Base catalyzed cleavage of 9-fluorenylmethoxycarbonyl (FMOC) group and subsequent analysis by UV spectrophotometry is a commonly used technique for measuring the loading of functional groups on solid supports. Recent works suggest that using 1,8-diazabicyclo[5,4,0]undec-7-ene (DBU) instead of piperidine makes it possible to use gas chromatography for quantitation, but due to the long deprotection time used, the method is not high-throughput. We observed that the dibenzofulvene released after DBU deprotection could be measured by reversed-phase (RP) HPLC. We have optimized the concentration of DBU as well as the time of the deprotection and coupled with a fast RP-HPLC separation results in a highly reproducible, high-throughput method. The measured loading correspond well with the manufacturer's data on several commercial resins. Using this method we have quantitated the amine loading on several polystyrene resins and we have found that the total amount of functional group can be more than twice the amount of the available ones. We concluded that the differences were the function of the resin loading as well as the level of crosslinking.
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