New UV-Visible Spectrophotometric Methods for the Determination of Aripiprazole in Bulk and Pharmaceutical Formulation

 

G.V.H. Raju*, S. Ganapathy, D.G. Sankar and P.Y. Naidu

Department of Pharmaceutical Sciences, Andhra University, Visakhapatnam-530003

*Corresponding Author E-mail: gadepalli_raj@yahoo.com

ABSTRACT:

 

INTRODUCTION:

Aripiprazole is chemically a quinolinone derivative, used as an anti-psychotic agent.Chemically, Aripiprazole is known as 7-[4-[4-(dichlorophenyl) piperizin-1-yl]-3, 4-dihydroquinolinone. The empirical formula is C₂₃H₂₇Cl₂N₃O₂ and its molecular weight is 448.38. It is a white crystalline powder and practically insoluble in water and its solubility is pH dependent. The chemical structure is given below ^1-6

 

It is not official in any pharmacopoeia and only few liquid chromatographic and UV Visible Spectrophotometric   procedures have been reported for the determination of Aripiprazole in Bulk and Pharmaceutical dosage forms^14-16.

 

In the present investigation simple, sensitive and reproducible UV Visible spectrophotometric methods are  developed for the determination of Aripiprazole (ARIP) in their pharmaceutical dosage forms  using Folin- Ciocalteu (FC) reagent in presence of 4% sodium Hydroxide solution, MBTH- CAS , Fecl₃ -1, 10PTL, and with DCQC(2,6-dichloroquinone N-chlorimide ,Gibbs Reagent), NQS(1,2-napthaquinone-4-sulphonic acid). In Method A the formation of colored species having λ_max  at 660nm is due to the Oxidative Coupling reaction with MBTH in Presence of  CAS. In Method B the formation of Colored species having λ_max  at 466nm is due to reaction of Reduced Ferric ion to Ferrous ion with 1,10 Phenanthorline. In method C the formation of colored species  is based on the reduction of phosphomolybdotungstic acid(FC reagent), by Aripiprazole in presence of 4% sodium Hydroxide solution, thereby producing reduced species having characteristic blue colour with maximum absorption at 680 nm. In method D the formation of colored species is based on coupling reaction of DCQC (2,6-dichloroquinone N-chlorimide, Gibbs Reagent) with the Drug to give colored species having maximum absorption at 520nm. In method E the formation of colored species is based on reaction of NQS (1,2-napthaquinone-4-sulphonic acid) with the drug by which the Sulphonate group of the napthaquinone sulphonic acid is replaced by an amino group to give N-arylamino napthaquinone having maximum absorption at 460nm. Method F represents determination of Aripiprazole  and its dosage forms in pH 3.0Buffer : Acetonitrile (62:38v/v) which is having absorption maxima at 250nm

 

EXPERIMENTAL:^7-13

Instrument: A Shimadzu UV-VIS Spectrophotometer was used for absorbance measurements

Reagents:

All the Chemicals used were of analytical grade

 

Method A:

i)       MBTH (0.2%): 200mg of MBTH was dissolved in 100ml of distilled water

ii)      CAS (1%): 1g of Cerric ammonium Sulphate (CAS) was dissolved in 0.72M Sulphuric acid

iii)     Diluent: Methanol

 

Preparation of Standard solution(100µg/ml): About 100mg of Aripiprazole(ARIP) working standard was accurately weighed and transferred into 100mL volumetric flask, 70ml of methanol is added sonicated  to dissolve and the total volume is made up to volume with diluent. Further 5ml of the resulting solution is taken into 50ml volumetric flask and made up to volume with diluents

 

Preparation of Test Solution: Twenty tablets were taken weighed and powdered. Sample equivalent to 100mg of Aripiprazole was accurately weighed and transferred in to 100ml volumetric flask, 70ml of diluent was added sonicated for 20min and made up to volume with diluent. 10 ml was taken and centrifuged at 2500rpm. Further 5ml of the resulting solution is taken into 50ml volumetric flask and made up to volume with diluent.

