A thin-layer chromatography-bioautographic method for detecting dipeptidyl peptidase IV inhibitors in plants

A thin-layer chromatography (TLC)-bioautographic method was developed with the aim to detect dipeptidyl peptidase IV (DPP IV) inhibitors from plant extracts. The basic principle of the method is that the enzyme (DPP IV) hydrolyzes substrate (Gly-Pro-p-nitroaniline) into p-nitroaniline (pNA), which diazotizes with sodium nitrite, and then reacts with N-(1-naphthyl) ethylenediamine dihydrochloride in turn to form a rose-red azo dye which provides a rose-red background on the TLC plates. The DPP IV inhibitors showed white spots on the background as they blocked enzymolysis of the substrate to produce pNA. The method was validated with respect to selectivity, sensitivity, linearity, precision, recovery, and stability after optimizing key parameters including plate type, time and temperature of incubation, concentration of substrate, enzyme and derivatization reagents, and absorption wavelength. The results showed good lineary within amounts over 0.01–0.1 μg range for the positive control, diprotin A, with the coefficient of determination (r2) = 0.9668. The limits of detection (LOD) and quantification (LOQ) were 5 and 10 ng, respectively. The recoveries ranged from 98.9% to 107.5%. The averages of the intra- and inter-plate reproducibility were in the range of 4.1–9.7% and 7.6–14.7%, respectively. Among the nine methanolic extracts of medicinal herbs screened for DPP IV inhibitors by the newly developed method, Peganum nigellastrum Bunge was found to have one white active spot, which was then isolated and identified as harmine. By spectrophotometric method, harmine hydrochloride was found to have DPP-IV inhibitory activity of 32.4% at 10 mM comparing to that of 54.8% at 50 μM for diprotin A.

Collectors, Producers, and Circulators of Tibetan and Chinese Medicines in Sichuan Province

The act of prescribing pharmaceutical drugs to patients is normally the site of judgements about the drug’s efficacy and safety. The success of treatments and the licences for commodities depend on the biochemical identity of the drugs and of their path and transformations inside the body. However, the ‘supply chain’ outside the body is eschewed by such discourse, and its importance for both pharmaceutical brands and physician-centred historiographies is ignored. As this ethnographic fieldwork on Tibetan and Chinese medicines in Sichuan shows, overlooked social actors ensure reliable knowledge about medicinal things and materials long before patients take their medicine. This paper takes a step back from the final products—clearly defined as ‘Tibetan’ or ‘Chinese’—and introduces those who produce and distribute them. Via observations of particular regimes of circulation and processing, the actions of collecting, manufacturing, transporting, and educating appear as the first and foremost acts of efficacy and safety.