On – line immunoaffinity capture, coupled with HPLC and electrospray innization mass spectrometry, for automated determination of fumonisins.
NEWKIRK, D.K., BENSON, R.W., HOWARD, P.C., CHURCHWELL, M.I., DOERGE, D.R. and ROBERTS, D.W.
Journal of Agricultural and Food Chemistry 46: 1677 – 1688.
1998
บทคัดย่อ
An automated online method for the quantitative detection of AFB1, B2 and B3 and hydrolysed FB1 in corn based feed was developed using a combination of immunoaffinity capture/ cleanup, coupled with reverse phase LC and electrospray ionisation – MS. Bocking the C2 amine of FB1 with 9 – fluorenylmethylchloroformate allowed coupling to keyhole limpet haemocyanin through the tricarballylic end, leaving the amino terminus exposed on the immunogenic conjugate after removal of the blocking group. Antiserum had specificity for the C1 – C10 fumonisin domain and was used for ELISA and to prepare immunoaffinity columns. The quantitation limit was 250 pg using the protonated molecule signal. Similar sensitivity was observed for FB2 and B3 and hydrolysd FB1. The online IAC/LC/ESI –MS method provided a linear response from the detection limit to 5 ng.