 

Method B:

i)       Fecl₃ (0.9%): 900mg of Fecl₃ was dissolved in 100ml of distilled water

ii)      1, 10 PTL (0.125%): 125mg of 1, 10 PTL was dissolved in 100ml of distilled water

iii)     Diluent   : Methanol

 

Preparation of Standard solution (100µg/ml): About 100mg of Aripiprazole working standard was weighed and transferred in to a 100ml volumetric flask, dissolved in 70ml of diluent and made up to volume with diluent. Further 5ml of the resulting solution is taken into 50ml volumetric flask and made up to volume with diluent.

 

Preparation of Test Solution: Twenty tablets were taken weighed and powdered. Sample equivalent to 100mg of Aripiprazole was accurately weighed and transferred in to 100ml volumetric flask, 70ml of diluent was added sonicated for 20min and made up to volume with diluent. 10 ml was taken and centrifuged at 2500rpm. Further 5ml of the resulting solution is taken into 50ml volumetric flask and made up to volume with diluent.

 

Method C:

i)       FC Reagent: 50ml of Folin - Ciocalteu reagent (2N) was diluted to 100 ml with distilled water.

ii)      NaOH (4%) : 4.0g of Sodium Hydroxide was dissolved in 100ml of distilled water

iii)     Diluent: Methanol

Preparation of Standard solution(100µg/ml): About 100mg of Aripiprazole (ARIP) working standard was accurately weighed and transferred into a 100ml volumetric flask dissolved in 70ml of diluent and made up to volume with diluent.5ml of the above stock is taken into 50ml volumetric flask and made up to volume with Diluent.

 

Preparation of Test Solution: Twenty tablets were taken weighed and powdered. Sample equivalent to 100mg of Aripiprazole was accurately weighed and transferred in to 100ml volumetric flask, 70ml of diluent was added sonicated for 20min and made up to volume with diluent. 10 ml was taken and centrifuged at 2500rpm. Further 5ml of the resulting solution is taken into 50ml volumetric flask and made up to volume with diluent.

 

Method D:

i)       DCQC solution (0.04%): 40mg of DCQC was dissolved and diluted to 100 ml with isopropyl alcohol.

ii)      Buffer Solution (pH 9.4): Mix 250ml of 4.0g of Sodium Hydroxide was dissolved in 100ml of distilled water

iii)     Diluent: Methanol

 

Preparation of Standard solution (1000µg/ml): About 100mg of Aripiprazole (ARIP) working standard was accurately weighed and transferred into a 100ml volumetric flask dissolved in 70ml of diluent and made up to volume with diluent.

 

Preparation of Test Solution: Twenty tablets were taken weighed and powdered. Sample equivalent to 100mg of Aripiprazole was accurately weighed and transferred in to 100ml volumetric flask, 70ml of diluent was added sonicated for 20min and made up to volume with diluent. 10 ml was taken and centrifuged at 2500rpm

 

Method E:

i)       NQS solution (0.5%): Prepared by dissolving 500 mg of NQS in 100 ml of distilled water

ii)      Buffer Solution (pH 8.0) : Prepared by mixing 30 ml of potassium hydrogen phosphate (0.067 M) and 770 ml of disodium hydrogen phosphate (0.067 M) and the pH of the solution was adjusted to 8.0.

iii)     Diluent: Methanol

 

Preparation of Standard solution(1000µg/ml): About 100mg of  Aripiprazole(ARIP) working standard was accurately weighed and transferred into a 100ml volumetric flask dissolved in 70ml of diluent and made up to volume with diluent.

 

Preparation of Test Solution: Twenty tablets were taken weighed and powdered. Sample equivalent to 100mg of Aripiprazole was accurately weighed and transferred in to 100ml volumetric flask, 70ml of diluent was added sonicated for 20min and made up to volume with diluent. 10 ml was taken and centrifuged at 2500rpm

 

 

Method F:

i)       Preparation of pH 3.0 Buffer solution:

Weigh and dissolve 2.725 g of potassium dihydrogen orthophosphate in 1000mL of milli-Q water in a suitable container, add 10mL of triethylamine and mix, and adjust the pH of the solution to 3.0 ± 0.05 with dilute ortho phosphoric acid.

ii)      Preparation of Diluent:

Mix 620mL of pH 3.0 Buffer with 380mL of Acetonitrile

 

Preparation of Standard solution(100µg/ml): About 100mg of Aripiprazole(ARIP) working standard was accurately weighed and transferred into a 100ml volumetric flask dissolved in 70ml of diluent and made up to volume with diluent.5ml of the above stock is taken into 50ml volumetric flask and made up to volume with Diluent.

