6 min

0

2

4

6

8

mAU

0 250 500 750 Concentration (mg/L)0.00

0.50

1.00

1.50

2.00

2.50

3.00Peak height (AU)

R2: 0.9998

Application News

No. L538

High Performance Liquid Chromatography

Impurity Analysis in Pharmaceutical Products with the Advanced Photodiode Array Detector SPD-M40

LAAN-A-LC-E311

Impurities in pharmaceuticals must be appropriately controlled to ensure product safety. The ICH guidelines on impurities in pharmaceutical substances 1) and impurities in drug products 2) have established specific thresholds for impurities. If the content of impurity in pharmaceuticals is more than 0.1 %, it is necessary to identify its structure, perform the safety evaluation and report to the regulatory authorities. Therefore, the accurate determination of trace impurities is of great importance in both pharmaceutical drug development and drug manufacturing. The following procedure is commonly used to measure the concentration of impurities in pharmaceutical products using area percentage normalization: prepare and analyze the standard solution at a concentration such that the height of the main peak will be within the linear range of the detector, then analyze multiple dilutions of the sample. Since the new photodiode array detector SPD-M40 has completely eliminated the effects of stray light during detection, it not only provides a wide dynamic range (linearity up to 2.5 AU as a specification value, typical value is more than 2.5 AU) but also achieves low noise and high sensitivity. Moreover, the SPD-M40 has little fluctuation in its baseline thanks to Advanced TC-Optics (triple temperature control of the cell, light source, and optical system), making it ideal for analyzing trace impurities in pharmaceuticals.

Y. Zhou, H. Terada

Table 1 Analytical Conditions Column : Shim-pack Velox™ C18

(100 mm L. × 3.0 mm I.D., 2.7 μm) Mode : Low pressure gradient Mobile Phase : A) 10 mM Sodium phosphate buffer (pH=2.6) B) Acetonitrile Flow Rate : 1 mL/min Column Temp. : 40 °C Injection Volume : 2 μL Detection : SPD-M40 at 256 nm Flow Cell : Standard cell

Table 2 Gradient Time Program

Time (min) A. Conc B. Conc 0 70 30 4 40 60 6 10 90 8 10 90

8.01 70 30 10 STOP

Linearity This section presents examples of impurity analyses with SPD-M40 using ketoprofen, a nonsteroidal anti-inflammatory drug. Analytical conditions and gradient time programs for mobile phases are shown in Table 1 and Table 2, respectively. To confirm the linearity of ketoprofen with SPD-M40, 0.5-800 mg/L of standard solutions were prepared and analyzed. Fig. 1 shows the chromatograms of the ketoprofen standard solutions, and Fig. 2 shows the calibration curve of ketoprofen. Excellent linearity (R2 ≥ 0.999) was obtained over a wide range of concentrations, from 0.5 to 800 mg/L.

Fig. 1 Chromatograms of the Ketoprofen Standard Samples

Fig. 2 Calibration Curve for Ketoprofen

Ketoprofen0.5 ~ 800 mg/L

2.25 2.50 2.75 3.00 3.25 min

0.0

0.5

1.0

1.5

2.0

2.5

3.0

AU

Zoomed-in

2.4 2.5 2.6 min

0

2

4

6

8

mAU

0 250 500 750 Concentration (mg/L)0.00

0.50

1.00

1.50

2.00

2.50

3.00Peak height (AU)

R2: 0.9998

Application News

No. L538

High Performance Liquid Chromatography

Impurity Analysis in Pharmaceutical Products with the Advanced Photodiode Array Detector SPD-M40

