A 3-phase liquid-liquid extraction (LLE) chip was hyphenated with nano-HPLC/ESI-MS (Fig.). Thus the purification, separation and MS detection of alkaloids was accomplished on-line. The system is solvent-efficient with a nanoLC flow rate of 200 nL/min, and chip flow rate of 500 nL/min, sample-efficient (50-330 nL sample volume) and time-efficient (saving tedious off-line sample pretreatment steps. The transfer efficiency of strychnine was above 92% in 25sec). The analyses of alkaloids in Semen strychni, Cephaelis ipecacuanha, Atropa belladonna, Vinca minor, and Coptidis rhizome were successfully performed. The limits of detection (LODs) of the alkaloids are lower than with conventional HPLC/ESI-MS and nano-HPLC-DAD, i.e. 5ng/mL for emetine, palmatine, vincamine, strychnine and 10ng/mL for atropine.