 

Preparation of Test Solution: Twenty tablets were taken weighed and powdered. Sample equivalent to about 20mg of Aripiprazole (ARIP) was accurately weighed and transferred into a 100ml volumetric flask dissolved in 70ml of diluent and made up to volume with diluent. Few ml was taken and centrifuged at 2500rpm.5ml of the Clear Centrifuge is taken in to 50ml volumetric flask and made up to volume with diluent.

 

Assay Procedures:

For Method A Volumes of standard solution ranging from (0.1-1.0ml; 1ml=100µg) were transferred into a series of 10ml graduated volumetric flasks. A 1ml portion of CAS was added to each flask and shaken gently for 5 min. Then 2ml of MBTH solution was added to each flask and kept aside for 15min for maximum color development .Appropriate volume of diluent was added to each flask to bring the total volume 10ml.The absorbance was measured at 660nm against reagent blank. The Test solution was treated in the same manner as standard and measured the absorbance at 660nm

 

For Method B Volumes of standard solution ranging from (0.4-2ml; 1ml=100µg) were transferred into a series of 10ml graduated volumetric flasks. Then 1ml portion of FeCl₃ and 2.0ml of 1,10 PTL were added successively and set aside for 10min and the total volume made up to 10ml with methanol. The absorbances of the colored complex solutions were measured at 466nm against reagent blank prepared similarly. The Test solution was treated in the same manner as standard and measured the absorbance at 466nm.

 

For Method C 1ml portion of FC reagent was taken in a series of 10ml graduated volumetric flasks. Volumes of standard solution ranging from (1-5ml; 1ml=100µg) were transferred into a each flask and shaken gently for 5 min. Then 2ml of NaOH solution was added to each flask and kept aside for 15min for maximum color development .Appropriate volume of methanol was added to each flask to bring the total volume 10ml.The absorbance was measured at 680nm against reagent blank. The Test solution was treated in the same manner as standard and measured the absorbance at 680nm.

 

For Method D   Volumes of standard solution ranging from (0.1 – 1.2 ml, 1000 g/ml) were transferred into a series of 10ml graduated volumetric flasks, 5.0 ml of buffer pH 9.4, 2.0 ml of  DCQC solution was added. The volume in each tube was made up to 10 ml with distilled water. The absorbances were measured at 520 nm against a similar reagent blank. The Test solution was treated in the same manner as standard and measured the absorbance at 520nm.

 

For Method E Volumes of standard solution ranging from (2 – 12 ml, 1000g/ml) were transferred into a series of 10ml graduated volumetric flasks. Then 1 ml of NQS, 5.0 ml of pH 8.0 buffer were added to each flask, the volume was made up to the mark with MeOH and kept aside for 30 min at laboratory temperature. The absorbencies were measured at 460 nm against a reagent blank. The Test solution was treated in the same manner as standard and measured the absorbance at 460nm.

 

For Method F Volumes of standard and test solution ranging from (0.5-2.5ml; 1ml=100µg) were transferred into a series of 10ml graduated volumetric flasks and made up to volume with diluent. The resulting solutions were measured the absorbance at 250nm using the diluent as blank

 

The amount of Aripiprazole (ARIP) present in the sample solutions(X) using the proposed methods was calculated by formula given below and the results are given in Table-2.

 

             A_T        D_s            A_w             P

X =       ---- X ------X   ----   X ---- X 100

              A_s        D_T        L         100

A_T   = Absorbance of sample solution

A_s   = Absorbance of standard solution

D_s = Dilution factor for Standard

D_T   = Dilution factor for Test Solution

A_{w =} Average weight of sample in mg

L    = Label Claim

P =Potency of Standard on as is basis

 

RESULTS AND DISCUSSION:

The optical characteristics such as Beer’s law limits (µg/ml), Sandell’s sensitivity (µg/cm²/0.001 A.U), Correlation coefficient (r), % Relative standard deviation(Calculated from six determinations), % Range of error (0.05 confidence limits and 0.01 confidence limits), Regression equation(I+ ac) for the proposed method is calculated and summarized in table-1.The results obtained with the proposed methods for dosage forms is Compared with the results obtained with HPLC method and presented in Table-2.The Proposed method is Validated and found to be Specific, Accurate, Precise and Linear. Interference studies revealed that the common excipients used in the dosage forms do not interfere in the estimation of Aripiprazole (ARIP) using the proposed methods.