LAAN-A-LC-E311

Impurities in pharmaceuticals must be appropriately controlled to ensure product safety. The ICH guidelines on impurities in pharmaceutical substances 1) and impurities in drug products 2) have established specific thresholds for impurities. If the content of impurity in pharmaceuticals is more than 0.1 %, it is necessary to identify its structure, perform the safety evaluation and report to the regulatory authorities. Therefore, the accurate determination of trace impurities is of great importance in both pharmaceutical drug development and drug manufacturing. The following procedure is commonly used to measure the concentration of impurities in pharmaceutical products using area percentage normalization: prepare and analyze the standard solution at a concentration such that the height of the main peak will be within the linear range of the detector, then analyze multiple dilutions of the sample. Since the new photodiode array detector SPD-M40 has completely eliminated the effects of stray light during detection, it not only provides a wide dynamic range (linearity up to 2.5 AU as a specification value, typical value is more than 2.5 AU) but also achieves low noise and high sensitivity. Moreover, the SPD-M40 has little fluctuation in its baseline thanks to Advanced TC-Optics (triple temperature control of the cell, light source, and optical system), making it ideal for analyzing trace impurities in pharmaceuticals.

Y. Zhou, H. Terada

Table 1 Analytical Conditions Column : Shim-pack Velox™ C18

(100 mm L. × 3.0 mm I.D., 2.7 μm) Mode : Low pressure gradient Mobile Phase : A) 10 mM Sodium phosphate buffer (pH=2.6) B) Acetonitrile Flow Rate : 1 mL/min Column Temp. : 40 °C Injection Volume : 2 μL Detection : SPD-M40 at 256 nm Flow Cell : Standard cell

Table 2 Gradient Time Program

Time (min) A. Conc B. Conc 0 70 30 4 40 60 6 10 90 8 10 90

8.01 70 30 10 STOP

Linearity This section presents examples of impurity analyses with SPD-M40 using ketoprofen, a nonsteroidal anti-inflammatory drug. Analytical conditions and gradient time programs for mobile phases are shown in Table 1 and Table 2, respectively. To confirm the linearity of ketoprofen with SPD-M40, 0.5-800 mg/L of standard solutions were prepared and analyzed. Fig. 1 shows the chromatograms of the ketoprofen standard solutions, and Fig. 2 shows the calibration curve of ketoprofen. Excellent linearity (R2 ≥ 0.999) was obtained over a wide range of concentrations, from 0.5 to 800 mg/L.

Fig. 1 Chromatograms of the Ketoprofen Standard Samples

Fig. 2 Calibration Curve for Ketoprofen

Ketoprofen0.5 ~ 800 mg/L

2.25 2.50 2.75 3.00 3.25 min

0.0

0.5

1.0

1.5

2.0

2.5

3.0

AU

Zoomed-in

2.4 2.5 2.6 min

0

2

4

6

8

mAU

0 250 500 750 Concentration (mg/L)0.00

0.50

1.00

1.50

2.00

2.50

3.00Peak height (AU)

R2: 0.9998

Fig. 1: Chromatograms of the Ketoprofen Standard Samples

Fig. 2: Calibration Curve for Ketoprofen

Application News

No. L538

High Performance Liquid Chromatography

Impurity Analysis in Pharmaceutical Products with the Advanced Photodiode Array Detector SPD-M40

LAAN-A-LC-E311

Impurities in pharmaceuticals must be appropriately controlled to ensure product safety. The ICH guidelines on impurities in pharmaceutical substances 1) and impurities in drug products 2) have established specific thresholds for impurities. If the content of impurity in pharmaceuticals is more than 0.1 %, it is necessary to identify its structure, perform the safety evaluation and report to the regulatory authorities. Therefore, the accurate determination of trace impurities is of great importance in both pharmaceutical drug development and drug manufacturing. The following procedure is commonly used to measure the concentration of impurities in pharmaceutical products using area percentage normalization: prepare and analyze the standard solution at a concentration such that the height of the main peak will be within the linear range of the detector, then analyze multiple dilutions of the sample. Since the new photodiode array detector SPD-M40 has completely eliminated the effects of stray light during detection, it not only provides a wide dynamic range (linearity up to 2.5 AU as a specification value, typical value is more than 2.5 AU) but also achieves low noise and high sensitivity. Moreover, the SPD-M40 has little fluctuation in its baseline thanks to Advanced TC-Optics (triple temperature control of the cell, light source, and optical system), making it ideal for analyzing trace impurities in pharmaceuticals.

