Stability indicating RP-UPLC method for the simultaneous estimation of Niraparib and Abiraterone in pharmaceutical dosage forms
Shanthi Priya DK1,2*, Mukthinuthalapati
Mathrusri Annapurna1
1Department of
Pharmaceutical Analysis, GITAM School of Pharmacy, Visakhapatnam, India.
2School of Pharmacy, Anurag University, Ghatkesar, Hyderabad.
*Corresponding Author E-mail: sdarga@gitam.in
ABSTRACT:
Niraparib and Abiraterone are anti-cancer agents. A new stability indicating UPLC method has been developed and validated for the simultaneous assay of Niraparib and Abiraterone using Acquity UPLC HSS T3 Column with UV detection at 215 nm within a run time of 3 minutes where Niraparib was eluted at 1.169 min and that of Abiraterone at 0.684 min. Linearity was observed over the concentration range of 2.5-15 µg/mL for Niraparib and 12.5-75 µg/mL for Abiraterone respectively and the regression equations were found to be y=68577x+10960 and y=66848x+7064.7 for Niraparib and Abiraterone respectively. The LOD was found to be 0.32 μg/ml and 0.46 μg/ml whereas the LOQ was found to be 0.98 μg/ml and 1.38 μg/ml for Niraparib and Abiraterone respectively.
KEYWORDS: Niraparib, Abiraterone, UPLC, Validation, Forced degradation studies, ICH guidelines.
INTRODUCTION:
Niraparib (C26H30N4O5S), a PARP inhibitor is used in the treatment of ovarian cancer1. Niraparib is chemically Niraparib tosylate monohydrate which is 2-{4-[(3S)-piperidin-3-yl] phenyl}-2Hindazole 7-carboxamide 4-methyl benzenesulfonate hydrate (1:1:1) with molecular weight of 510.61 amu2 (Figure 1A). Abiraterone2 (C26H33NO2) is a derivative of steroidal progesterone and is an innovative drug that offers clinical benefit to patients with hormone refractory prostate cancer. Abiraterone is chemically (3β)-17-(pyridin-3-yl) androsta-5, 16-dien-3-ol with molecular weight of 391.55 (Figure 1B). Niraparib was estimated by using HPLC3, LC-MS/MS4-5 and Abiraterone by UPLC6, HPLC7-11 in pharmaceutical dosage forms as well as biological fluids. A new stability indicating RP-UPLC technique has been developed and validated for the simultaneous assay of Niraparib and Abiraterone
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Figure 1A: Structure of Niraparib |
Figure 1B: Structure of Abiraterone |
MATERIALS AND METHODS:
Acquity UPLC HSS T3 Column with Empower 2 software, UV detector, auto-injector, Shimadzu UV-Visible spectrophotometer, Ohus Electronic balance, Eutech pH Meter and Phoenix 4.5 L digital ultrasonic cleaner were used for the present study. Niraparib and Abiraterone API samples obtained from Pharma Life Research facility in Hyderabad, India and other chemicals were of AR-grade (Rankem Chemicals, India). Accurately weighed quantities of 10 mg Niraparib and 50 mg of Abiraterone were transferred into a clean, dry 100 mL volumetric flasks and diluted as per the requirement with the mobile phase.
Method validation12
The process of method validation involves conducting a series of tests that are based on the analytical method. These tests are used to determine and document the performance characteristics of the method, as well as to determine if the method is suitable for a specific analytical purpose. The performance characteristics of LC methods include specificity, selectivity, precision, linearity, robustness, recovery, range, limit of detection, limit of quantification, and ruggedness. The focus of validation should be on the precision of the analytical system, rather than just the method itself. This includes having a well-defined method protocol, concentration ranges for the analyte, and a specified type of test material. The method validation protocol will be prepared by carefully considering the analytical system as a whole and the analytical procedure.
Assay of Niraparib and Abiraterone
The combination of Niraparib and Abiraterone is available with the brand name Akeega with label claim 100 mg and 500 mg respectively.10 tablets of this marketed formulation were weighed, crushed in to a fine powder and powder equivalent to 100 mg Niraparib and 500 mg Abiraterone was transferred in to a volumetric flask containing acetonitrile and sonicated for 30 mins. The mixture was filtered and diluted with the mobile phase as per the requirement. A solution consisting of 10 μg/mL of Niraparib and 50 μg/mL of Abiraterone was injected in to the UPLC system and the percentage of assay was calculated from the resultant chromatogram.
