DEVELOPMENT AND VALIDATION OF SPECTROSCOPIC METHOD FOR SIMULTANEOUS ESTIMATION OF PALIPERIDONE AND OLANZAPINE IN SYENTHETIC MIXTURE

A simple, specific, accurate, precise and economical spectrophotometric method has been developed for the estimation of Paliperidone and Olanzapine in synthetic mixture. The absorption maxima of the drugs were found to be 269nm and 259nm for Paliperidone and Olanzapine respectively. Paliperidone and Olanzapine obeyed beer’s law in the concentration range 3-18 μg/ml and 2-12 μg/ml respectively. Different analytical parameter such as linearity, precision, accuracy, limit of detection (LOD), limit of quantitation (LOQ), were determined as per ICH guidelines. Hence, the developed method can be used quality control analysis.

A synthetic mixture of Paliperiodne and Olanzapine is the synergistic effect was observed by improving psychotic symptoms. Literature survey several method have been developed for the determination of Paliperidone and Olanzapine each single formulation. The work is the UV spectroscopic method of Paliperidone and Olanzapine in synthetic mixture.
A synthetic mixture of Paliperiodne and Olanzapine is the synergistic effect was observed by improving psychotic symptoms. Literature survey several method have been developed for the determination of Paliperidone and Olanzapine each single formulation. The work is the UV spectroscopic method of Paliperidone and Olanzapine in synthetic mixture.

Structure
Available Online at http://www.recentscientific.com International Journal of
A synthetic mixture of Paliperiodne and Olanzapine is the synergistic effect was observed by improving psychotic symptoms. Literature survey several method have been developed for the determination of Paliperidone and Olanzapine each single formulation. The work is the UV spectroscopic method of Paliperidone and Olanzapine in synthetic mixture.

Structure
Available Online at http://www.recentscientific.com International Journal of

Instrumentation
A double beam UV-VIS spectrophotometer (UV-1800, Shimadzu) connected to computer loaded with spectra manager software UV probe was employed with spectral band width of 1nm and wavelength accuracy of ± 0.3nm with a pair of 10 mm matched quartz cells. All weights were taken on electronic balance.

Chemicals and reagents
Methanol used as diluent. Drug sample of Paliperidone and Olanzapine was given as a gift sample by Intas pharmaceutical ltd and cadila pharmaceutical ltd. Tablets of Paliperidone and Olanzapine were produced from local market.

Selection of a Solvent
Methanol was selected as solvent for studying spectral characteristic of drug.

Preparation of standard stock solution
Preparation of standard stock solution of Paliperidone (100 µg/ml): Accurately weigh 10 mg of Paliperidone was transferred into a 100 ml volumetric flask and diluted with Methanol. Preparation of standard stock solution of Olanzapine (100 µg/ml): Accurately weigh 10 mg of Olanzapine was transferred into a 100 ml volumetric flask and diluted with Methanol.

Selection of Analytical Wavelength
Standard 3-18 µg/ml solution of Paliperidone and 2-12 µg/ml solution of Olanzapine were prepared in methanol by appropriate dilution and spectrum was recorded between 200-400 nm. All zero order spectrum (D 0 ) were converted to first derivative spectrum (D 1 ) using delta lambda 2.0 and scaling factor 10. The overlain first derivative spectrums of Paliperidone and Olanzapine at different concentration were recorded. The zero crossing point (ZCP) of Paliperidone and Olanzapine found to be 269 nm and 259 nm.

Linearity
Linearity evaluates the analytical procedure's ability (within a given range) to obtain a response that is directly to the concentration (amount) of analyte standard. If the method is linear, the test results are directly, proportional to the concentration of analyte in samples within a given range. A calibration curve is prepared by plotting absorbance (Y) as a function of concentration (X) which procedures a linear curve with correlation equation. Y=m X +c Coefficient of determination (r 2 ) should be 0.9999.

Range
Range is the interval between the upper and lower concentration (amounts) of analyte in the sample (including these concentrations) for which it has been demonstrated that the analytical procedure has a suitable level of precision, accuracy and linearity. Coefficient of determination (r 2 ) should be 0.9998.

Accuracy (recovery)
Accuracy expresses the closeness agreement between the value found and the value that is accepted as either a conventional true value or an accepted reference value. It may often be expressed as the recovery by the assay of known, added amounts of analyte.
Coefficient of determination (r 2 ) should be greater than 0.

Precision
Validation of tests for assay and for quantitative determination of impurities includes an investigation of precision.

