Box–Behnken design aided optimization and validation of developed reverse phase HPLC analytical method for simultaneous quantification of dolutegravir sodium and lamivudine co‐loaded in nano‐liposomes
A stability‐indicating reversed‐phase high‐performance liquid chromatography method for simultaneous estimation of dolutegravir sodium and lamivudine encapsulated in the nanoliposomal formulation was developed. The chromatographic parameters namely, organic phase ratio, flow rate, and sample injecti...
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| Veröffentlicht in: | Journal of separation science Jg. 44; H. 15; S. 2917 - 2931 |
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| Abstract | A stability‐indicating reversed‐phase high‐performance liquid chromatography method for simultaneous estimation of dolutegravir sodium and lamivudine encapsulated in the nanoliposomal formulation was developed. The chromatographic parameters namely, organic phase ratio, flow rate, and sample injection volume were selected as independent factors and were optimized by multivariate Box–Behnken design. Responses analyzed were retention time, peak area, and resolution. The optimized chromatographic method with Hypersil BDS C8 CN column as stationary phase and methanol and acetonitrile mixture and acidified Milli‐Q water (pH 2.8, adjusted with 0.02% v/v orthophosphoric acid) as the mobile phase in an isocratic elution mode was validated according to parameters of International Conference on Harmonization Q1(R2) guidelines. The validated reversed‐phase high‐performance liquid chromatography method exhibited specificity for both dolutegravir sodium and lamivudine in the presence of degradation products as well as the liposomal matrix. This method was effectively utilized to determine the amount of drug entrapped and drug loading efficiency of dolutegravir sodium and lamivudine in a nano‐liposomal formulation. |
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| AbstractList | A stability-indicating reversed-phase high-performance liquid chromatography method for simultaneous estimation of dolutegravir sodium and lamivudine encapsulated in the nanoliposomal formulation was developed. The chromatographic parameters namely, organic phase ratio, flow rate, and sample injection volume were selected as independent factors and were optimized by multivariate Box-Behnken design. Responses analyzed were retention time, peak area, and resolution. The optimized chromatographic method with Hypersil BDS C8 CN column as stationary phase and methanol and acetonitrile mixture and acidified Milli-Q water (pH 2.8, adjusted with 0.02% v/v orthophosphoric acid) as the mobile phase in an isocratic elution mode was validated according to parameters of International Conference on Harmonization Q1(R2) guidelines. The validated reversed-phase high-performance liquid chromatography method exhibited specificity for both dolutegravir sodium and lamivudine in the presence of degradation products as well as the liposomal matrix. This method was effectively utilized to determine the amount of drug entrapped and drug loading efficiency of dolutegravir sodium and lamivudine in a nano-liposomal formulation.A stability-indicating reversed-phase high-performance liquid chromatography method for simultaneous estimation of dolutegravir sodium and lamivudine encapsulated in the nanoliposomal formulation was developed. The chromatographic parameters namely, organic phase ratio, flow rate, and sample injection volume were selected as independent factors and were optimized by multivariate Box-Behnken design. Responses analyzed were retention time, peak area, and resolution. The optimized chromatographic method with Hypersil BDS C8 CN column as stationary phase and methanol and acetonitrile mixture and acidified Milli-Q water (pH 2.8, adjusted with 0.02% v/v orthophosphoric acid) as the mobile phase in an isocratic elution mode was validated according to parameters of International Conference on Harmonization Q1(R2) guidelines. The validated reversed-phase high-performance liquid chromatography method exhibited specificity for both dolutegravir sodium and lamivudine in the presence of degradation products as well as the liposomal matrix. This method was effectively utilized to determine the amount of drug entrapped and drug loading efficiency of dolutegravir sodium and lamivudine in a nano-liposomal formulation. A stability‐indicating reversed‐phase high‐performance liquid chromatography method for simultaneous estimation of dolutegravir sodium and lamivudine encapsulated in the nanoliposomal formulation was developed. The chromatographic parameters namely, organic phase ratio, flow rate, and sample injection volume were selected as independent factors and were optimized by multivariate Box–Behnken design. Responses analyzed were retention time, peak area, and resolution. The optimized chromatographic method with Hypersil BDS C8 CN column as stationary phase and methanol and acetonitrile mixture and acidified Milli‐Q water (pH 2.8, adjusted with 0.02% v/v orthophosphoric acid) as the mobile phase in an isocratic elution mode was validated according to parameters of International Conference on Harmonization Q1(R2) guidelines. The validated reversed‐phase high‐performance liquid chromatography method exhibited specificity for both dolutegravir sodium and lamivudine in the presence of degradation products as well as the liposomal matrix. This method was effectively utilized to determine the amount of drug entrapped and drug loading efficiency of dolutegravir sodium and lamivudine in a nano‐liposomal formulation. |
| Author | Mutalik, Srinivas Mutalik, Sadhana P Mullick, Prashansha Pandey, Abhijeet Kulkarni, Smita S. |
| Author_xml | – sequence: 1 givenname: Sadhana P surname: Mutalik fullname: Mutalik, Sadhana P organization: Manipal Academy of Higher Education – sequence: 2 givenname: Prashansha surname: Mullick fullname: Mullick, Prashansha organization: Manipal Academy of Higher Education – sequence: 3 givenname: Abhijeet surname: Pandey fullname: Pandey, Abhijeet organization: Manipal Academy of Higher Education – sequence: 4 givenname: Smita S. surname: Kulkarni fullname: Kulkarni, Smita S. organization: Division of Virology, ICMR‐National AIDS Research Institute (NARI) – sequence: 5 givenname: Srinivas orcidid: 0000-0002-0642-1928 surname: Mutalik fullname: Mutalik, Srinivas email: ss.mutalik@manipal.edu organization: Manipal Academy of Higher Education |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/34076952$$D View this record in MEDLINE/PubMed |
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| Keywords | liquid chromatography nano-liposomes dolutegravir Box-Behnken design lamivudine |
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| SubjectTerms | Acetonitrile Acidification Antiretroviral drugs Box–Behnken design Chromatography Chromatography, High Pressure Liquid - methods Chromatography, Reverse-Phase - methods Design optimization dolutegravir Drug Carriers Drug Compounding drugs experimental design Flow velocity Heterocyclic Compounds, 3-Ring - analysis High-performance liquid chromatography HIV Integrase Inhibitors - analysis Ions lamivudine Lamivudine - analysis Limit of Detection Liposomes liquid chromatography Mathematical analysis methanol Nanoparticles nano‐liposomes Oxazines - analysis Parameters Phase ratio phosphoric acid Piperazines - analysis Pyridones - analysis Reverse Transcriptase Inhibitors - analysis reversed-phase high performance liquid chromatography separation Sodium |
| Title | Box–Behnken design aided optimization and validation of developed reverse phase HPLC analytical method for simultaneous quantification of dolutegravir sodium and lamivudine co‐loaded in nano‐liposomes |
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