INTRODUCTION:

Chemically Ibrutinib is 1-[(3R)-3-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl]prop-2-en-1-one. It is also be spelled as "Imbruvica".^1,4 It is available in the Indian market as tablet dosage form in the brand name as Imbruvica.⁵It is used therapeutically in chronic lymphocytic leukemia and mantle celllymphoma.⁶Ibrutinib is highly soluble in ethanol, DMSO, DMF.

IBRUTINIB

Ibrutinib is sparingly soluble in aqueous buffers. It is having Melting range of 149-158⁰C. Mechanism of action Ibrutinib is a selective and covalent inhibitor of the enzyme Bruton’s tyrosine kinase. Literature survey reveals that there is no spectroscopic and colorimetric methods available to estimate Ibrutinib either for bulk

drug or for the pharmaceutical dosage forms. The present investigation was proposed to estimate Ibrutinib in bulk and pharmaceutical dosage form by spectroscopic and colorimetric methods which were rather simple, sensitive and may prove to be cost-effective. The present work was to develop and validate as per ICH guidelines so the methods were developed and validated according to ICH guidelines for accuracy, precision and reproductively, repeatability, robustness and ruggedness.⁷

MATERIALS AND REAGENTS:

A gift sample of Ibrutinib from Pharmacylics Ltd, Mumbai used as a standard drugs. Other chemicals like Ethanol were bought from S.D fine chemicals, India.

Apparatus and equipment’s required for UV spectrophotometric and colorimetry methods:

UV-Vis double beam spectrophotometer (Model; Shimadzu:1700S, Japan), Electric Sonicator, Volumetric flasks (10ml, 50ml, 100ml), Calibrated analytical pipettes, Electronic digital balance (Techno, Mumbai)

Preparation of concentration range and standard calibration curve0:

Determination of concentration range which obeys the Lambert and Beer’s law in necessary for accuracy and reproducibility in Spectrophotometric analysis for quantitative determination of any drug. For this; Ibrutinib stock solution (100µg/ml) was prepared using pure drug in ethanol. Further dilutions were made using 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, 1.2ml, 1.4ml, 1.6ml and 1.8ml of above solutions were transferred to a series of 10ml volumetric flasks this gave a series of concentrations ranging from 2 to 18µg/ml of Ibrutinib. The final volume was made up to 10ml mark using ethanol, sonicated for 5 minutes. The resultant solutions absorbances were measured using a double beam uv-vis-spectrophotometer at wavelength of 248nm against a reagent blank. A calibration curve was plotted with concentration against absorbance. From the graph it was clear that Beer’s law was obeyed in concentration range of 2-14µg/ml and deviation was observed above these concentrations.

Estimation of Ibrutinib in tablet dosage forms:

Twenty tablets were weighed accurately and powdered. The Tablet powder equivalent to100mg of Ibrutinib was transferred into a 100ml volumetric flask and dissolved in little quantity of Ethanol. Then the solution was sonicated for 30 minutes and filtered using Whatman filter paper No#41. The filtrate so obtained was diluted with ethanol to produce100ml. Further dilutions were made with ethanol to get required concentrations within Beer’s- Lambert limits. The resultant absorbances were measured at wave length of 248nm against a reagent blank.

Reaction withN-1-Napthylethylene di amine dihydrochloride reagent for colorimetric method:

0.5N HCl, 1% NaNO₂ solution, 5% Ammonium Sulphamate, 0.2% Bratton Marshall Reagent, Ibrutinib 25µg/ml solution, Ethanol.

