Determination and Development of UV-Spectrophotometric method for the Amodiquine Hydrochloride by using quality by design (QBD) approach

 

Patil Karishma Ashok¹, Firke Sandip Dinkar²*

 

ABSTRACT:

To develop a simple, accurate, and inexpensive UV sepectrometric method for the determination of the Amodiquine hydrochloride by using quality by design (QbD) approach. The double beam spectrophotometer of Shimadzu UV-1800 by using a determination of UV spectrophotometer method. The amodiquine Hydrochloride is soluble with Distilled Water & methanol (9:1) as solvent and wavelength of 341 nm was selected as absorbance (𝜆max). The effect to check normal variables and selected variables on spectrum characteristics were studied for the selection of critical parameters and proposed method was validated for linearity, accuracy, limits of detection and quantification as per the ICH Q2 (R1) International Conference on Harmonization guidelines. The Linearity of this method was excellent on 3 to 21 µg/mL concentration range and correlation coefficient (r²) of 0.998. The (LOD) and (LOQ) were getting found to be 0.038µg/mL and 0.084 µg/mLrespectively. The amodiquine Hydrochloride recovery was found to be 99.6%.

 

 

 

INTRODUCTION:

The Malaria disease is caused by protozoan parasite which is transmitted from person to person through bites from infected plasmodium Female Anopheles Mosquitoes.

 

The chemically amodiquine hydrochloride (ADQ) is, 4-[(7-Chloro-4quinolyl) amino]-α-(diethylamino)-o-cresol dihydrochloride dihydrate; 4-[(7-chloro4-quinolinyl) amino]-2-[(diethylamino)-methyl] phenol. The literature survey revealed various analytical methods for estimation of amodiquine HCl by UV in the different dosages form. In this paper a QbD approach was used to determine ADQ.^[1,2,3]

 

Figure 1: chemical structure of amodiquine hydrochloride

 

The concept of quality by design (QbD) is used for the development of pharmaceutical processes to ensure a predefined objective of process and product quality.^[4] QbD concepts have detail study on in (ICH) guidelines International Conference on Harmonization. The ICH guideline Q8 (R1)^[5] for Quality By Design is a systematic process to development^[6] begins with predefined aim and objectives of the process and product understanding and process will be control on the base of quality risk management and sound science.^[7] QbD is a systematic approach for the development of product through an understanding of the effects of various input variables (Ex. parameters, process, materials) on the final material. Thus, QbD approach defines accurate ranges of the input parameters within which the quality of the final product is assured.^[8] In a same manner describe for analytical methods, QbD approach involves a full understanding of how the analytical technique operating conditions and attributes interferes with the analytical performance. Factors to study in analytical quality by design (AQbD) approach may include the type of analytical technique chosen, chemical & reagents used, and instrumental parameters.^[9]

 

The principles of applying QbD to analytical methods as to processes and manufacturing of product.^[10] The most of researchers have been systematically explain QbD principles to analytical process development       method.^[11-16] The AQbD can be used for determination and development of a precise and cost reliable analytical method which is applied at the any level the whole product lifecycle. The higher authorities have provided recently changing of analytical method without revalidation of the AQbD approach has been design and implemented during development of analytical method.^[17]

 

AQbD approach is a first stage to set an analytical target profile (ATP) for the method. ATP defines the goal of the analytical method development process and it is the indicators of method performance. An ICH guideline on validation of analytical procedures, ICH Q2 (R1), has given various method performance characteristics for an analytical method. Thus, a QbD based UV spectrometric method can be developed by considering the ICH guidelines Q2 (R1).

 

The goal of the present investigation was to developed a simple, robust, fast and economical ^[18] UV spectrometric method^[19,20,21] for the estimation of Amodiquine hydrochloride by using analytical quality by design (AQbD) approach. For development QbD approach to UV spectrophotometric analytical method, the effect of method input variables on spectral shape, intensity of absorbance, and absorbance maxima (𝜆max), were studied and critical parameters were selected for the proposed method. Then proposed method was validated ^[22] as per the ICH guidelines ICH Q2 (R1).

 

MATERIALS AND METHODS:

Equipment and chemicals:

UV Spectrophotometer (Shimadzu UV-1800 double beam spectro-photometer). Amodiquine hydrochloride (ADQ) was purchased from Vital Laboratories Pvt. Ltd. GIDC, Vapi-396195. Dist. Valsad Gujarat. All other chemicals used were of analytical grade reagent.

