Development and validation of a liquid chromatographic method with diode array detection for the determination of...

A HPLC method coupled with diode array detector was developed and validated for the quantitation of alizarin, apigenin, carminic acid, curcumin, ellagic acid, emodin, fisetin, kaempferide, kaempferol, kermesic acid, morin, purpurin, quercetin and sulfuretin which are components of several natural dyes. 1- Hydroxyanthraquinone was selected as internal standard. The compounds were separated under gradient elution on a RP-column (Altima C18, 250 mm x 3.0 mm i.d., 5 μm) with a mobile phase consisting of solvent A: H₂O + 0.1% (v/v) trifluoroacetic acid and solvent B: acetonitrile + 0.1% (v/v) trifluoroacetic acid. The method was validated in terms of linearity, limits of detection and quantitation, accuracy, precision, ruggedness and robustness and applied to the analysis of silk dyed with buckthorn (Rhamnus trees), cochineal (Dactylopius coccus Costa), madder (Rubia tinctorum L.), turmeric (Curcuma longa L.) and young fustic (Cotinus coggygria Scop). Furthermore, dyed silk samples were subjected to artificially accelerated ageing conditions induced by UV radiation. The effect of the latter on the quantities of the aforementioned compounds was monitored, except for apigenin, kermesic acid and morin.