Sl.no

Stock solution used

0.5N HCl added

(ml)

NaNO₂added

(ml)

5% Ammonium Sulphamate

(ml)

BMR solution added

Ethanol

0ml

2ml

4ml

6ml

8ml

10ml

12ml

14ml

16ml

18ml

5ml

30ml

28ml

26ml

24ml

22ml

20ml

18ml

16ml

14ml

12ml

Sl. No

Volume of solution taken in (ml)

Conc of stock soln (μg/ml)

Absorbance

(552nm)

0ml

2ml

4ml

6ml

8ml

10ml

12ml

14ml

16ml

18ml

0 µg/ml

1 µg/ml

2µg/ml

3µg/ml

4µg/ml

5µg/ml

6µg/ml

7 µg/ml

8µg/ml

9 µg/ml

0.000

0.108

0.205

0.310

0.400

0.510

0.470

0.410

0.380

0.310

Preparation of concentration range and standard calibration curve:

Determination of concentration range which obeys the Lambert and Beer’s law is necessary for accuracy and reproducibility in spectrophotometric analysis for quantitative determination of any drug. For this; Ibrutinib stock solution (50mg/100ml) stock 1 solution was prepared using pure drug in ethanol. From stock 1 solution, 5ml was pipetted out and diluted to 100ml with ethanol for stock 2 solution. From stock 2 solution, 2ml, 4ml, 6ml, 8ml, 10ml, 12ml, 14ml, 16ml, 18ml of above solutions were transferred to a series of 50ml volumetric flasks this gave a series of concentrations ranging from 1-9 µg/ml of Ibrutinib. To each volumetric flasks, 5ml of 0.5N HCl was added, cooled in ice, 5ml of 1% ice cold NaNO₂solution was added kept for 5 minutes in ice bath. Then 5ml of 5% Ammonium Sulphamate solution was added for neutralizing excess HNO₂. Now 5ml of 0.2% BMR reagent was added and finally volumes were made up to 50ml with ethanol. The absorbances of resultant Pink colour solutions were measured at wave length of 552nm against a reagent blank. A calibration curve was plotted with concentration against absorbances (seefigure1and table1). From the graph it was clear that Beer’s law was obeyed in concentration range of 1-5µg/ml and deviation was observed above these concentrations.

Estimation of Ibrutinib in tablet dosage form:

Twenty tablets were weighed accurately and powdered. The Tablet powder equivalent to 50mg of Ibrutinib was transferred into a 100 ml volumetric flask and dissolved in little quantity of ethanol. Then the solution was mixed well and filtered using Whatman filter paper No#41. The filtrate so obtained was further diluted with ethanol to get a required concentration within Beer’s - Lambert limits. The solution after appropriate dilution was treated with 5ml of 0.5N HCl,5ml of 1% NaNO_2,5ml of 1% Ammonium Sulphamate.

RESULTS AND DISCUSSION:

Showing Beer’s range for Ibrutinib:

Sl. No.	Concentration in µg/ml	**Absorbance at λmax 248nm
1.	0	0.000
2.	2	0.085
3.	4	0.180
4.	6	0.270
5.	8	0.375
6.	10	0.470
7.	12	0.550
8.	14	0.640
9.	16	0.820
10.	18	1.000

Showing Beer’s limit for Ibrutinib pure drug:

Showing absorbance of Ibrutinib at various concentrations:

Sl. No.	Concentration in µg/ml	**Absorbance at λmax 248nm
1.	0	0.000
2.	2	0.085
3.	4	0.180
4.	6	0.270
5.	8	0.375
6.	10	0.470
7.	12	0.550
8.	14	0.640

Showing standard calibration curve for Ibrutinib:

Standard Calibration Curve for Ibrutinib:

Showing absorption of drug from tablet dosage form:

Volumeof stock Solution Used	Amount of drug (label claim) (µg/ml)	Absorbance at 248nm	Amount of drug found (µg/ml)	Percentage purity found ±S.D (%w/w)**
0.2ml	2	0.1093	2.019	100.75±0.16
0.6ml	6	0.317	6.043	100.68±1.47
1.0ml	10	0.514	9.985	99.85±0.66
1.4ml	14	0.698	13.984	99.90±0.51

(**average of three determinations)

Accuracy results for Ibrutinib:

Brands	Initial amount (µg/ml)	Amount of pure drug added(µg/ml)	Amount recovered (µg/ml)	%Recovery ±S.D**
	10	8(80%)	8.031	100.37±0.149
IMBRUVICA	10	10(100%)	9.901	99.01±0.231
	10	12(120%)	12.030	100.25±0.421

(**Average of six determinations, n=6)

Precision results for Ibrutinib:

Sl.	Conc. In (µg/ml)	Inter-day Absorbance Mean±S.D**	%C.V	Intra-day absorbance Mean±S.D**	%C.V
1.	8	0.417±0.009	0.42	0.416±0.021	0.69
2.	10	0.516±0.030	0.41	0.516±0.025	0.30
3.	12	0.609±0.023	0.67	0.612±0.031	0.31

**Average of three determinations, n=3

Showing Ruggedness parameters:

Parameters

Laboratory

Name of the instrument

Manufacturer of the chemicals used

Lab.1withanalystI

M.M.U. College Of Pharmacy, Ramanagara

Shimadzu-(model:1700S,

Japan) double beam uv-vis spectrophotometer

S.D Fine chemicals, Mumbai.

Lab.2withanalystII

Dr. H.L.T. College Of Pharmacy, Kengal, Channapatna

Systronic UV-Vis Double beam spectrophotometer

Loba chemicals, Mumbai

Showing Ruggedness results for Ibrutinib:

Sl.

Brand

Label claim

(mg)

Lab.1*with

Analyst I

Lab.2*with

analyst II

Amount found

(mg)

Recovery

± S.D**

Amount found

(mg)

% Recovery

± S.D**

Imbruvica

10.09

100.9

0.749

9.97

99.7

0.349

Showing Robustness results for Ibrutinib:

Type

Sl.

No.

Conc.

(µg/ml)

Change in temperature

Change in P^H

+5⁰C

-5⁰C

2drops

0.1NNaOH

2drops

of 0.1NHCl

Absorbance at 248nm Mean±S.D**

Pure

0.411

0.032

0.412

0.022

0.410

0.017

0.412

0.017

Drug

0.521

0.028

0.519

0.023

0.520

0.016

0.521

0.033

0.615

0.028

0.616

0.032

0.615

0.055

0.614

0.054

**Average of three determinations, n=3

Showing calibration data for Ibrutinib at 248nm:

Parameters	Calibration data at 248nm
λmax	248nm
Beer’s law limit(µg/ml)	2-14µg/ml
Molar Absorptivity	1.1161Lmol^-1cm^-1
Regression Equation(Y=a+bc)	Y=0.055X+0.016
Slope(b)	0.01426to0.01472
Intercept(a)	-0.004520to0.0016757
Correlation Coefficient(R²)	0.998
Limit of detection (LOD)	1.226µg/ml
Limit of quantitation (LOQ)	5.226µg/ml

Showing Beer’s range for Ibrutinib:

Sl. No.	Concentration in µg/ml	**Absorbance at λmax 552nm
1.	0	0.000
2.	1	0.108
3.	2	0.205
4.	3	0.310
5.	4	0.400
6.	5	0.510
7.	6	0.470
8.	7	0.410
9.	8	0.380
10.	9	0.310

Showing absorption of drug from its dosage form:

Volume of stock solution used	Amount of drug (label claim) (μg/ml)	Absorbance at 552nm	Amount of drug found (μg/ml)	Percentage purity found (%w/w)
2ml	1	0.0200	4.90	98.1%
4ml	2	0.0530	10.01	98.5%
6ml	3	0.0840	14.90	98.1%
8ml	4	0.1120	20.00	99.1%
10ml	5	0.1400	24.91	99.6%

(**average of three determinations)

Accuracy results for Ibrutinib:

Brands	Initial amount (μg/ml)	Amount of pure drug added(μg/ml)	Amount recovered (μg/ml)	%recovery ±S.D**
Imbruvica	10	16(80%)	15.00	98.02±0.114
	10	20(100%)	19.18	97.06±0.304
	10	24(120%)	23.13	98.60±0.600

(**Average of six determinations)

Showing Ruggedness results for Ibrutinib:

Sl. No

Brand

Label claim

(mg)

Lab.1*with

analyst I

Lab.2*with

analyst II

Amount found

(mg)

Recovery

± S.D**

Amount found

(mg)

Recovery

±S.D**

Imbruvica

10.09

100.9±

0.749

9.97

99.7±

0.349

Showing Robustness results for Ibrutinib:

Type

Sl.