Y. Zhou, H. Terada

Table 1 Analytical Conditions Column : Shim-pack Velox™ C18

(100 mm L. × 3.0 mm I.D., 2.7 μm) Mode : Low pressure gradient Mobile Phase : A) 10 mM Sodium phosphate buffer (pH=2.6) B) Acetonitrile Flow Rate : 1 mL/min Column Temp. : 40 °C Injection Volume : 2 μL Detection : SPD-M40 at 256 nm Flow Cell : Standard cell

Table 2 Gradient Time Program

Time (min) A. Conc B. Conc 0 70 30 4 40 60 6 10 90 8 10 90

8.01 70 30 10 STOP

Linearity This section presents examples of impurity analyses with SPD-M40 using ketoprofen, a nonsteroidal anti-inflammatory drug. Analytical conditions and gradient time programs for mobile phases are shown in Table 1 and Table 2, respectively. To confirm the linearity of ketoprofen with SPD-M40, 0.5-800 mg/L of standard solutions were prepared and analyzed. Fig. 1 shows the chromatograms of the ketoprofen standard solutions, and Fig. 2 shows the calibration curve of ketoprofen. Excellent linearity (R2 ≥ 0.999) was obtained over a wide range of concentrations, from 0.5 to 800 mg/L.

Fig. 1 Chromatograms of the Ketoprofen Standard Samples

Fig. 2 Calibration Curve for Ketoprofen

Ketoprofen0.5 ~ 800 mg/L

2.25 2.50 2.75 3.00 3.25 min

0.0

0.5

1.0

1.5

2.0

2.5

3.0

AU

Zoomed-in

2.4 2.5 2.6 min

0

2

4

6

8

mAU

0 250 500 750 Concentration (mg/L)0.00

0.50

1.00

1.50

2.00

2.50

3.00Peak height (AU)

R2: 0.9998

Table 1: Analytical Conditions

Table 2: Gradient Time Program

Application Brief

July 2019

Phar

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65Spinco Biotech

LinearityThis section presents examples of impurity analyses with SPD-M40 using ketoprofen, a nonsteroidal antiinflammatory drug.

Analytical conditions and gradient time programs for mobile phases are shown in Table 1 and Table 2, respectively.

To confirm the linearity of ketoprofen with SPD-M40, 0.5-800 mg/L of standard solutions were prepared and analyzed. Figure 1 shows the chromatograms of the ketoprofen standard solutions, and Figure 2 shows the calibration curve of ketoprofen.

Excellent linearity (R2 ≥ 0.999) was obtained over a wide range of concentrations, from 0.5 to 800 mg/L.

Impurity analysisFigure 3 shows the chromatograms of six repeated analyses of 700 mg/L ketoprofen standard solution with peak heights of approximately 2.5 AU. Table 3 shows the area percentage and area coefficient of variation (%RSD) of ketoprofen and each impurity component. The peak area RSD% of Impurity 1 (the only impurity with content >0.1%) was less than 1%, showing good reproducibility.

Peak purity and spectral confirmationOne of the main advantages of PDA detection for impurity analysis is the possibility of performing a peak purity assay and impurity spectral confirmation. Figure 4 shows the purity curve of the ketoprofen peak. No impurities were detected within the peak elution interval. Figure 5 shows the UV spectrum of Impurity 3 with an area percentage concentration of 0.046 %. This result shows that informative spectra can be obtained even at very low concentrations.

Results for pharmaceutical analyses obtained using SPD-M40 are highly reliable due to its high performance over a wide range of concentrations.

References1) Revision of Guidelines for Impurities in New Drug Substances

(Evaluation and Licensing Division, PMSB, dated December 16, 2002, Notification No. 1216001).

2) Revision of Guidelines for Impurities in New Drug Substances (Evaluation and Licensing Division, PMSB dated June 24, 2003, Notification No. 0624001).

Application News

No. L538

First Edition: Apr. 2019

For Research Use Only. Not for use in diagnostic procedure.