 

 

Preparation of stock solution:

The solubility of ADQ in various solvents such as distilled water, 0.01N hydrochloric acid, chloroform and methanol was determined. The maximum solubility of ADQ was found to be in Distilled Water. Hence the Distilled Water is used for the preparation of standard stock and further dilution. The stock solution was prepared by dissolving 10 mg of ADQ in 100 ml (9:1) Distilled Water and methanol to obtain 100𝜇g/ml concentration. Standard working solution of 10𝜇g/ml was prepared from the stock solution by dilution with Distilled Water and used for initial spectral scan in the UV spectrophotometer.

 

Determination of wavelength of maximum absorption:

Standard working solution (10 𝜇g/mL) of ADQ was scanned from 200-400 nm in the UV spectrophotometer for the selection and determination of analytical wavelength. ADQ showed maximum absorbance (𝜆max) at 341 nm (fig. 1). Hence 341 nm was selected as analytical wavelength.

 

Implementation of AQbD approach in the development of the analytical method:

For AQbD approach, the Ishikawa diagram was used to study the relationship between variable input parameters and the method performance characteristics of the spectrophotometric analytical methods.

 

Fig. 2: UV-spectrum of amodiquine hydrochloride showing 𝜆max at 341 nm

 

Fig 3: Ishikawa diagram showing the relationship between variable input parameters and the method performance characteristics

The selection of solvent is based on the maximum solubility of the amodiquine hydrochloride. The maximum solubility of ADQ was found to be in Distilled water hence the Distilled water was selected as a solvent for determination of UV spectrophotometer analytical method of ADQ. All the other parameters that are shown in Ishikawa diagram, the spectrum of absorbance were recorded by scanning standard working solution (10 𝜇g/mL) in that selected solvent range from 200-400 nm in the UV spectrophotometer.

 

The shape of spectra, sharpness, and the intensity of absorbance spectrum were recorded and compared at different scan speed such as very slow, slow, medium, and fast and varied interval of sampling such as 0.1, 0.2, and 0.3 nm.^[23] Thus, indicating no significant changes in shape of spectral, sharpness, and absorbance’s intensity of spectrum due to variation in sampling interval and scan speed. The environment conditions also play the important role in the stability of the solution of whole period of time.

 

On the basis of observation, the selected some critical parameters in (table 1) and by using selected variable parameters, the method is validated as per the ICH guidelines Q2 (R1).

 

Table 1: Selected critical parameter for spectrophotometric analytical method of ADQ

PARAMETER	SELECTED VARIABLES
Wavelength	341nm
Solvent	Distilled water
Scan speed	Fast
Sampling interval	0.3 nm
Environment	Time

 

VALIDATION:

The analytical method all the selected critical parameters should be within a range with performance method characteristics to achieve the analytical target profile of this method. The ICH laid down various method performance of characteristics for an analytical method in Q2 (R1) ICH guidelines. Thus, for an analytical UV spectrophotometric method, it is appropriate to determine and validate according to ICH guidelines Q2 (R1) on the selected critical parameter in order to implement AQbD approach.

 

The selected critical parameters using, developed method is further validated as per the ICH guidelines Q2 (R1).^[7] The different characteristics studied were system suitability, linearity, accuracy, and precision, limit of detection (LOD) and limit of quantification (LOQ).

 

System suitability:

System suitability is done to demonstrate the suitability of the UV spectrophotometer system being used for the analysis. Six replicates of standard solution (10 𝜇g/mL) of ADQ were prepared from a stock solution in the selected solvent (Distilled water) and absorbance was determined at 341 nm of each replicate using UV spectro-photometer. Percentage relative standard deviation (% RSD) was calculated for the absorbance.^[7]

 

Linearity:

According to the ICH guidelines, the linearity of an analytical procedure determines that the test results are directly proportional to the concentration (amount) of analyte in the sample. For linearity study, seven solutions of different concentrations (3, 6, 9, 12, 15, 18 and 21µg/mL) were prepared in Distilled water from a standard stock solution of ADQ and absorbance of each solution was noted at 341 nm in triplicate. The calibration curve was prepared by plotting the absorbance against concentration and % RSD, and the correlation coefficient was calculated by regression analysis. ^[7]

 

Accuracy:

The accuracy of an analytical procedure shows the closeness of results with the conventional true value. Accuracy was determined by recovery study of ADQ. To the known amount of standard solution (10𝜇g/mL), a known amount of standard stock solution was added at a different level, i.e. 50%, 100% and 150% to get final concentration of 15𝜇g/mL, 20𝜇g/mL, and 25𝜇g/mL. Then these solutions were reanalyzed for drug content. Triplicate set of each levels were prepared for the experiment. The recovery of sample, and % RSD were calculated.^[7]

 

Precision:

According to the ICH guidelines, the precision of an analytical procedure determines the closeness of results obtained by multiple measurements of the same homogeneous sample. Repeatability (intra-day precision) and intermediate precision (inter-day precision) were done to show the precision of the method.

