Determination of 1-deoxynojirimycin in Morus alba L. leaves by derivatization with 9-fluorenylmethyl chloroformate followed by reversed-phase high-performance liquid chromatography

• Published in: Journal of Chromatography A 2003-06, Vol.1002 (1), p.93-99
• Main Authors: Kim, Jin-Won, Kim, Soo-Un, Lee, Heui Sam, Kim, Iksoo, Ahn, Mi Young, Ryu, Kang Sun
• Format: Article
• Language: English
• Subjects: 1-Deoxynojirimycin - analysis; Alkaloids; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Deoxynojirimycin; Fluorenes - chemistry; General pharmacology; Medical sciences; Morus - chemistry; Pharmacognosy. Homeopathy. Health food; Pharmacology. Drug treatments; Plant Leaves - chemistry; Sensitivity and Specificity
• ISSN: 0021-9673
• DOI: 10.1016/S0021-9673(03)00728-3

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 °C). Linearity ranged between 0.3 and 30 μg 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.