No.

Conc.

(µg/ml)

Change in temperature

Change in P^H

+5⁰C

-5⁰C

2drops

0.1NNaOH

2drops

of 0.1NHCl

Absorbance at 248nm Mean±S.D**

Pure

Drug

0.0200±

0.022

0.113

0.022

0.004

0.402

0.013

0.1130±

0.018

0.198±0.013

0.450

0.008

0.500

0.020

0.1900±

0.018

0.160

0.022

0.415

0.035

0.600

0.040

Showing calibration data for Ibrutinib at 552nm:

Parameters	Calibration data at 552nm
Λ max	552nm
Beer’s law limit(µg/ml)	1-5µg/ml
Regression Equation(Y=a+bc)	Y=0.003X+0.001
Slope(b)	0.004670to0.10459
Intercept(a)	-0.002498to0.01364
Correlation Coefficient(R²)	0.996
Limit of detection(LOD)	1.000µg/ml
Limit of quantitation(LOQ)	2.760µg/ml

UV spectrophotometric

Determination of Beer’s limit:

The Beer’s limit felled in the range of 2-14µg/ml under given experimental condition.

Determination of absorption maxima:

The optimum wavelength was found to be 248nm

Assay:

The amount of drug found in the range of 99.85–100.75%w/w

Method validation:

The proposed method was validated in accordance to ICH guidelines.

Accuracy:

99.01– 100.37%w/w.

Precision:

The percent coefficient of variations (% C.V) was between 0.42-0.67 for intra-day and 0.30-0.69 for inter-day absorbencies.

Repeatability:

Results was not more than 0.5%.

Reproducibility:

The result (in terms of %RSD) of six determinations indicated that there were no significant variations in the data.

Robustness:

The data so obtained showed no significant variation in the absorption pattern.

Limit of detection and limit of quantitation 1.226µg/ml and 5.226µg/ml

Colorimetric Method:

Development of colour chromogen:

Finally a Pink colour complex was formed.

Beer’s limit:

1-5µg/ml

Determination of absorption maxima:

The optimum wavelength was found to be 552nm

Assay:

The amount of drug found in the range of 99.1-99.6%w/w

Method validation:

The proposed method was validated in accordance to ICH guidelines.

Accuracy:

The range of 98.02–97.06% and 98.60% w/w respectively.

Precision:

0.25-0.46 for intra-day and 0.52-0.80 for inter day absorbances.

Repeatability:

The results was not more than 0.5%.

Reproducibility:

No significant difference was observed in the result of analysis.

Ruggedness:

No significant variations in the data.

Robustness:

The data so obtained showed no significant variation in the absorption pattern of chromogen.

Limit of detection and limit of quantitation:

1.000 and 2.760µg/ml respectively.

CONCLUSION:

A new spectrophotometric method was developed to estimate Ibrutinib in pure and tablet dosage forms. Ethanolic solutions of Ibrutinib was estimated by using UV-spectrophotometer (Shimadzu 1700S, Japan) with matched 1cm quartz cells. It showed maximum absorbances at the range of 2-14µg/ml at 248nm with coefficient of correlation (R²) of 0.998. A new colorimetric method was developed to estimate Ibrutinib in pure and tablet dosage forms. Ibrutinib contains free primary aromatic amino group. Hence it can be calorimetrically estimated by Diazotization followed by coupling reaction with BMR reagent. It showed maximum absorbances at the range of 1-5µg/ml at 552nmwhich shows pink colour with coefficient of correlation (R2) of 0.996.

