Rapid in-syringe-based ultrasonic-energy assisted salt-enhanced homogeneous liquid-liquid microextraction technique coupled with HPLC/ low-temperature evaporative light-scattering detector for quantification of sodium hyaluronate in food products

Sodium-hyaluronate (Na-HA) is a biopolymer-based active ingredient in various dietary and beauty care supplements. However, the existing analytical methods for Na-HA analysis are laborious, less sensitive, and timeconsuming. This study demonstrates a simple, low-cost, and green rapid analytical methodology using a novel in-syringe-based ultrasonic energy-assisted salt-enhanced homogeneous liquid-liquid microextraction (IS-USESE-HLLME) technique coupled with HPLC/low-temperature evaporative light scattering detector (LT-ELSD) for the sensitive quantification of Na-HA in nutraceutical products. This method utilizes ultrasound energy to extract Na-HA from an aqueous sample solution using a low volume water-miscible extraction solvent with a high salt concentration. Size-exclusion column-based HPLC was applied to separate target Na-HA from co-matrices and quantified using LT-ELSD for sensitive analysis. Under optimal conditions, excellent linearity was observed between 1 and 500 ng mL-1 with a correlation coefficient of 0.9974. Detection and quantification limits were 0.5 ng mL-1 and 1.0 ng mL-1, respectively. The developed (IS-USA-SE-HLLME/HPLC-LT-ESLD) method was applied to analyze Na-HA in commercial nutraceutical and beauty care products. Spiked recoveries of real samples showed between 96.1 and 105.9%, with the precision <7.52%, indicating no matrices influence and good accuracy of the method. These results proved that the developed method is a fast, inexpensive, sensitive, and ecofriendly method for quantifying Na-HA in nutraceutical products. Therefore, the presented method can be applied as an alternative standard analytical method to monitor Na-HA concentrations in food products for food safety regulatory purposes.

Automated summary

The study introduced a rapid analytical method for the sensitive quantification of Na-HA in nutraceutical products, utilizing an innovative ultrasonic energy-assisted salt-enhanced liquid-liquid microextraction technique coupled with HPLC/LT-ELSD. The method showed excellent linearity, low detection and quantification limits, and good accuracy and precision when applied to analyze commercial products. This method offers a fast, inexpensive, sensitive, and ecofriendly approach to monitoring Na-HA concentrations in food products.