Interstitial skin fluid (ISF) is an emerging alternative source of blood samples that has attracted great interest from researchers. It is a very promising way to use microneedle patches for extracting ISF. However, the recovery of ISF still faces great challenges, such as long extraction time and low extraction volume, which may affect the analysis of biomarkers. Traditional centrifugation methods cannot completely recover ISF, which leads to inaccuracy in ISF detection. In this paper, the prepared polyvinyl alcohol/polyvinylpyrrolidone (PVA/PVP) microneedle patches had the ability to insert into the skin in a dry state; at the same time, the microneedle patches had good swelling properties and could extract ISF in a short time without any additional devices. Due to the thermal degradation of PVA, the way of gentle heating was used to recover ISF, which could greatly improve the accuracy of detection. By comparing the D-glucose content assay kit with the blood glucose concentration of rats detected using a commercial glucometer, the detection accuracy of the microneedle patches was verified. The microneedle patches can be used to sample ISF and analyze the level of biomarkers in ISF, and are expected to provide a basis for the prevention and diagnosis of clinical diseases in the future.
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