Determination of 1-deoxynojirimycin in Morus alba L. leaves by derivatization with 9-fluorenylmethyl chloroformate followed by reversed-phase high-performance liquid chromatography.
A rapid and reliable method suitable for assays of a large number of Morus alba leaves for 1-deoxynojirimycin (DNJ) has been developed. DNJ in 0.1 g of freeze-dried leaves was double-extracted in 10 mL of aqueous 0.05 M HCl by vortexing for 15 s at room temperature, derivatized with 9-fluorenylmethyl chloroformate (FMOC-Cl), and analyzed by reversed-phase high-performance liquid chromatography (RP-HPLC) equipped with a fluorescence detector. The double extraction recovered > 99% of extractable DNJ from the leaves. Stabilization of FMOC-derivatized DNJ (DNJ-FMOC) was achieved by diluting the reactant with aqueous acetic acid after derivatization. DNJ-FMOC was stable for at least 16 days under acidic conditions at room temperature (24 degrees C). Linearity ranged between 0.3 and 30 microg mL(-1). The intra- and inter-day precision for DNJ-spiked biological samples was between 0.6 and 1.8% and between 3.7 and 4.5%, respectively.
Kim
Jin-Won
JW
Department of Sericulture and Entomology, National Institute of Agricultural Science and Technology, Suwon 441-100 South Korea. annua38@empal.com
Kim
Soo-Un
SU
Lee
Heui Sam
HS
Kim
Iksoo
I
Ahn
Mi Young
MY
Ryu
Kang Sun
KS
D017485:1-Deoxynojirimycin / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005449:Fluorenes / Q000737:chemistry*; D031625:Morus / Q000737:chemistry*; D018515:Plant Leaves / Q000737:chemistry*; D012680:Sensitivity and Specificity
D005449:Fluorenes; D017485:1-Deoxynojirimycin; C054007:1-(9-fluorenyl)methyl chloroformate