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Abstract
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Microfluidic separation technologies have emerged as powerful alternatives to traditional chromatography, addressing issues such as high costs, lengthy
processing times, and low efficiency [1, 2]. Cyclotides, which are plant-derived cyclic peptides with a wide range of biological activity from antibacterial
to anticancer, require efficient purifying procedures before they can be used in drug development. We provide the first example of a microfluidic device
that replicates high-performance liquid chromatography (HPLC) for cyclotide separation. Silica gel-C18 was produced, analyzed, and incorporated into
microchannels to form an HPLC-like stationary phase. Cyclotide extracts from Viola ignobilis were prepared using low-voltage electric field extraction,
evaluated using HPLC and MALDI-TOF, and then put into the microfluidic system. Using distilled water as the mobile phase, several cyclotides (Vigno 1-
5 and Varv A) were successfully separated and collected at varying retention times. These findings demonstrate the promise of silica gel-C18 microfluidic
chips as a fast, low-cost, and effective technology for cyclotide separation, with significant implications for accelerating natural product research and drug
discovery [3, 4, 5].
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