Forced degradation studies13
Stress testing was suggested by ICH guidelines as a means to determine the intrinsic stability of drug substances. Acid degradation (2 N HCl, refluxed at 60 °C for 30 minutes), alkali degradation (2 N NaOH, refluxed at 60 °C for 30 minutes), peroxide degradation (20% hydrogen peroxide heated at 60 °C for 30 minutes), thermal degradation (samples arranged in a hot air oven at 105 °C for 6 hour), photolytic degradation (sample in an ultraviolet chamber for seven days), and hydrolysis were all applied to the solution of the standard in these investigations. The peak areas of the so-stressed samples were determined and compared to the standard's peak areas.
RESULTS AND DISCUSSION:
A new stability indicating RP-UPLC technique has been developed and validated for the simultaneous assay of Niraparib and Abiraterone using Acquity UPLC HSS T3 column and a mixture of 0.1% TFA and Acetonitrile (55:45, v/v) as mobile phase (Detection wavelength 215 nm) and the run time was 3 mins. The overlay UV absorption spectrum of Niraparib and Abiraterone was shown in Figure 2 and the chromatograms were shown in Figure 3.
Figure 2: Overlay absorption spectrum of Niraparib and Abiraterone
Method validation
Linearity, Precision, Accuracy, Robustness
Niraparib has shown linearity over the concentration range 2.5-15 μg/mL whereas Abiraterone for 12.5-75 μg/mL (Table 1) and the regression equations were found to be y = 68577x + 10960 and y = 66848x + 7064.7 respectively and the resultant calibration curves were shown in Figure 4. The LOD values for Niraparib and Abiraterone were 0.32 μg/mL and 0.46 μg/mL, respectively and the LOQ values for Niraparib and Abiraterone were 0.98 μg/mL and 1.38 μg/mL respectively. The % RSD is found to be 0.14-0.95 (Niraparib) and 0.03-1.11 (Abiraterone) in accuracy study (Table 2); 0.5-0.9 (Niraparib) and 0.5-0.6 (Abiraterone) in precision study (Table 3); 0.3-1.2 (Niraparib) and 0.4-1.3 (Abiraterone) in robustness study (Table 4) which is less than 2.0 indicating that the proposed method is precise, accurate and robust.
Table 1: Linearity study
|
Sample |
Niraparib |
Abiraterone |
||
|
|
Conc. (μg/mL) |
*Peak area |
Conc. (μg/mL) |
*Peak area |
|
1 |
2.5 |
179725 |
12.5 |
832472 |
|
2 |
5 |
351638 |
25 |
1651938 |
|
3 |
7.5 |
537291 |
37.5 |
2537291 |
|
4 |
10 |
692328 |
50 |
3413528 |
|
5 |
12.5 |
863181 |
62.5 |
4142781 |
|
6 |
15 |
1041889 |
75 |
5011889 |
|
Slope |
68577 |
66848 |
||
|
Intercept |
10960 |
7064.7 |
||
|
R2 |
0.9996 |
0.9994 |
||
*Mean of three replicates
|
A) Blank |
B) Placebo |
Figure 3: Representative chromatograms of A) Blank B) Placebo C) Niraparib and Abiraterone (Optimized)
Table 3: Accuracy study
|
Level |
Peak area |
% Recovery* (% RSD) |
||
|
|
Niraparib |
Abiraterone |
Niraparib |
Abiraterone |
|
1033378 1034231 1033501 |
5068378 5044231 5033501 |
99.2 (0.14) |
100.2 (1.06) |
|
|
100% |
1375287 1386063 1384051 |
6745287 6746063 6744051 |
100.6 (0.83) |
100.59 (0.03) |
|
150% |
1736171 1724688 1716674 |
8356171 8364688 8456674 |
100.6 (0.95) |
99.76 (1.11) |
*Mean of three replicates
Table 2: Precision study
|
S. No. |
Repeatability |
Intermediate precision (Method precision) |
||
|
Peak area |
||||
|
Niraparib |
Abiraterone |
Niraparib |
Abiraterone |
|
|
1 |
694435 |
3368723 |
683716 |
3367112 |
|
2 |
685037 |
3359146 |
692919 |
3397160 |
|
3 |
682726 |
3329550 |
683635 |
3338850 |
|
4 |
695762 |
3348758 |
684353 |
3376767 |
|
5 |
696312 |
