Simple, Rapid and Simultaneous determination of Cyclamate and Saccharine in food samples by HPLC with direct UV-detection

VICARIO, ANA; SAIDMAN, ELBIO; DEMARIA, CARLA; WANG, CHIEN C.; FELICI, EMILIANO; GOMEZ, MARÍA ROXANA

Rosario

Congreso; 4° Reunión de Ciencias Farmaceuticas RICiFa; 2016

Cyclamate and saccharine are artificial sweeteners that are commonly replacing sucrose in several beverages and foods. As they are non-caloric compounds, the use of artificial sweeteners has gained importance due to the special dietary requirements of people with diabetes. It is common to found a combination of both to compensate for each sweetener´s weaknesses and faults, such as bitterness and metallic taste.Up to date HPLC methodology combined with direct UV-Vis detection system has not been reported for simultaneous determination of cyclamate and saccharin. In this work, a simple, rapid and reliable HPLC methodology coupled to direct UV detection was proposed for sweetener samples analysis. All HPLC experiments were carried out on a Gilson HPLC system equipped with a Gilson 322 controller pump operating at 0.5 mL/min, a Rheodyne injector with a 20 μL sample loop, and a variable-wavelength UV-Vis DAD 170 Gilson detector measuring at 196 nm. The mobile phase consisted of buffer phosphate 20 mM, pH 2.5 (pump A) and acetonitrile-methanol 60:40 (pumo B), in a 75:25 proportion for pump A and pump B respectively. The analytical column used was a Restek Pinnacle II C18 (5 µm, 250 x 4.6 mm) (Bellefonte, PA, USA). Under the optimized experimental conditions, saccharine and cyclamate were successfully baseline separated in less than 10 minutes (tR saccharine=8.78 min; tR cyclamate= 9.51 min). Analytical parameters for saccharine were y=4.109x-1.106, R2=0.997 while for cyclamate were y=3.107x-6.104, R2=0.992. LOD and LOQ calculated were 4.10-4 mg/mL and 7.10-4 mg/mL, respectively, while for cyclamate were 0.0198 mg/mL and 0.0612 mg/mL respectively. Additionally, recovery rates were calculated in order to evaluate the applicability of the proposed methodology on real samples, achieving recoveries between 98-105% and 98-101% for saccharine and cyclamate, respectively. The developed methodology represents a simple, rapid and reliable tool that allows the simultaneous determination of saccharine and cyclamate in liquid artificial sweeteners. The results showed good applicability for monitoring of their contents in dietary products.