INTRODUCTION:

Alectinib hydrochloride is an orally accessible tyrosine kinase inhibitors with anticancer properties. It is described chemically as 9-ethyl-6,6-dimethyl-8-[4-(morpholin-4-yl)piperidin-1-yl]-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrile hydrochloride with molecular weight 519.08 Dalton, and its empirical formula C₃₀H₃₄N₄O₂.HCl^1-2. The chemical Structure of Alectinib Hydrochloride is depicted in (figure 1). The Food and Drug Administration granted approval to alectinib hydrochloride for the treatment of patients with anaplastic lymphoma kinase-positive, metastatic non-small cell lung cancer (NSCLC) in 2017. It works by blocking the action of an abnormal protein that signals cancer cells to multiply³. Alectinib hydrochloride is official in the Indian Pharmacopoeia, United States Pharmacopoeia and British Pharmacopoeia. Literature survey reveals few analytical methods^4-9 for the determination of alectinib hydrochloride in the pharmaceutical products as well as in the biological samples. The literature survey shows that ultraviolet spectrophotometric method is simple and cost effective compared to chromatographic methods such as liquid chromatography^10-12, Gas chromatography^13-14 and thin layer chromatography^15-16. The aim of current research work was to develop a simple, precision, accurate eco-friendly and cost effective ultraviolet spectrophotometric method for the quantification of alectinib hydrochloride in capsule dosage form using green solvent.

Figure 1: Chemical structure of alectinib hydrochloride

MATERIALS AND METHODS:

Alectinib hydrochloride was procured as gift sample for Sun Pharmaceutical Industries Ltd., Vadodara, Gujarat, India and capsules formulations with a strength of 150 mg were prepared in the laboratory using common excipients. Methanol of analytical grade was purchased from S D Fine Chemicals, Mumbai, India. The analysis of samples were carried out on UV-Visible spectrophotometer (1800, Shimadzu Corporation, Japan). An electronic balance (AX200, Shimadzu, Ltd., Japan) and ultrasonic bath (Leela Sonic- 150, Leela Electronics, India) was used in the this research work. All the instruments and glassware were calibrated prior to use.

Preparation of stock and working standard solutions

Accurately weighed 50 mg of alectinib hydrochloride and was transferred to 50 mL volumetric flask. It was dissolved and diluted with methanol to obtain a final concentration of 1000 µg/mL (stock solution). An appropriately diluted the stock solution to obtain 100 µg/mLof working standard solution.

Method validation

The developed spectrophotometric method was validated in accordance to ICH Q2 (R1) guideline¹⁷. Linearity was assessed by constructing a calibration curve over a range 2-12 µg/mL of alectinib hydrochloride. Calibration curves were developed by plotting absorbance vs. concentration (n=3) and correlation coefficient and regression equation were calculated. The recovery experiment was performed by spiking standard solutions of drug (4, 5, 6 µg/mL) into previously analysed formulation sample of 5 µg/mL at the level of 80, 100 and 120 %. In the precision study, repeatability of the method was confirmed by measuring the absorbance of 5 µg/mL solution of drug at 339 nm for six times and % RSD was calculated. Intraday precision was carried out by determining three different concentrations of alectinib hydrochloride (2, 5, 12 µg/mL) for three times on a same day and interday precision was checked by determining the same concentrations for three times on three consecutive days and % RSD was calculated. Limit of detection (LOD) and limit of quantitation (LOQ) was calculated in accordance to the equation given in ICH Q2 (R1) guideline. Specificity of alectinib hydrochloride was performed by using synthetic mixture having standard drug and commonly used excipients in the manufacturing of capsule formulation. Robustness was performed by making small deliberate changes in two internal factors - detection wavelength and solvent compositions, and results were examined. The % RSD was calculated for changes in each condition. Solution stability studies of drug sample were carried out at room temperature. The absorbance of the solutions kept under the room temperature were measured periodically and compared with initial concentration of the solution. The significant variation was observed after twelve hours at room temperature.

Assay of capsule formulation

Twenty capsules of 150 mg strength were weighed and carefully remove the content from the capsule shell. The powder quantity equivalent to 100 mg of alectinib hydrochloride was accurately weighed on electronic balance and transferred into a 100 mL volumetric flask containing 50 mL methanol, dissolved for 15 min and diluted up to the mark with same solvent. The resultant solution was mixed and filtered through Whatman filter paper No. 1. An aliquot of 1 mL from the resultant solution was transferred to 100 mL volumetric flask and diluted with methanol to obtain 10 µg/mL concentration. The absorbance of the resultant solution was measured at 339 nm using methanol as blank. The concentration of sample solution was calculated from the standard calibration curve of alectinib hydrochloride.

RESULTS ADN DISCUSSION:

The standard solution of drug (8 µg/mL) was prepared in methanol and was scanned in the range of 200 to 400 nm against methanol as solvent blank. The detection wavelength for the measurement of alectinib hydrochloride was selected at 339 nm. The zero order ultraviolet spectrum of standard drug is shown in (figure 2). Linearity of the developed method was found to be 2 – 12 µg/mL, with value of correlation coefficient (r²) of 0.9996. Accuracy of method was determined by performing recovery studies and was found in the range of 99.56 – 101.43 %, indicates the method is accurate. The precision data for the method was found within the limits indicates the method is precise. Detection limit and quantitation limit values were found to be 0.57 µg/mL and 1.71 µg/mL, indicates that the method is sensitive. The % RSD was found within the limits for robustness study indicates that the method is robust. The assay contents of drug was found to be 99.84 % indicates that the drug products complies with the labeled claims. Results of validation parameters are summarized in (Table 1).

Figure 2: Ultraviolet spectrum of alectinib hydrochloride (8 µg/mL) in methanol

Table 1: Validation parameters for alectinib hydrochloride

Parameters	Results
Detection wavelength (nm)	339
Regression equation	y = 0.069 x + 0.007
Beer’s Law limits (mg/mL)	2 - 12
Limit of detection (mg/mL)	0.57
Limit of quantification (mg/mL)	1.71
Precision (% RSD) · Repeatability · Intraday · Inter-day	0.6895 0.32 - 1.31 0.56 – 1.87
Accuracy (% Recovery)	99.56 – 101.43
Robustness	Robust
Specificity	Specific

CONCLUSION:

The satisfactory results of validation parameters and drug contents indicate that the developed method is simple, accurate, precise and robust. Hence, the proposed spectrophotometric method can be routinely used for the estimation of alectinib hydrochloride from capsule dosage form without interference of excipients in the quality control laboratory.

ACKNOWLEDGMENTS:

The authors are thankful to Sun Pharmaceutical Industries Ltd., Vadodara, Gujarat, India for providing gift sample alectinib hydrochloride.

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Received on 04.07.2023 Modified on 09.11.2023

Asian J. Pharm. Ana. 2024; 14(3):119-121.

DOI: 10.52711/2231-5675.2024.00020