Drug Development and Verification of Elemental Impurities Content in Cyclosporin Injection USP 250 mg/5 ml by Inductively Coupled Plasma Mass Spectroscopy (ICP-MS)

Received: 01-Nov-2022, Manuscript No. JDAR-22-80793; Editor assigned: 03-Nov-2022, Pre QC No. JDAR-22-80793 (PQ); Reviewed: 17-Nov-2022, QC No. JDAR-22-80793; Revised: 22-Nov-2022, Manuscript No. JDAR-22-80793 (R); Published: 29-Nov-2022, DOI: 10.4303/JDAR/236207

Introduction

ICP-MS (inductively coupled plasma mass spectrometry) is an analytical technique for measuring elements in biological fluids at trace levels i.e., low and ultra-low quantities. Although some laboratories still employ earlier techniques like atomic absorption and atomic emission, there has been a gradual trend toward ICP-MS, notably in the last decade. The ICP-MS technique has the following advantages over other analysis and that are

• Limits of detection are extremely low

• An extensive linear range

• Possibilities for detecting element isotope composition

• Multi-element character and a high sample throughput

• Allows for more sensitive results

In ICP-MS, Atomic elements are ionized after passing through a plasma source. These ions are then separated based on their mass [1,2].

Elemental impurities in a therapeutic product have been linked to potential safety and toxicological hazards, hence proper analysis of elemental impurities is critical to reducing patient exposure. “Any elements that are not supposed to be present in the final formulation of a medicinal product are known as elemental impurities”. The goal of pharmaceutical elemental analysis is to find contaminants that could contaminate the finished product. Risk assessment is now a top responsibility for all pharmaceutical firms to guarantee that all pharmaceutical product components and production techniques comply with requirements. This, however, can be a difficult task for producers, especially when all possible sources of contaminants are taken into account. Excipients, water, the active pharmaceutical substances themselves, as well as container systems and production methods, are examples of such sources. When a possible concern is found, further information is needed, and elemental impurity testing becomes the next obstacle.

Cyclosporine, the active ingredient in Cyclosporine injectable, USP, is an 11-amino-acid cyclic polypeptide immunosuppressant. Beauveria nivea, a fungus species, produces it as a metabolite. This article emphasizes on method development and verification of elemental impurity in Cyclosporine Injection by Inductively Coupled Plasma-Mass Spectroscopy (ICP-MS) [3,4].

Materials and Methods

Table 1: Specification limits.

Table 2: Details of standard and sample to be used.

Table 3: Details of instruments, reagents and chemicals used instrument.

Table 4: Reagents and chemicals.

Table 5: Instrument parameters.

Table 6: Internal standard used for elements.

Table 7: Preparation of standard stock solution (1000 ppm).

Table 8: Preparation of standard solution A.

Table 9: Preparation of internal standard solution.

Table 10: Preparation of linearity levels.

Table 11: Concentration of element in linearity level.

Description of analytical method used

Determination of elemental impurities ICH Class 1(Cd,Pb,As,Hg), Class 2A (Co,V,Ni), Class 3(Li,Sb,Cu) content in Cyclosporine injection USP 250 mg/5 mL by ICP-MS [5-8].

Diluent

Transfer 5.0 mL of Concentrated Nitric acid (67%-69%) solution and 5.0 mL of Hydrochloric acid (34%-37%) into 1000 mL volumetric flask dilute to volume with Purified water.

Preparation of sample/standard blank solution

Transfer 0.250 mL Nitric acid and 0.250 ml HCl, in 50 mL graduated centrifuge tube. Add 0.500 mL of internal standard solution (1 ppm) and dilute to volume up to the mark with purified water.

Preparation of test solution

Transfer 0.500 mL of Sample, 0.250 mL Nitric acid and 0.250 mL HCl in 50 mL graduated centrifuge tube. Add 0.500 mL of internal standard solution (1 ppm) and dilute up to the mark with purified water.

Results and Discussion

Aspirate Diluent followed by Linearity level solutions. Further aspirate Diluent, Sample blank, Test solution followed by Linearity level-5 solution as bracketing standard from Tables 12 and 13.

Table 12: System Suitability.

Table 13: % Recovery of Test solution Spiked at 100% level

System suitability criteria

1. Correlation coefficient should not be less than 0.99.

2. % Recovery for bracketing standard should be within 70.0-150.0.