3357001 |
684392 |
3369568 |
|
6 |
686456 |
3329675 |
683213 |
3356142 |
|
Mean |
690121 |
3348809 |
685371 |
3367600 |
|
SD |
6045.3 |
16171.3 |
3725.0 |
19594.8 |
|
% RSD |
0.9 |
0.5 |
0.5 |
0.6 |
|
A) Niraparib |
B) Abiraterone |
|
Figure 4: Calibration curves of A) Niraparib and B) Abiraterone |
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Table 4: Robustness study
|
Niraparib |
||||||
|
Condition |
Flow rate 0.3 ml/min |
Flow rate 0.4 ml/min |
MP 50:50 (v/v) |
MP 60:40 (v/v) |
Temp 230C |
Temp 330C |
|
Peak area |
686189 |
670701 |
673215 |
673735 |
68371 |
686539 |
|
696007 |
676881 |
671398 |
671770 |
687045 |
688263 |
|
|
686563 |
673669 |
683220 |
680613 |
673389 |
675898 |
|
|
699561 |
672854 |
670753 |
689799 |
690042 |
684063 |
|
|
696329 |
673055 |
670790 |
689042 |
679263 |
683654 |
|
|
Mean |
692930 |
673432 |
673875 |
680992 |
682688 |
683683 |
|
SD |
6143.5 |
2229.7 |
5318.9 |
8369.9 |
6563.0 |
4740.4 |
|
% RSD |
0.9 |
0.3 |
0.8 |
1.2 |
1.0 |
0.7 |
|
Abiraterone |
||||||
|
Peak area |
3457739 |
3461723 |
3432946 |
3384486 |
3551733 |
3395403 |
|
3393640 |
3495247 |
3420197 |
3350194 |
3558074 |
3382638 |
|
|
3385570 |
3451554 |
3419871 |
3356474 |
3546524 |
3447182 |
|
|
3454234 |
3459855 |
3450687 |
3375212 |
3451425 |
3457010 |
|
|
3451726 |
3455053 |
3453563 |
3359154 |
3556335 |
3449751 |
|
|
Mean |
3428582 |
3464686 |
3435453 |
3365104 |
3532818 |
3426397 |
|
SD |
35758.8 |
17545.2 |
16139.1 |
14230.4 |
45720.3 |
34605.1 |
|
% RSD |
1.0 |
0.5 |
0.5 |
0.4 |
1.3 |
1.0 |
Assay and Forced degradation studies of Niraparib and Abiraterone
The percentage purity of Niraparib was found to be 99.52% and that of Abiraterone 99.60%. The results of the forced degradation studies were shown in Table 5 and the respective chromatograms obtained during the forced degradation studies were shown in Figure 5. The theoretical plates were found to be greater than 2000, tailing factor is less than 1.5 and resolution is greater than 2.0 in the entire chromatographic study.
Table 5: Forced degradation studies
|
Degradation condition |
% Drug degraded* |
% Drug recovery* |
||
|
Niraparib |
Abiraterone |
Niraparib |
Abiraterone |
|
|
Acid degradation |
8.58 |
6.69 |
91.42 |
93.31 |
|
Alkali degradation |
1.50 |
1.27 |
98.50 |
98.73 |
|
Peroxide degradation |
7.35 |
7.52 |
92.65 |
92.48 |
|
Thermal degradation |
1.82 |
1.83 |
98.18 |
98.17 |
|
Photolytic degradation |
1.31 |
0.62 |
98.69 |
99.38 |
|
Hydrolysis degradation |
0.32 |
0.62 |
99.68 |
99.38 |
*Mean of three replicates
|
Acid degradation |
Alkali degradation |
|
Hydrolysis degradation |
Peroxide degradation |
|
Thermal degradation |
Photolytic degradation |
|
Figure 5: Typical chromatograms of Niraparib and Abiraterone during the forced degradation studies |
|
CONCLUSION:
The RP-UPLC technique has been developed for the quantification of Niraparib and Abiraterone in pharmaceutical dosage forms and the approach was deemed satisfactory and may be effectively utilized in normal laboratory analysis to simultaneously estimate Niraparib and Abiraterone in both bulk and pharmaceutical dosage forms.
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Received on 06.08.2024 Revised on 15.10.2024 Accepted on 02.11.2024 Published on 24.12.2024 Available online from December 27, 2024 Research J. Pharmacy and Technology. 2024;17(12):5885-5889. DOI: 10.52711/0974-360X.2024.00893 © RJPT All right reserved
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