Abstract
A rapid and simple high performance thin layer chromatography (HPTLC) method with densitometry at λ=263 nm was developed and validated for simultaneous determination of lopinavir and ritonavir from pharmaceutical preparation. Separation was performed on aluminum-backed silica gel 60F254HPTLC plates as stationary phase and using a mobile phase comprising of toluene, ethyl acetate, methanol and glacial acetic acid, in the volume ratio of 7.0:2.0:0.5:0.5 (v/v) respectively. After development, plates were observed under UV light. The detector response was linear in the range of 6.67 to 20.00 µg/spot and 1.67 to 5.00 µg/spot for lopinavir and ritonavir respectively. The validated lowest limit of detection was 21.00 ng/spot and 5.10 ng/spot whereas lowest limit of quantification was 7.00 ng/spot and 21.00 ng/spot for lopinavir and ritonavir respectively. The percentage assay of lopinavir and ritonavir was found between 98.23 to 102.28% and 98.03 to 103.50% respectively. The described method has the advantage of being rapid and easy. Hence it can be applied for routine quality control analysis of lopinavir and ritonavir from pharmaceutical preparation and stability studies.
Highlights
Lopinavir1-2 has the molecular formula C37H48N4O5, and molecular weight 628.80 g mol-1
No references are reported so far for the simultaneous determination of both drugs in combined dosage form or any such pharmaceutical preparations by high performance thin layer chromatography (HPTLC). In this communication we report a new simple, rapid and precise HPTLC method for simultaneous determination of lopinavir and ritonavir in combination capsule, which can be used for its routine analysis in ordinary laboratories
The formulation, sustained release capsule with combination of lopinavir 133.3 mg and ritonavir 33.3 mg is available in market by brand name KALETRA; toluene, ethyl acetate, methanol and glacial acetic acid were of Qualigens; pre-coated silica gel 60 F254 HPTLC plates (Merck # 5548) of E-Merck
Summary
Lopinavir has the molecular formula C37H48N4O5, and molecular weight 628.80 g mol-1. Combination therapy with the HIV protease inhibitors lopinavir and ritonavir (Sustained release capsule with combination of lopinavir 133.3 mg and ritonavir 33.3 mg is available in market by brand name kaletra®) has been shown to be effective against drug-resistant HIV-13. The formulation, sustained release capsule with combination of lopinavir 133.3 mg and ritonavir 33.3 mg is available in market by brand name KALETRA; toluene, ethyl acetate, methanol and glacial acetic acid were of Qualigens; pre-coated silica gel 60 F254 HPTLC plates (Merck # 5548) of E-Merck. The drug was dissolved in methanol, diluted up to the mark with methanol and mixed well This gave a standard stock solution of strength 2500 μg/mL of lopinavir. This gave a standard stock solution of strength 1000 μg/mL of ritonavir. The solution was filtered through Whatman No 41 filter paper and the filtrate was used as sample solution
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