ANALYTICAL METHOD VALIDATION OF BENZENE USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY IN BEVERAGE CONTAINING SODIUM BENZOATE AND ASCORBIC ACID
Abstract: Several countries
reported discovering benzene in beverages containing benzoic acid and ascorbic
acid. Benzoic acid decarboxylation by ascorbic acid will form benzene. American
Beverage Association (ABA) recommended the use of accelerated testing to test
benzene in beverages. High Performance Liquid Chromatography (HPLC) is one of
several methods to analyze benzene in a wide variety of samples, but there is
no much information provided regarding the validation of analysis method of
benzene. Therefore, developing analysis method of benzene and its validation
becomes a current need. The HPLC system consists of Hitachi L-2130 pump, sample
injector with 20 µL sample loop, and UV detector L-2420 operating at 205 nm.
The analytical column is a LiChrosorb Phenomenex RP-18 (250 x 4 mm, 10 µm, 100
Ǻ), the mobile phase is acetonitrile:aquabidest (60:40 v/v) and pumped at a
flow rate of 0,8 mL/min.Benzene separated from the matrix and follows the
validation requirements. The developed analytical method showed that resolution
was 8.37, r = 0.995 with LOD and LOQ 6.52 ppb and 19.75 ppb, with a precise of
≤11% and recovery of 80-110%. Accelerated testing indicated that benzene levels
increased with increasing of the temperature. Beverages containing 400 mg/mL of
ascorbic acid and benzoic acid formed benzene which was detected as 699.38 ppb
at 25 oC, 799.61 ppb at 40 oC, and 808.94 ppb at 60 oC in 48 hours. In
conclusion, the method was fully validated and can be utilized to analyze
benzene in beverages with the accelerated testing at 60 oC in 48 hours, so that
benefits the producers and consumers in the end.
Penulis: Melania Perwitasari,
Endang Lukitaningsih, Sudibyo Martono
Kode Jurnal: jpfarmasidd140400