This publication may contain references to products that are not available in your country. Please contact us to check the availability of these products in your country. The content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of Shimadzu. Shimadzu disclaims any proprietary interest in trademarks and trade names used in this publication other than its own. See http://www.shimadzu.com/about/trademarks/index.html for details. 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, 2019

www.shimadzu.com/an/

Fig. 3 Overlapped Chromatograms of Ketoprofen 700 mg/L Standard Solution (n = 6)

Impurity Analysis Fig. 3 shows the chromatograms of six repeated analyses of 700 mg/L ketoprofen standard solution with peak heights of approximately 2.5 AU. Table 3 shows the area percentage and area coefficient of variation (%RSD) of ketoprofen and each impurity component. The peak area RSD% of Impurity 1 (the only impurity with content >0.1 %) was less than 1 %, showing good reproducibility.

Table 3 Analytical Results for Each Component No. Compounds Retention Time (min) Area (%) Area (%RSD)

1 Ketoprofen 2.583 99.704 0.001 2 Impurity 1 1.998 0.126 0.598 3 Impurity 2 3.072 0.075 0.543 4 Impurity 3 3.186 0.046 1.115 5 Impurity 4 3.834 0.025 1.644 6 Impurity 5 4.446 0.011 4.556 7 Impurity 6 5.118 0.012 3.355

Peak Purity and Spectral Confirmation One of the main advantages of PDA detection for impurity analysis is the possibility of performing a peak purity assay and impurity spectral confirmation. Fig. 4 shows the purity curve of the ketoprofen peak. No impurities were detected within the peak elution interval. Fig. 5 shows the UV spectrum of Impurity 3 with an area percentage concentration of 0.046 %. This result shows that informative spectra can be obtained even at very low concentrations. Results for pharmaceutical analyses obtained using SPD-M40 are highly reliable due to its high performance over a wide range of concentrations.

Fig. 4 Purity Curve of the Ketoprofen Peak

Fig. 5 UV Spectrum of Impurity 3

References 1) Revision of Guidelines for Impurities in New Drug Substances

(Evaluation and Licensing Division, PMSB, dated December 16, 2002, Notification No. 1216001).

2) Revision of Guidelines for Impurities in New Drug Substances (Evaluation and Licensing Division, PMSB dated June 24, 2003, Notification No. 0624001).

Shim-pack Velox is a trademark of Shimadzu Corporation in Japan and/or other countries.

IMP-1

IMP-2

IMP-3IMP-4 IMP-5 IMP-6

Ketoprofen

0.0 1.0 2.0 3.0 4.0 5.0 Min

0

2

4

6

8

10

12

mAU

2.55 2.60 2.65 2.70 2.75 min

0.00

0.25

0.50

0.75

0

500

1000

1500

2000

2500

mAUPeak

Zero auxiliary linePurity curve

200 225 250 275 300 325 Nm

0.0

1.0

2.0

3.0mAU

234

260

Application News

No. L538

First Edition: Apr. 2019

For Research Use Only. Not for use in diagnostic procedure.

This publication may contain references to products that are not available in your country. Please contact us to check the availability of these products in your country. The content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of Shimadzu. Shimadzu disclaims any proprietary interest in trademarks and trade names used in this publication other than its own. See http://www.shimadzu.com/about/trademarks/index.html for details. 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, 2019

www.shimadzu.com/an/

Fig. 3 Overlapped Chromatograms of Ketoprofen 700 mg/L Standard Solution (n = 6)

Impurity Analysis Fig. 3 shows the chromatograms of six repeated analyses of 700 mg/L ketoprofen standard solution with peak heights of approximately 2.5 AU. Table 3 shows the area percentage and area coefficient of variation (%RSD) of ketoprofen and each impurity component. The peak area RSD% of Impurity 1 (the only impurity with content >0.1 %) was less than 1 %, showing good reproducibility.