TABLE-1 OPTICAL CHARACTERISTICS AND PRECISION

Method Parameters	Method A	Method B	Method C	Method D	Method E	Method F
λmax	660nm	466nm	680nm	520nm	460nm	250nm
Beer’s law limits(µg/ml)	1-10	4-20	10-50	10-120	20-120	5-20
Sand ell’s Sensitivity (µg/cm2/0.001 A.U)	0.00881	0.02970	0.05848	0.18450	0.15221	0.03135
Regression equation(I + ac) Slope (a) Intercept (I)	0.11573 -0.00128	0.03384 0.00322	0.01749 -0.00430	0.00539 0.00805	0.00647 0.00567	0.03190 0.00470
Correlation coefficient(r)	0.999	0.999	0.999	0.999	0.999	0.999
%RSD	0.3240	0.529	0.397	0.4710	0.375	0.2260
% Range of error Confidence limits with 0.05 level Confidence limits with 0.01 level	0.271 0.401	0.442 0.654	0.332 0.491	0.394 0.583	0.314 0.464	0.189 0.280

* Mean of six determinations

 

 

TABLE-2 EVALUATION OF ARIPIPRAZOLE IN PHARMACEUTICAL PREPARATIONS

Label Claim Mg/tablet	Amount Obtained (mg)*
	Proposed method					Reference Method (UV)
	Method A	Method B	Method C	Method D	Method E
20	19.98 ±0.0440	19.97 ±0.0308	20.06 ±0.0796	20.07 ±0.096	19.969 ±0.075	19.997 ±0.045

*Mean ±SD of six determinations

 

%Recovery
Method A	Method B	Method C	Method D	Method E	Reference Method (UV)
99.90 ± 0.220	99.85 ± 0.158	100.29 ± 0.3978	100.35 ± 0.4800	99.84 ± 0.3743	99.98 ± 0.233

.** Mean ±SD of six determinations

 

The Calibration Curve Figure 1, Figure 2, Figure 3, Figure 4, Figure 5 and Figure 6 for the Proposed methods was illustrated in the Concentration range 1 to 10 µg mL^-1 of Aripiprazole(ARIP) for Method A, 4 to 20 µg mL^-1 for Method B, 10 to 50 µg mL^-1 for Method C, 10 to 120 µg mL^-1 for Method D, 20 to 120 µg mL^-1 for Method E and  5 to 20 µg mL^-1 for Method F.

Figure 1. Calibration Curve of Aripiprazole - Method A (MBTH-CAS)

 

Figure 2. Calibration Curve of Aripiprazole - Method B (FeCl₃-1,10 PTL)

 

Figure 3. Calibration Curve of Aripiprazole –Method C (FC reagent)

 

Figure 4. Calibration Curve of Aripiprazole –Method D (DCQC)

 

Figure 5. Calibration Curve of Aripiprazole –Method E (NQS Reagent)

 

Figure 6. Calibration Curve of Aripiprazole in pH 3.0 buffer: CAN (62:38)

To Check the recovery using the proposed method known amounts of pure drug was added to the Placebo used in the pharmaceutical preparation of Aripiprazole and the mixtures were analyzed by the proposed method and the %Recoveries are given in the table-2. In conclusion the proposed methods are simple, sensitive and accurate and can be used for the routine determination of Aripiprazole (ARIP) in pharmaceutical preparations.

 

ACKNOWLEDGEMENTS:

The authors wish to thank the Orchid Healthcare for providing the samples of Aripiprazole.

 

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Received on 13.05.2010        Modified on 19.05.2010

Accepted on 24.05.2010        © AJRC All right reserved