 

To demonstrate repeatability (intra-day precision) of the test method, six replicates of the 10 𝜇g/mL concentration (n=6) were analyzed on the same day. % RSD of assay result of six replicates was calculated. Similarly, for intermediate precision (inter-day precision), six replicates of the 10 𝜇g/mL concentrations were analyzed for assay on three consecutive days and % RSD was calculated.^[7]

 

Limit of detection (LOD) and limit of quantification (LOQ):

Limit of detection is the lowest amount of analyte in a sample that can be detected, but not necessarily quantified as an exact value and limit of quantification is the lowest concentration of an analyte in the sample that can be determined with accuracy and precision. Limit of detection and limit of quantification concentrations for ADQ were determined based on the residual standard deviation of response and slope method as per ICH guideline. Calibration curve prepared in linearity study was used for this purpose. For LOD calculation equation (3.3 x δ)/S and for LOQ equation (10 x δ)/S was used. Where σ is the standard deviation of the response and S is the slope of the calibration curve.^[7]

 

RESULTS AND DISCUSSION:

The representative chromatogram of the standard solution of mixture is shown in Figure 1. Results were found to be linear in the concentration range of 3-21 mg/mL for ADQ. The correlation coefficients for the plots were 0.998 for ADQ. The proposed method was also evaluated.

 

System Suitability:

The absorbance of six replicate of standard solution (10𝜇g/mL) of ADQ is reported table 2. For system suitability % RSD of absorbance of replicate solutions is should not more than 2.^[24] The results obtained was meets the system suitability requirements, this indicates that the system was suitable for the analysis. ^[25]

 

Linearity:

The calibration plot of absorbance versus concentration was found to be linear over the concentration range of 3-21 µg/ml as shown in fig 4. RSD was found to have small value 0.00283% while correlation coefficient (r²) has high value 0.998 (table 3). Thus, indicating that test results were directly proportional to the concentration (amount) of analyte in the sample. ^{[26, 27, 28]}

 

Table 2: Result of system suitability studies

ADQ replicate of sol.	Abs. At 341nm
1	0.612
2	0.624
3	0.646
4	0.631
5	0.619
6	0.625
Mean	0.6262
SD	0.0116
% RSD	1.8553

 

Fig. 4: Calibration curve plot of amodiquine hydrochloride at 341 nm

Table 3: Summary of Validation parameters for Amodiquine Hydrochride

Sr. No	Parameters	Result
1	Linearity Range (µg/mL)	3-21 µg/Ml
2	Correlation coefficient (r2)	0.998
3	Precision (%C.V.) 1.Repeatability	0.165-0.285
	2.Intraday precision	0.186-0.564
	3.Interday precision	0.268-0.865
4	Accuracy (%recovery)	99.6%
5	Limit of Detection (L.O.D.) (µg/mL)	0.38 µg/mL
6	Limit of Quantification (L.O.Q.) (µg/mL)	0.084 µg/mL

 

CONCLUSION:

A simple, rapid, sensitive, accurate, precise and inexpensive spectro-photometric method was developed for estimation of amodiquine hydrochloride in bulk by using analytical quality by design” (AQbD) approach. On the basis of an investigation of the effect of method input variables on absorbance pattern, the critical parameters have been selected for proposed method and it was further validated as per the ICH guidelines. The developed method does not involve complexity thus have economic advantages over common chromatographic methods. Therefore, developed spectrophotometric method can be used flexibly and efficiently for the determination of amodiquine either in bulk or in the dosage formulations.

 

ACKNOWLEDGMENTS:

The authors are thankful to the management, Principal Dr. S.J Surana Sir and the staff of R.C.P.I.P.E.R. Shirpur College, for their kind help and support for doing such practical work.

 

CONFLICT OF INTEREST:

The author declares no conflict of interest.

 

Abbreviation:

ADQ – Amodiquine Hydrochloride

QbD – Quality by Design

AQbD – Analytical Quality by Design

LOD – Limit of detection

LOQ –Limit of Quantification

 

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Received on 14.03.2019                 Accepted on 21.04.2019

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Asian J. Pharm. Ana. 2019; 9(3):113-117.