One-shot screen-printed thylakoid membrane-based biosensor for the detection of photosynthetic inhibitors in discrete samples

Screen-printing technology offers the possibility to produce a large number of sensors at low cost. Thus, due to their intrinsic characteristics and reproducibility, screen-printed electrodes can be used in the development of disposable electrochemical devices. In the present work, carbon-based screen-printed electrodes (SPCEs) have been used to develop a one-shot-measure biosensor for the detection of photosynthetic inhibitors in discrete samples. The measurement was based on the electrochemical evaluation of the activity of photosystem II (PSII), a protein complex present in photosynthetic organisms and involved in the photosynthesis. The biosensor was prepared by the modification of the working electrode of a SPCE, using thylakoid membranes extracted from spinach leaves. The modified sensors were then used as one-shot system to measure the presence of PSII activity inhibitors in discrete standard solutions. The coupling of the developed biosensor with a custom-made cell made it possible to perform tests using only 50 (mu)L of total sample volume with a measurement time of 10 min. Inhibition curves were recorded for some photosynthetic inhibitors in a concentration range of 10^-6 to 10^-8 mol L^-1. A reproducibility (relative standard deviation, R.S.D.%) of 10% was found and the calculated limit of detections (LODs) were in the nanomolar range. The effect of storage on sensitivity and reproducibility of a biosensor prepared by direct lyophilisation of thylakoid membranes on the electrode surface was also evaluated, confirming the possible use of the modified sensor up to one week after the preparation. Measurements on real samples were also reported, comparing the results with those obtained using a fluorescence-based commercial instrument for the analysis of photosynthetic inhibitors.