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Extraction of proposed IAA biosynthesis intermediates: Anthranilate, Ser, IAAld, IPyA, IAOx, IAN, IAM, indole

MT Molly Tillmann
QT Qian Tang
JC Jerry D. Cohen
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Homogenized samples are incubated on ice for 50 minutes to allow isotopic standard equilibration with the endogenous compounds, diluted 10-fold with water to allow proper interaction with the solid phase, centrifuged to remove solids, and loaded onto a SPE micro spin column (TopTips) containing hydrophilic-lipophilic balanced (HLB) resin conditioned with acetonitrile followed by 20% acetonitrile in water. After loading, the spin columns are washed with 5% acetonitrile and compounds are eluted with 80% acetonitrile.

RENSA HLB resin (MIP Technologies, 92,001–0010) suspended in methanol, 1:5 w:v

Acetonitrile: 100%, 80%, 20%, and 5% prepared in distilled water

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