LIQUID CHROMATOGRAPHIC ANALYSIS OF FREE AMINO ACIDS IN AMERICAN COCKROACH ( Periplaneta americana )

Insects could serve as an alternative amino acids source with less environmental impact and low production cost. In this study, phenylisothiocianate (PITC) pre–column derivatization and high-pressure liquid chromatographic (HPLC) method combined with solid phase extraction (SPE) was developed and used to quantify free amino acids present in American cockroach. Powdered cockroach sample were extracted in water at 90 C for 20 min, pre-purified on silica-based cationic exchange (SCX) cartridges, derivatized and analyzed for the free amino acid compositions. The free amino acids derivatives were separated on Agilent Zorbax Extend-C18 (4.6 x 250 mm, 5 μm) HPLC column at 33 ± 0.1 C with diode array detection (DAD) set at 254 nm. A total analysis time of 30 min was used to separate 15 amino acids including nine essential and six non-essential ones with gradient elution at a flow rate of 1.0 mL/min. The results showed linear calibration range from 50 to 150 pmol/uL for all the free amino acids. The sensitivity expressed as limits of quantification (LOQ) and limit of detection (LOD); repeatability and reproducibility precisions expressed as relative standard deviation (RSDs) percentages for all amino acids characterized by the following ranges:2.6 to 35.5 pmol/μL; 0.9 to 11.7 pmol/μL; 1.0 to 10.5 %; 6.8% to 51.9%. The optimized and validated analytical method was applied for the determination of free amino acids in American cockroach. The free amino acids concentrations ranged from His, 0.43 mg/g to Pro, 311.2 mg/g and the recovery ranged from Thr, 82.9 % to Glu and Gly, 100 %. The free amino acids concentrations were in the order Pro > Thr > Leu > Glu > Ile > Lys > Met > Val > Asp > Ser > Try > Gly > Arg > His. The concentration of Pro was 5 – 100 LIQUID CHROMATOGRAPHIC ANALYSIS OF FREE AMINO ACIDS IN AMERICAN COCKROACH (Periplaneta americana) SDRP Journal of Food Science & Technology (ISSN: 2472-6419) DOI: 10.25177/JFST.4.1.RA.434 Research

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