Repeatability
Repeatability evaluates the variation by a single analyst on a single instrument. Repeatability does not distinguish between variation from the instrument or system alone and from the sample preparation process. Repeatability is performed by analyzing multiple replicates of an assay composite sample using the analytical method. The recovery value is calculated and reported foe each value.

Intermediate Precision
Intermediate precision refers to variation within a laboratory as with in different days, with different instruments, by different analysts, and so forth intermediate precision was formally known as ruggedness. A statistical comparison is made to the first analyst's results.

Instrumentation
A double beam UV-VIS spectrophotometer (UV-1800, Shimadzu) connected to computer loaded with spectra manager software UV probe was employed with spectral band width of 1nm and wavelength accuracy of ± 0.3nm with a pair of 10 mm matched quartz cells. All weights were taken on electronic balance.

Chemicals and reagents
Methanol used as diluent. Drug sample of Paliperidone and Olanzapine was given as a gift sample by Intas pharmaceutical ltd and cadila pharmaceutical ltd. Tablets of Paliperidone and Olanzapine were produced from local market.

Selection of a Solvent
Methanol was selected as solvent for studying spectral characteristic of drug.

Preparation of standard stock solution
Preparation of standard stock solution of Paliperidone (100 µg/ml): Accurately weigh 10 mg of Paliperidone was transferred into a 100 ml volumetric flask and diluted with Methanol. Preparation of standard stock solution of Olanzapine (100 µg/ml): Accurately weigh 10 mg of Olanzapine was transferred into a 100 ml volumetric flask and diluted with Methanol.

Selection of Analytical Wavelength
Standard 3-18 µg/ml solution of Paliperidone and 2-12 µg/ml solution of Olanzapine were prepared in methanol by appropriate dilution and spectrum was recorded between 200-400 nm. All zero order spectrum (D 0 ) were converted to first derivative spectrum (D 1 ) using delta lambda 2.0 and scaling factor 10. The overlain first derivative spectrums of Paliperidone and Olanzapine at different concentration were recorded. The zero crossing point (ZCP) of Paliperidone and Olanzapine found to be 269 nm and 259 nm.

Linearity
Linearity evaluates the analytical procedure's ability (within a given range) to obtain a response that is directly to the concentration (amount) of analyte standard. If the method is linear, the test results are directly, proportional to the concentration of analyte in samples within a given range. A calibration curve is prepared by plotting absorbance (Y) as a function of concentration (X) which procedures a linear curve with correlation equation. Y=m X +c Coefficient of determination (r 2 ) should be 0.9999.

Range
Range is the interval between the upper and lower concentration (amounts) of analyte in the sample (including these concentrations) for which it has been demonstrated that the analytical procedure has a suitable level of precision, accuracy and linearity. Coefficient of determination (r 2 ) should be 0.9998.

Accuracy (recovery)
Accuracy expresses the closeness agreement between the value found and the value that is accepted as either a conventional true value or an accepted reference value. It may often be expressed as the recovery by the assay of known, added amounts of analyte.

Coefficient of determination (r 2 ) should be greater than 0.9998. Precision
Validation of tests for assay and for quantitative determination of impurities includes an investigation of precision.

Repeatability
Repeatability evaluates the variation by a single analyst on a single instrument. Repeatability does not distinguish between variation from the instrument or system alone and from the sample preparation process. Repeatability is performed by analyzing multiple replicates of an assay composite sample using the analytical method. The recovery value is calculated and reported foe each value.

Intermediate Precision
Intermediate precision refers to variation within a laboratory as with in different days, with different instruments, by different analysts, and so forth intermediate precision was formally known as ruggedness. A statistical comparison is made to the first analyst's results.

Instrumentation
A double beam UV-VIS spectrophotometer (UV-1800, Shimadzu) connected to computer loaded with spectra manager software UV probe was employed with spectral band width of 1nm and wavelength accuracy of ± 0.3nm with a pair of 10 mm matched quartz cells. All weights were taken on electronic balance.

Chemicals and reagents
Methanol used as diluent. Drug sample of Paliperidone and Olanzapine was given as a gift sample by Intas pharmaceutical ltd and cadila pharmaceutical ltd. Tablets of Paliperidone and Olanzapine were produced from local market.

Selection of a Solvent
Methanol was selected as solvent for studying spectral characteristic of drug.

Preparation of standard stock solution
Preparation of standard stock solution of Paliperidone (100 µg/ml): Accurately weigh 10 mg of Paliperidone was transferred into a 100 ml volumetric flask and diluted with Methanol. Preparation of standard stock solution of Olanzapine (100 µg/ml): Accurately weigh 10 mg of Olanzapine was transferred into a 100 ml volumetric flask and diluted with Methanol.