SUMMARY:

Ibrutinib is a newer Antitumor drug available in tablet dosage form. The data so obtained revealed that the developed method was found to be rapid, simple, accurate, method for quantitative estimation of Ibrutinib in bulk and pharmaceutical dosage form. Literature survey reveals that a few analytical methods are reported which include liquid chromatography, RP HPLC and HPTLC methods, first order derivative spectrometric determination, Liquid chromatography coupled with electro spray ionization tandem mass spectrometry, Stability indicating RP-HPLC estimation of Based on above findings, it can be said that the proposed spectrophotometric method of determination of Ibrutinib in bulk and tablet dosage form can be successfully employed in quantitative determination in bulk, finished products and in biological fluids. Ibrutinib contains free primary aromatic amino group. Hence it can be colorimetrically estimated by Diazotation followed by coupling reaction with BMR reagent which shows pink colour. The colour so produced was quantitatively measured using double beam uv-visible spectrophotometer at 552nm. The present investigation was proposed to estimate Ibrutinib in bulk and tablet dosage form by colorimetric method. The data so obtained revealed that the methods developed by the spectroscopic and colorimetric methods was found to be rapid, simple, accurate, method for quantitative estimation of Ibrutinib in bulk and pharmaceutical dosage forms. Based on the results obtained, the spectroscopic method was better than the colorimetric method.

REFERENCES:

1. A.L Honig berg, Smith, A.M., Sirisawad, M., et al. Proc. Natl. Acad. Sci. USA107(29), 13075-13080 (2010).

2. M.E.S Herman, AL Gordon, E Hertlein, et al. Blood117(23), 6287-6296(2011).

3. MWu A, Akinleye and Zhu, Hemato lJX Oncol.6, 36(2013)

4. https://en.m.wikipedia.org/wiki/Ibrutinib

5. https://go.drugbank.com/drugs/DB09053

6. Santhosillendula, Priyanka D, Shirisha V, Rao V N K, Dutt R new simple method development and validation of a Ibrutinib in bulk and pharmaceutical dosage for by RP-HPLC. International Journal of Pharmaceutical and Biological Sciences 2019; 9(1): 36-46

7. Uppala V, Tabassum A, Lavanya K, Neerja V, Sharma JVC. Method development and validation of ibrutinib by RP-HPLC in bulk and pharmaceutical dosage form. World J Pharm and Pharm Sci; 2016,5(5): 868-874.

8. National Cancer Institute. Chronic lymphocytic leukemia treatment; 2019.

9. Byrd JC, Furman RR, Coutre SE, Flinn JW, Kristie AB. Targeting BTK with ibrutinib in relapsed chronic lymphocytic leukemia; N Engl JMed2013; 369(1), 32-42.

10. Shaywitz, David. The wild story behind a promising experimental Cancer Drug; 2013.

11. Agarwal MB, Bhurani D, Shah C, Sood N, Singhal M, Kamat A, Chezhian S, Mishra S, Nagrale D. Efficacy and safety of ibrutinib in Indian patients with relapsed or refractory chronic lymphocytic leukemia and mantle cell lymphoma: cases from a named patient program. Indian J Med Pediatr Oncol 2017; 38(4): 508-515.

12. https://pubchem.ncbi.nlm.nih.gov/compound/Ibrutinib#section=Dissociation-Constants

13. https://reference.medscape.com/drug/imbruvica-ibrutinib-999896

14. Instrumental Analysis, Skoog, fifth edition, page no.312-316

15. Instrumental Methods of Chemical Analysis, Chatwal. R. Gpageno 2.116-2.122

16. Elementary Organic analysis, Principles and Chemical application, Sharma R.Y, page no 12-14

17. A Text Book of Pharmaceutical Analysis, V. Akasturi, vol 310thed., 169-81

18. https://pubchem.ncbi.nlm.nih.gov/compound/Ibrutinib

19. Owen T, Text Book of Fundamentals of UV-Visible spectroscopy1-139

20. http://www.cem.msu.edu/~reusch/virtualtext/spectrpy/UVVis/uvspec.htm#uv1

21. U.S. Pharmacopoeia (USP) general chapter $1226%, verification of compendial procedures, USP, Rockville, MD

22. U.S Pharmacopoeia (USP) general chapter $1225%, validation Of compendia procedures, USP, Rockville, MD

23. International Conference on Harmonization (ICH) (1999, Oct), harmonized tripartite guidelines Q6A specifications: Test procedures and acceptance criteria for new drug substances and new drug products: Chemical Substances.

24. International Conference on Harmonization (ICH) (2000, Nov), Harmonized Tripartite guidelines Q7A GMP for active Pharmaceutical Ingredient.