Results:

1. System meets the system suitability criteria as specified in method of analysis.

2. % Recovery for each analyte was found within 70.0– 150.0 for 100% level.

Acceptance criteria:

1. System should meet the system suitability criteria as specified in method of analysis.

2. % Recovery for each analyte should be within 70.0– 150.0 for 100% level.

Method verification results

Various verification paramerter like Specificity, Limit of Quantitation, Linearity and Range, Precision, Accuracy are performed to determine the suitability of Cyclosporin injection.

Note: More than one parameter was performed at once with relevant sequence having common system suitability. Diluent sample can be interspersed more than one to avoid carryover from previous sample.

Specificity

Purpose: To demonstrate the ability of the method to assess the analyte unequivocally in presence of components which may be expected to be present.

Prepared Diluent, Linearity Levels, Sample/Standard blank and Test solution as per the Analytical method.

Preparation of test solution spiked at 100% level: Transferred 0.500 mL of Sample, 0.250 mL Nitric acid and 0.250 ml HCl in 50 mL volumetric flask, added 0.500 mL of Standard solution A and 0.500 mL of Internal standard solution (1 ppm) and diluted to volume up to the mark with purified water.

Results:

1. System meets the system suitability criteria as specified in method of analysis.

2. % Recovery for each analyte was found within 70.0– 150.0 for 100% level from Tables 14 and 15.

Table 14: System Suitability.

Table 15: Specificity %Recovery of Test solution Spiked at 100% level.

Acceptance criteria:

1. System should meet the system suitability criteria as specified in method of analysis.

2. % Recovery for each analyte should be within 70.0– 150.0 for 100% level.

Limit of quantitation

Purpose: To determine the lowest amount of an analyte in a sample that can be determined with acceptable precision and accuracy under the stated experimental conditions.

Prepared Diluent, Linearity Levels and Sample/Standard blank as per Analytical method.

Preparation of LOQ solution: Considered Linearity Level 70.0–150.02 as LOQ solution for determination from Tables 16-18.

Table 16: System Suitability.

Table 17: LOQ %RSD and concentration.

Table 18: Intensity (cps) of LOQ and Satandard/Sample Blank

Results:

1. System meets the system suitability criteria as specified in method of analysis.

2. % RSD of ratio of CPS of analyte of six LOQ solutions were found not more than 20.0.

3. Measured values for Blank was lower than the established Limit of Quantitation (LOQ) for each element.

Acceptance criteria:

1. System should meet the system suitability criteria as specified in method of analysis.

2. % RSD of ratio of CPS of analyte of six LOQ solutions should not be more than 20.0.

3. Measure values for Blank should be lower than the established Limit of Quantitation (LOQ) for each element.

Conclusion: % RSD of ratio of CPS of analyte of six LOQ solutions for all elements are well within acceptance criteria indicating that the method is precise at LOQ level.

Linearity and range

Purpose: To determine the Linearity and Range of the method.

The linearity of an analytical procedure is its ability (within a given range) to obtain test results which are directly proportional to the concentration (amount) of analyte in the sample. The range of an analytical procedure is the interval between the upper and lower concentration (amounts) of analyte in the sample (including these concentrations) for which it has been demonstrated that the analytical procedure has a suitable level of precision, accuracy and linearity.

Prepared Diluent, Linearity Levels and Sample/Standard blank as per Analytical method.

Results:

1. System meets the system suitability criteria as specified in method of analysis.

2. Correlation coefficient for each element were found not less than 0.99.

3. % RSD of ratio of CPS of analyte and Internal Standard of Six LOQ Level and Linearity Level 6 solutions were found not more than 20.0 from Tables 19–21.

Table 19: System Suitability

Table 20: Corelation Coefficient (R2), Slope and Intercept.

Table 21: %RSD of ratio of CPS at LOQ Level and Linearity Level 6.

Acceptance criteria:

1. System should meet the system suitability criteria as specified in method of analysis.

2. Correlation coefficient for each element should not be less than 0.99.

3. % RSD of ratio of CPS of analyte and Internal Standard of Six LOQ Level and Linearity Level 6 solutions should not be more than 20.0

Precision

Purpose: To demonstrate the repeatability under the same operating conditions over a short period of time.

