A prospective method for non-invasive diagnostic of iron deficiency anemia using microfluidic paper-based analytical devices with a natural reagent from red rose (Rosa damascena) flower extract

Abstract

A non-invasive, low-cost, non-toxic, and user-friendly microfluidic paper-based analytical device (µPADs) was developed for Fe(II) determination in artificial urine samples. The determination method is based on the color change of anthocyanin from Rose damascena flower extract (RDFE) through the formation of a Fe(II)-anthocyanin complex. The image was captured using a smartphone camera and analyzed for color intensity using ImageJ. The "Mickey Mouse" hydrophobic pattern on µPADs was created by printing a pattern consisting of one circular sample zone, two circular detection zones of the same size (7.5 mm i.d.), and two-channel zones with dimensions of 3.3 x 2.5 mm. The optimal conditions for µPADs were 1.2 µL of extract (8% v/v), 28 µL of sample, and a reaction time of 25 minutes. The proposed µPADs method offers a linear dynamic range of 0 to 4 mg/L, with LOD and LOQ values of 0.075 and 0.25 mg/L, respectively. The validation of the method using recovery tests yields results ranging from 93% to 106%, which is considered accurate. This method successfully determined Fe(II) ions in artificial urine samples with Fe(II) levels mimicking those of patients with iron deficiency anemia, and it is highly applicable to other sample types.