25. International Conference on Harmonization (ICH) (2005, Nov), Harmonized Tripartite Guidelines Q2(R1), Validation of Analytical Procedures; Text and Methodology.

26. International Conference on Harmonization (ICH) (2006, Sept), guidance for industry: Quality Systems Approach to Pharmaceuticals cGMP.

27. Chung Chow Chan; analytical method validation; principles and practices; Azopharma Contract pharmaceutical services, Miramar, Florida.

28. Code of Federal Regulations (CFR,), Part 211, current good manufacturing practice for finished pharmaceuticals.

29. Chann, C.C et.al (2004), analytical method validation and instrument performance verification, Jwiley, Hoboken, NJ.

30. Byrd JC, Brown JR, O’Brien S, Barrientos JC. Ibrutinib versus of atumumabin previously treated chronic lymphoid leukemia. N Engl JMed2014;371,213-223.

31. Muneer S, Ahad HA, Bonnoth CK. Novel stability indicating analytical development and validation of RP-HPLC assay method for quantification of Ibrutinibin bulk and its formulation. J Pharm Res 2017; 11(6), 712-718.

32. Bindu GH, Annapurna MM. a sensitive stability indicating RP-HPLC method for the determination of ibrutinib-Ananti-Cancer drug. Res J Pharm and Tech 2018; 11(10):4581-4591.

33. Kim ES, Dhillon S. Ibrutinib: are view of its use in patients with mantle cell lymphomaorchronic lymphocytic leukemia. Drugs. Pub Med. 75(7); 2015:769-776.

34. Parmar S, Patel K, Ibarz JP. Ibrutinib (Imbruvica): An ovel targeted therapy for chronic lymphocytic leukemia. Pharm and Therapeutics 2014; 39(7):483-87.

35. Herman SEM, Nieman CU, Farooqui M, Jones J, Mustafa RZ, Lipsky A, Saba N, Martyr S, Soto S, Valdez J, Gyamfi JA, Maric I, Calvo KR, Pederson LB, Geisder CH, Liu D, Marti GE, Aue G, Wiestner A. Ibrutinib-induced lymphocytosis in patients with chronic lymphocytic leukemia: correlative analysis from a phase II study. Macmillan Publishers Ltd. Leukemia. 2014; 28:2188-96.

36. Meloun. M, Mikesova. V, Pekarek. T, Javurek. M. The thermodynamic overlapping Pka of the antitumor drug Ibrutinib using multiwavelength UV/Visible spectroscopy and potentiometry. Journal of Analytical and Pharmaceutical Research 2(6); 2017:2-10

37. Ezzddin. E, Iqbal. M, Herqash. N.R, Elnahhas. T. Simultaneous quantitative determination of seven novel tyrosine kinase inhibitors in plasma by a validated UPLC-MS/MS method and its application to human microsomal metabolic stability study. Journal Of Chromatography. B26 October 2019 1-8

38. Rood. M.J.J, Sparidans. W.R. Bioanalysis of Ibrutinib, and its dihydrodiol and glutathionecy clemetabolites by liquid chromatography-tandem mass spectrometry. Journal of Chromatography. B2018:14-21

39. Mehta. L, Mukherjee. DLC and LC-MS/MS studies for identification and characterization of new degradation products of ibrutinib and elucidation of their degradation pathway. Journal of Pharmaceutical and Biomedical Analysis.

40. Yasu. T, Momo. K, Yasui. H, Kuroda. S. Simple determination of plasma ibrutinib concentration using high-performance Liquid Chromatography. Biomedical Chromatography.33(3)2019

41. Veeraraghavan. S, Rangasamy. M. Simultaneous qualification of lenalidomide, ibrutinib and its active metabolite PCI-45227 in rat plasma by LC-MS/MS: application to a pharmacokinetic study. Journal of Pharmaceutical and Biomedical Analysis107(2015)151-158

42. Ping. D, Guan. Y, Zhuoling. A, Pengfei. L, Lihong. LA selective and robust uplc-MS/MS method for the simultaneous quantitative determination of anlotinib, ceritinib and ibrutinib in rat plasma and its application to a pharmacokinetic study. Royal Society of Chemistry.144(18), 2019:5462-5471

