STATISTICAL CORRELATION AND SIMULTANEOUS ESTIMATION OF ATAZANAVIR SULFATE AND RITONAVIR IN FIXED DOSAGE FORM BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY AND HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHY

Abstract

Two chromatographic methods are developed and validated to estimate Atazanavir Sulfate and Ritonavir in new fixed dosage form, that is, tablet dosage form named Synthivan. The first method is based on HPLC separation of the two drugs on the reversed phase HiQSil C18 column (5 µm, 250 × 4.6 mm) at ambient temperature using an isocratic solvent system consisting of acetonitrile and water in the ratio 52:48 (v/v). Quantification is achieved with a PDA detector at 254 nm at a flow rate of 1.5 mL/min. Linearity of concentrations is found at 60–600 µg/mL and 20–200 µg/mL for Atazanavir sulfate and ritonavir, respectively. The second method is based on HPTLC separation of the two drugs on pre-coated silica gel 60F₂₅₄ aluminum plates using toluene:methanol:glacial acetic acid:ethyl acetate (7:0.5:1.5:2, v/v/v/v) as solvent system followed by densitometric measurements of their spots at 254 nm. The linearity of the concentration is found to be 30–300 and 10–100 ng/spot for Atazanavir Sulfate and Ritonavir, respectively. Both methods are validated according to ICH guidelines. The analysis of variance (ANOVA) and Student's t-test are applied to correlate the results of Atazanavir Sulfate and Ritonavir determination in dosage form by means of HPLC and HPTLC method.

Keywords:

• ANOVA
• atazanavir sulfate
• HPLC
• HPTLC
• method development
• ritonavir
• student's t-test
• validation

ACKNOWLEDGMENT

The authors are thankful to Cipla Ltd and Matrix Labs for providing gift samples of Atazanavir sulfate and Ritonavir for the research work. Authors are also thankful to Siksha O Anusandhan University and Bharati Vidyapeeth Deemed University for providing the instrumental facilities for completion of the research work.

Notes

*Mean of six determinations.

*Mean of six determinations.

*R_t, K, and T are retention time, capacity factor, and tailing factor, respectively.

*mean of three determinations.

**mean of six determinations.

^a The results are presented as mg of label claimed amount of ATV in tablets.

^b The results are presented as mg of label claimed amount of RTV in tablets.

F stat < F crit .

^a The results are presented as mg of label claim of ATV in tablet.

^b The results are presented as mg of label claim of RTV in tablet.