Table 3 Analytical Results for Each Component No. Compounds Retention Time (min) Area (%) Area (%RSD)

1 Ketoprofen 2.583 99.704 0.001 2 Impurity 1 1.998 0.126 0.598 3 Impurity 2 3.072 0.075 0.543 4 Impurity 3 3.186 0.046 1.115 5 Impurity 4 3.834 0.025 1.644 6 Impurity 5 4.446 0.011 4.556 7 Impurity 6 5.118 0.012 3.355

Peak Purity and Spectral Confirmation One of the main advantages of PDA detection for impurity analysis is the possibility of performing a peak purity assay and impurity spectral confirmation. Fig. 4 shows the purity curve of the ketoprofen peak. No impurities were detected within the peak elution interval. Fig. 5 shows the UV spectrum of Impurity 3 with an area percentage concentration of 0.046 %. This result shows that informative spectra can be obtained even at very low concentrations. Results for pharmaceutical analyses obtained using SPD-M40 are highly reliable due to its high performance over a wide range of concentrations.

Fig. 4 Purity Curve of the Ketoprofen Peak

Fig. 5 UV Spectrum of Impurity 3

References 1) Revision of Guidelines for Impurities in New Drug Substances

(Evaluation and Licensing Division, PMSB, dated December 16, 2002, Notification No. 1216001).

2) Revision of Guidelines for Impurities in New Drug Substances (Evaluation and Licensing Division, PMSB dated June 24, 2003, Notification No. 0624001).

Shim-pack Velox is a trademark of Shimadzu Corporation in Japan and/or other countries.

IMP-1

IMP-2

IMP-3IMP-4 IMP-5 IMP-6

Ketoprofen

0.0 1.0 2.0 3.0 4.0 5.0 Min

0

2

4

6

8

10

12

mAU

2.55 2.60 2.65 2.70 2.75 min

0.00

0.25

0.50

0.75

0

500

1000

1500

2000

2500

mAUPeak

Zero auxiliary linePurity curve

200 225 250 275 300 325 Nm

0.0

1.0

2.0

3.0mAU

234

260

Application News

No. L538

First Edition: Apr. 2019

For Research Use Only. Not for use in diagnostic procedure.

This publication may contain references to products that are not available in your country. Please contact us to check the availability of these products in your country. The content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of Shimadzu. Shimadzu disclaims any proprietary interest in trademarks and trade names used in this publication other than its own. See http://www.shimadzu.com/about/trademarks/index.html for details. 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, 2019

www.shimadzu.com/an/

Fig. 3 Overlapped Chromatograms of Ketoprofen 700 mg/L Standard Solution (n = 6)

Impurity Analysis Fig. 3 shows the chromatograms of six repeated analyses of 700 mg/L ketoprofen standard solution with peak heights of approximately 2.5 AU. Table 3 shows the area percentage and area coefficient of variation (%RSD) of ketoprofen and each impurity component. The peak area RSD% of Impurity 1 (the only impurity with content >0.1 %) was less than 1 %, showing good reproducibility.

Table 3 Analytical Results for Each Component No. Compounds Retention Time (min) Area (%) Area (%RSD)

1 Ketoprofen 2.583 99.704 0.001 2 Impurity 1 1.998 0.126 0.598 3 Impurity 2 3.072 0.075 0.543 4 Impurity 3 3.186 0.046 1.115 5 Impurity 4 3.834 0.025 1.644 6 Impurity 5 4.446 0.011 4.556 7 Impurity 6 5.118 0.012 3.355

Peak Purity and Spectral Confirmation One of the main advantages of PDA detection for impurity analysis is the possibility of performing a peak purity assay and impurity spectral confirmation. Fig. 4 shows the purity curve of the ketoprofen peak. No impurities were detected within the peak elution interval. Fig. 5 shows the UV spectrum of Impurity 3 with an area percentage concentration of 0.046 %. This result shows that informative spectra can be obtained even at very low concentrations. Results for pharmaceutical analyses obtained using SPD-M40 are highly reliable due to its high performance over a wide range of concentrations.

Fig. 4 Purity Curve of the Ketoprofen Peak

Fig. 5 UV Spectrum of Impurity 3

References 1) Revision of Guidelines for Impurities in New Drug Substances

(Evaluation and Licensing Division, PMSB, dated December 16, 2002, Notification No. 1216001).