43. Beauvais. D, Goossens. F.J, Boyle E, Allal. B, Lafont. T, Chatelut. E, Herbaux. C, Morschhauser. F, Genay. S, Odour. P. Development and validation of an UHPLC-MS/MS method for simultaneous quantification of ibrutinib and its dihydrodiol-metabolite in human cerebrospinal fluid. Journal of chromatography B-Analytical Technologies in the Biomedical and Life Sciences. (2018)1093-1094

44. Bende. G, Kollipara. S, Sekar. V, Saha. RUV-spectrophotometric determination of imatinib mesylate and its application in solubility studies. Original Studies. 63(2008) 641-645

45. P. Ajithkumar, A. Smith. A. Development of analytical method for imatinib mesylate by Ultraviolet spectroscopy. Asian Journal of Pharmaceutical and Clinical Research. 1(13):2020; 180-183

46. Haque. M, Bakhshi. V, Surekha. D, Gmuit. B, Reddy. P.H. Development of UV-spectrophotometric method for the determination of imatinib mesylate (ITM) in bulk and formulation. International Journal of Pharma Research and Health Sciences. 4(2):2016; 1130-1135

47. Roth. O, Varoquaux. S.O, Bouchet. S, Rousselot. P, Cstaigne. S, Rigaudeau. S, Ragguenear. V, Therond. P, Devillier. P, Molimard. M, Maneglier. B. Imatinib assay by HPLC with photodiode array UV detection in plasma from patients with chronicmyeloid leukemia. Clinica Chimicaacta 411(2010)140-146

48. Bianchi. F, Savi. M. Development and validation of a high performance liquid chromatography-tandem mass spectrometry method for the determination of imatinib in rat tissues. Journal of Pharmaceutical and Biomedical Analysis. 73(2013); 103-107

49. Madur. S, Matole. V, Kalshetti. M. UV Visible spectrophotometric method development and validation of dasatinib in bulk and solid dosage form. International Journal of Current Pharmaceutical Research. 4(12):2020; 90-93

50. Jesus SH.C, Diculescu. C.V. Redox mechanism, spectrophotometrical characterization and voltammetric determination in serum samples of kinases inhibitors and anticancer drug dasatinib.752(2015);47-53

51. Bhole. R, Bonde. C, Biradar. P. Development and validation of stability indicating HPTLC method for estimation of dasatinib and characterization of degradation products by using mass spectroscopy. Eurasian Journal of Analytical Chemistry 13(4):2018; 1306-3057

52. Ramachandra. B, Naidu. N.V.S, Sekhar. K.C. Validation of RP-HPLC method for estimation of dasatinibin bulk and its pharmaceutical dosage forms. International Journal of Pharmacy and Biological Sciences. 4(1):2014;61-68

Received on 08.03.2022 Modified on 07.04.2022

Asian J. Research Chem. 2022; 15(4):245-250.

DOI: 10.52711/0974-4150.2022.00044

INTRODUCTION:

Reaction withN-1-Napthylethylene di amine dihydrochloride reagent for colorimetric method:

0.5N HCl, 1% NaNO2 solution, 5% Ammonium Sulphamate, 0.2% Bratton Marshall Reagent, Ibrutinib 25µg/ml solution, Ethanol.

Preparation of concentration range and standard calibration curve:

Estimation of Ibrutinib in tablet dosage form:

Showing Beer’s range for Ibrutinib:

Showing Beer’s limit for Ibrutinib pure drug:

Standard Calibration Curve for Ibrutinib:

Showing absorption of drug from tablet dosage form:

Accuracy results for Ibrutinib:

Showing Ruggedness parameters:

Showing Robustness results for Ibrutinib:

Showing Robustness results for Ibrutinib:

Showing calibration data for Ibrutinib at 552nm:

Accuracy:

The range of 98.02–97.06% and 98.60% w/w respectively.

0.5N HCl, 1% NaNO₂ solution, 5% Ammonium Sulphamate, 0.2% Bratton Marshall Reagent, Ibrutinib 25µg/ml solution, Ethanol.