System Precision was demonstrated by injecting the six Injections for Linearity Level 4 (100% Level).

Prepared Diluent, Linearity Levels and Sample/Standard blank as per Analytical method.

Results:

1. System meets the system suitability criteria as specified in method of analysis.

2. % RSD of ratio of analyte of six Linearity Level-4 solutions were found not more than 20.0.

Acceptance criteria:

1. System should meet the system suitability criteria as specified in method of analysis.

2. % RSD of ratio of CPS of analyte and Internal Standard of six Linearity Level-4 solutions should not be more than 20.0 from Tables 22 and 23.

Table 22: System Suitability.

Table 23: %RSD of ratio of CPS at Linearity Level 4.

Accuracy

Purpose: To measure the closeness of the test results obtained by a method to the true value. Accuracy study was performed at following levels by preparing three spike Test solutions each at LOQ 50%, 100% and 150% level.

Prepared Diluent, Linearity Levels, Sample/Standard blank and Test solution as per Analytical method as shown in Tables 24-28.

Table 24: System Suitability.

Table 25: %Recovery at LOQ level.

Table 26: %Recovery at 50% level.

Table 27: %Recovery at 100% level.

Table 28: %Recovery at 150% level.

Preparation of accuracy solution at LOQ level

Transfered 0.500 mL of Sample, 0.250 mL Nitric acid and 0.250 mL HCl in 50 mL volumetric flask, added 0.150 mL of Standard solution B and 0.500 mL of Internal standard solution (1 ppm) and diluted to volume up to the mark with purified water.

Prepared three Accuracy solutions at LOQ level as per above mentioned procedure and labeled as Accuracy Level LOQ_Prep- 1, Accuracy Level LOQ_Prep-2, and Accuracy Level LOQ_ Prep-3.

Preparation of accuracy solution at 50% level

Transfered 0.500 mL of Sample, 0.250 mL Nitric acid and 0.250 ml HCl in 50 mL volumetric flask, added 0.250 mL of Standard solution B and 0.500 mL of Internal standard solution (1 ppm) and diluted to volume up to the mark with purified water.

Prepared three Accuracy solutions at 50% level as per above mentioned procedure and labeled as Accuracy Level 50%_Prep- 1, Accuracy Level 50%_Prep-2, and Accuracy Level 50%_Prep- 3.

Preparation of accuracy solution at 100% level

Transfered 0.500 mL of Sample, 0.250 mL Nitric acid and 0.250 mL HCl in 50 mL volumetric flask, added 0.500 mL of Standard solution B and 0.500 mL of Internal standard solution (1 ppm) and diluted to volume up to the mark with purified water.

Prepared three Accuracy solutions at 100% level as per above mentioned procedure and labeled as Accuracy Level 100%_Prep-1, Accuracy Level 100%_Prep-2, and Accuracy Level 100%_Prep-3.

Preparation of accuracy solution at 150% level

Transfered 0.500 mL of Sample, 0.250 mL Nitric acid and 0.250 mL HCl in 50 mL volumetric flask, added 0.750 mL of Standard solution B and 0.500 mL of Internal standard solution (1 ppm) and diluted to volume up to the mark with purified water.

Prepared three Accuracy solutions at 150% level as per above mentioned procedure and labeled as Accuracy Level 150%_Prep-1, Accuracy Level 150%_Prep-2, and Accuracy Level 150%_Prep-3 as shown in Tables 29 and 30.

Table 29: Abbreviation used in this project.

Table 30: Summary of results.

Results:

1. System meets the system suitability criteria as specified in method of analysis.

2. Each % Recovery for the analyte were found within 70.0–150.0 for all linearity levels.

Acceptance criteria:

1. System should meet the system suitability criteria as specified in method of analysis.

2. % Recovery for each element should be within 70.0– 150.0 for all accuracy levels.

Conclusion

Based upon the data and results obtained for the analytical method verification, the ICP-MS method used for the determination of elemental impurities in Cyclosporine injection USP 250 mg/5 ml specific and selective. This method is linear over the range of 10% to 200%. The method is precise. The ICP-MS method for the evaluation elemental impurities in Cyclosporine injection USP 250 mg/5 mL has been verified.

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Copyright: © 2022 Smruti Ranjan Mohanty, et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.