Method development for the determination of biogenic amines in insect hemolymph by HPLC with electrochemical detection.

• DOI: 10.55632/pwvas.v88i1.107
• Publication Date: 2022-05-21T00:58:26Z
• Authors: Nicholas Tucker; Victoria Broadwater; Dan DiLella; Ruth Conley
• Keywords: Solid phase extraction

An HPLC method was developed for the quantification of biogenic amines in insect hemolymph. The major challenge analysis is that a typical sample only contains few hundred pictograms amine. A C18 column and electrochemical detection used. eluent an aqueous 0.06 M phosphate buffer (pH- 6.7) mixed with 10-15% methanol ion-pairing reagent. With this method, can be detected at picogram levels. However, there are hundreds compounds biological fluid and, some cases, another electro-active species co-elute interest. To address problem we have methods to selectively remove potentially interfering by pretreating solid phase extraction (SPE). Previously, had used weak cation SPE columns simplify mixture but found too time consuming. We now uses anion columns. samples were collected syringe preloaded anticoagulant, either trichloroacetic acid or formic acid. proteins removed centrifuging 10K Da filter. followed. This pretreatment effective selective removal many peaks. DHBA (3,4-Dihydroxybenzylamine hydrobromide) as internal standard.