DETERMINATION OF SN-38 CONTENTS IN NANOCAPSULES BY CHROMATOGRAPHIC AND SPECTROPHOTOMETRIC METHODS
Shilpi Prasad* and J. S. Dangi
ABSTRACT
A rapid and sensitive reversed phase high performance liquid
chromatography (RP-HPLC) method using isocratic elution with UV
detection and UV spectrophotometric method was developed for
quantification of SN-38 (7-ethyl-10-hydroxycamptothecin), a
camptothecin derivative in a poly (lactic-co-glycolic acid) (PLGA)
based nanocapsule (PLNP). Chromatographic separation was
performed on a Kromasil,KR100,C18, (4.6 mm X 250 mm, 5μm)
analytical column with a mobile phase containing a mixture of octane-
1-sulfonic acid sodium salt, potassium di hydrogen phosphate and
acetonitrile (29:29:42v/v/v). SN-38 was detected at a wavelength of
265nm. The limit of detection (LOD) and limit of quantitation (LOQ)
of HPLC method were found to be 0.006 μg mL-1 and 0.02 μg mL-1 for
carboxylate and 0.002 μg mL-1 and 0.02 μg mL-1 for lactone form respectively. For UV
method LOD and LOQ is 0.026 μg mL-1 and 0.08 μg mL-1 respectively. The methods were
validated for linearity, accuracy, precision and robustness as per International Conference on
Harmonization (ICH) guidelines and applied for determination of entrapment efficiency and
drug loading in nanocapsules during in vitro analysis. Although HPLC method is more
specific and precise and has better detection and quantification limits than UV method but
quantitative determination by UV method becomes important when solvent recovery, cost
effectiveness and time of analysis is important.
Keywords: HPLC, UV spectroscopy, Nanocapsules, 7-ethyl-10-hydroxycamptothecin, poly (lactic-co-glycolic acid).
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