%0 Journal Article
%T Development and validation of a rapid high performance liquid chromatography - photodiode array detection method for estimation of a bioactive compound wedelolactone in extracts of Eclipta alba
%A Satyanshu Kumar
%A Tushar Dhanani
%J Brazilian Journal of Pharmaceutical Sciences
%D 2013
%I Universidade de S?o Paulo
%X Following optimization of extraction, separation and analytical conditions, a rapid, sensitive and simple reverse-phase high performance liquid chromatography-photo diode array (HPLC-PDA) method has been developed for the identification and quantification of wedelolactone in different extracts of Eclipta alba. The separation of wedelolactone was achieved on a C18 column using the solvent system consisting of a mixture of methanol: water: acetic acid (95: 5: 0.04) as a mobile phase in isocratic elution mode followed by photo diode array detection at 352 nm. The developed method was validated as per the guidelines of the International Conference on Harmonization (ICH). Calibration curve presented good linear regression (r2>0.998) within the test range and the maximum relative standard deviation (RSD, %) values for intra-day assay were found to be 0.15, 1.30 and 1.1 for low (5 ¦Ìg/mL), medium (20 ¦Ìg/mL) and high (80 ¦Ìg/mL) concentrations of wedelolactone. For inter-day assay the maximum RSD (%) values were found to be 2.83, 1.51 and 2.06 for low, medium and high concentrations, respectively. Limit of detection (LOD) and limit of quantification (LOQ) were calculated to be 2 and 5 ¦Ìg/mL respectively. Analytical recovery of wedelolactone was greater than 95%. Wedelolactone in different extracts of Eclipta alba was identified and quantified using the developed HPLC method. The validated HPLC method allowed precise quantitative analysis of wedelolactone in Eclipta. alba extracts. Desenvolveu-se m¨¦todo r¨¢pido, sens¨ªvel e simples de Cromatografia L¨ªquida de Alta Efici¨ºncia em fase reversa, utilizando-se arranjo de fotodiodo (HPLC-PDA), visando ¨¤ separa  o, extra  o e ¨¤s condi  es anal¨ªticas para a identifica  o e quantifica  o de wedelolactona em diferentes extratos de Eclipta alba. A separa  o de wedelolactona foi efetuada por meio de uma coluna C18, utilizando mistura de metanol:¨¢gua:¨¢cido ac¨¦tico (95:5:0.04) como fase m¨®vel, em sistema de elui  o isocr¨¢tica, seguida de detec  o por arranjo de fotodiodo a 352 nm. O m¨¦todo desenvolvido foi validado de acordo com as diretrizes da Confer¨ºncia Internacional de Harmoniza  o (ICH). As curvas de calibra  o apresentaram boa regress o linear (r2>0,998), dentro dos intervalos de teste, e os valores m¨¢ximos de desvio padr o relativo (RSD,%) dos ensaios intra-dia foram 0,15, 1,30 e 1,1 para concentra  es de wedelolactona baixa (5 ¦Ìg/mL), m¨¦dia (20 ¦Ìg/mL) e elevada (80 ¦Ìg/mL) Para o ensaio inter-dia,os m¨¢ximos de RSD (%) foram 2,83, 1,51 e 2,06 para as concentra  es baixa, m¨¦dia e alta, respectivamente. O Limite de Detec  o
%K Cromatografia l¨ªquida de alta efici¨ºncia
%K Cromatografia l¨ªquida de alta efici¨ºncia
%K Cromatografia l¨ªquida de alta efici¨ºncia
%K Eclipta alba
%K Eclipta alba
%K Wedelolactona
%K High performance liquid chromatography
%K High performance liquid chromatography
%K High performance liquid chromatography
%K Eclipta alba
%K Eclipta alba
%K Wedelolactone
%U http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-82502013000100007