Poly(acrylic acid) Hydrogel Microspheres for a Metal-Ion Sensor

Abstract

Monodispersed cross-linked poly(acrylic acid) (PAA) droplets (PAA X-droplets), prepared using the microfluidic method with in situ ultraviolet curing, were used as small spherical sensors to simultaneously detect both Ca²⁺ and Mg²⁺ in human saliva and serum. The PAA X-droplet treated with KOH (PAA_KOH X-droplet) was used as a reference droplet because of its highly swollen state. The PAA_KOH X-droplets shrunk in response to the presence of divalent metal ions (Ms) by forming a bridged structure of COO-M-OOC. The sizes of the PAA_KOH X-droplets were precisely and dynamically monitored in the poly(dimethylsiloxane) (PDMS) channel with passing time when the aqueous metal-ion solutions were flowing at a controlled flow rate. The sizes of the PAA_KOH X-droplets continuously decreased to the saturated constant size. The saturated size of the PAA_KOH X-droplet did not change; however, the speed of size reduction increased with an increase in the concentration of the divalent metal ion. The saturated size was studied using the saturated diameter ratio (R_sat-dia) with respect to the initial diameter of the PAA_KOH X-droplet before the metal-ion treatment, and the speed of the size reduction was investigated using the inverse time to reach half the saturated diameter reduction (T_1/2^-1). Ca²⁺ and Mg²⁺ exhibited R_sat-dia values of 75.9 and 83.6%, respectively, when the flow rate was 5 μL min^-1, regardless of the metal concentration. The T_1/2^-1s for the Ca²⁺ and Mg²⁺ linearly increased with an increase in their concentrations. The R_sat-dia of the aqueous Ca²⁺/Mg²⁺ mixture solution had a linear relationship with φ [= C_Ca/(C_Ca + C_Mg), where C_Ca and C_Mg are the molar concentrations of Ca²⁺ and Mg²⁺, respectively]. The T_1/2^-1 of the aqueous Ca²⁺, Mg²⁺ mixture solution was calculated by adding the individual T_1/2^-1s of pure aqueous Ca²⁺ and Mg²⁺ solutions. Using the R_sat-dia and T_1/2^-1 of the Ca²⁺/Mg²⁺ mixture aqueous solution, the individual C_Ca and C_Mg in the mixture solution were successfully calculated. This method was applied to the human saliva and serum in which the major metal ions are Ca²⁺ and Mg²⁺, and other metal ions existed in undetectable amounts by the PAA_KOH X-droplets. This method is simple, cost-effective, and highly accurate and solves the hurdles of separating the interference effect of a Mg²⁺ ion when a Ca²⁺ ion is measured in biofluids.

Keywords: biosensor; calcium; hydrogel; magnesium; microfluidics; poly(acrylic acid).