Selection of Analytical Wavelength
Standard 3-18 µg/ml solution of Paliperidone and 2-12 µg/ml solution of Olanzapine were prepared in methanol by appropriate dilution and spectrum was recorded between 200-400 nm. All zero order spectrum (D 0 ) were converted to first derivative spectrum (D 1 ) using delta lambda 2.0 and scaling factor 10. The overlain first derivative spectrums of Paliperidone and Olanzapine at different concentration were recorded. The zero crossing point (ZCP) of Paliperidone and Olanzapine found to be 269 nm and 259 nm.

Linearity
Linearity evaluates the analytical procedure's ability (within a given range) to obtain a response that is directly to the concentration (amount) of analyte standard. If the method is linear, the test results are directly, proportional to the concentration of analyte in samples within a given range. A calibration curve is prepared by plotting absorbance (Y) as a function of concentration (X) which procedures a linear curve with correlation equation. Y=m X +c Coefficient of determination (r 2 ) should be 0.9999.

Range
Range is the interval between the upper and lower concentration (amounts) of analyte in the sample (including these concentrations) for which it has been demonstrated that the analytical procedure has a suitable level of precision, accuracy and linearity. Coefficient of determination (r 2 ) should be 0.9998.

Accuracy (recovery)
Accuracy expresses the closeness agreement between the value found and the value that is accepted as either a conventional true value or an accepted reference value. It may often be expressed as the recovery by the assay of known, added amounts of analyte.
Coefficient of determination (r 2 ) should be greater than 0.

Precision
Validation of tests for assay and for quantitative determination of impurities includes an investigation of precision.

Repeatability
Repeatability evaluates the variation by a single analyst on a single instrument. Repeatability does not distinguish between variation from the instrument or system alone and from the sample preparation process. Repeatability is performed by analyzing multiple replicates of an assay composite sample using the analytical method. The recovery value is calculated and reported foe each value.

Intermediate Precision
Intermediate precision refers to variation within a laboratory as with in different days, with different instruments, by different analysts, and so forth intermediate precision was formally known as ruggedness. A statistical comparison is made to the first analyst's results.
Xi = individual measurement in a set X = arithmetic mean of the set and N = number of replicates taken in the set RSD = SD/X % RSD or coefficient or variance (CV) is expressed as 16561 | P a g e %RSD = CV = SD/X*100 The % RSD of the assay/recovery values generated by a single analyst should not be greater than 2.0%. The %RSD of the combined assay/recovery values generated by both analyst, over both analyst should not be greater than 2.0%. The %RSD of the related compounds recovery value should not be greater than 15%.

Detection limit
The detection limit (DL or limit of detection LOD) of an individual procedure is the lowest of analyte in a sample that can be detected but not necessarily quantitated as an exact value.
The detection limit (DL) may be expressed as: DL = 3.3*σ/S Where, S= the slope of calibration curve σ = Standard deviation of blank reading or intercepts of calibration curve

Quantitation limit
The quantitation limit (QL or limit of detection LOQ) of an individual procedure is the lowest amount of analyte in a sample that can be quantitatively determined with suitable precision and accuracy. The quantitation limit (QL) may be expressed as: QL = 10*σ/s Where, S= the slope of calibration curve σ = Standard deviation of blank reading or intercepts of calibration curve

RESULT AND DISCUSSION
A reliable first order derivative spectrophotometric method was developed for simultaneous estimation of Paliperidone and Olanzapine in syenthetic mixture by UV spectrophotometry. Beer's law was obeyed in concentration range of 3-18 µg/ml for Paliperidone and 2-12 µg/ml for Olanzapine at 269 nm and 259 nm wavelengths, respectively. The correlation coefficient Paliperidone and Olanzapine was found to be R 2 = 0.998 and 0.999. The mean % recoveries were found to be in the range of 99.2%-101.99% and 99%-102%, respectively. The LOD and LOQ were 0.2131 µg/ml and 0.645 µg/ml of Paliperidone and 0.218 µg/ml and 0.662 µg/ml of Olanzapine, respectively. The proposed method was precise, accurate and reproducible and acceptable recovery of the analyses, which can be applied for the analysis of Paliperidone and Olanzapine in syenthetic mixture.

CONCLUSION
A simple accurate and precise UV spectrophotometric method has been developed for the estimation of Paliperidone and Olanzapine in synthetic mixture. It has advantage that it eliminates the spectral interference from one of the two drugs while estimating the other drug at selected wavelength.