2) Revision of Guidelines for Impurities in New Drug Substances (Evaluation and Licensing Division, PMSB dated June 24, 2003, Notification No. 0624001).

Shim-pack Velox is a trademark of Shimadzu Corporation in Japan and/or other countries.

IMP-1

IMP-2

IMP-3IMP-4 IMP-5 IMP-6

Ketoprofen

0.0 1.0 2.0 3.0 4.0 5.0 Min

0

2

4

6

8

10

12

mAU

2.55 2.60 2.65 2.70 2.75 min

0.00

0.25

0.50

0.75

0

500

1000

1500

2000

2500

mAUPeak

Zero auxiliary linePurity curve

200 225 250 275 300 325 Nm

0.0

1.0

2.0

3.0mAU

234

260

Application News

No. L538

First Edition: Apr. 2019

For Research Use Only. Not for use in diagnostic procedure.

This publication may contain references to products that are not available in your country. Please contact us to check the availability of these products in your country. The content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of Shimadzu. Shimadzu disclaims any proprietary interest in trademarks and trade names used in this publication other than its own. See http://www.shimadzu.com/about/trademarks/index.html for details. 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, 2019

www.shimadzu.com/an/

Fig. 3 Overlapped Chromatograms of Ketoprofen 700 mg/L Standard Solution (n = 6)

Impurity Analysis Fig. 3 shows the chromatograms of six repeated analyses of 700 mg/L ketoprofen standard solution with peak heights of approximately 2.5 AU. Table 3 shows the area percentage and area coefficient of variation (%RSD) of ketoprofen and each impurity component. The peak area RSD% of Impurity 1 (the only impurity with content >0.1 %) was less than 1 %, showing good reproducibility.

Table 3 Analytical Results for Each Component No. Compounds Retention Time (min) Area (%) Area (%RSD)

1 Ketoprofen 2.583 99.704 0.001 2 Impurity 1 1.998 0.126 0.598 3 Impurity 2 3.072 0.075 0.543 4 Impurity 3 3.186 0.046 1.115 5 Impurity 4 3.834 0.025 1.644 6 Impurity 5 4.446 0.011 4.556 7 Impurity 6 5.118 0.012 3.355

Peak Purity and Spectral Confirmation One of the main advantages of PDA detection for impurity analysis is the possibility of performing a peak purity assay and impurity spectral confirmation. Fig. 4 shows the purity curve of the ketoprofen peak. No impurities were detected within the peak elution interval. Fig. 5 shows the UV spectrum of Impurity 3 with an area percentage concentration of 0.046 %. This result shows that informative spectra can be obtained even at very low concentrations. Results for pharmaceutical analyses obtained using SPD-M40 are highly reliable due to its high performance over a wide range of concentrations.

Fig. 4 Purity Curve of the Ketoprofen Peak

Fig. 5 UV Spectrum of Impurity 3

References 1) Revision of Guidelines for Impurities in New Drug Substances

(Evaluation and Licensing Division, PMSB, dated December 16, 2002, Notification No. 1216001).

2) Revision of Guidelines for Impurities in New Drug Substances (Evaluation and Licensing Division, PMSB dated June 24, 2003, Notification No. 0624001).

Shim-pack Velox is a trademark of Shimadzu Corporation in Japan and/or other countries.

IMP-1

IMP-2

IMP-3IMP-4 IMP-5 IMP-6

Ketoprofen

0.0 1.0 2.0 3.0 4.0 5.0 Min

0

2

4

6

8

10

12

mAU

2.55 2.60 2.65 2.70 2.75 min

0.00

0.25

0.50

0.75

0

500

1000

1500

2000

2500

mAUPeak

Zero auxiliary linePurity curve

200 225 250 275 300 325 Nm

0.0

1.0

2.0

3.0mAU

234

260

Fig. 3: Overlapped Chromatograms of Ketoprofen 700 mg/L Standard Solution (n = 6)

Table 3: Analytical Results for Each Component

Fig. 4: Purity Curve of the Ketoprofen Peak

Fig. 5: UV Spectrum of Impurity 3

Application Brief

July 2019