HPLC Method for Analysis of Red Dye from Marked Pectinophora gossypiella (Lepidoptera: Gelechiidae)

Nada Tomic-Carruthers, Noreen Mahoney, Michelle Walters, John Claus, Guolei Tang

Abstract


We describe an HPLC analytical technique for detection of minute amounts of oil-based Red 26 dye used for marking insects. Chromatographic separation was achieved by an isocratic elution on an Inertsil 100A, 5μ silica column using hexane: isopropyl alcohol (98:2) as the mobile phase and with absorption detection at 520 nm. This newly developed technique is fast and very sensitive, with a detection threshold at 0.1 ng and quantitation down to 0.3 ng per pink bollworm adult. Using the newly developed technique we were able to correlate the titer of dye extracted from adult moths to the amount of dye included in the larval diet. This method is currently accepted by the U.S. cotton industry as a preferred technique for distinguishing artificial diet-reared sterile from wild pink bollworm moths.

 

Se describe una tecnica de HPLC analitica para la deteccion de pequenas cantidades de tinte Rojo 26 de base de aceite utilizado para marcar los insectos. Se logro la separacion cromatografica mediante una elucion isocratica en una columna Inertsil 100A de silice de 5μ usando hexano: alcohol isopropilico (98:2) como fase movil y con la deteccion de la absorcion a 520 nm. Esta tecnica recientemente desarrollada es rapida y muy sensible, con un umbral de deteccion a 0.1 ng y una cuantificacion hasta 0.3 ng por los adultos del gusano rosado del algodon. Utilizando esta nueva tecnica pudimos correlacionar el titulo de la tinta extraida de las polillas adultas a la cantidad de tinta incluida en la dieta de las larvas. Este metodo es aceptado actualmente por la industria del algodon de EEUU como una tecnica preferida para distinguir las polillas esteriles del gusano rosado alimentadas con una dieta artificial, de las polillas salvajes.

 

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Keywords


sterile insect technique (SIT); pink bollworm; rearing; chromatography; cotton

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