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In this study, we aim at establishing the first standard procedure of using commercial protease to collect fish protein hydrolysate (FPH) from the non-toxic pufferfish Lagocephalus wheeleri and set general specification of FPH as a functional food ingredient. Pufferfish Lagocephalus wheeleri was sorted and collected as by-caught from Vietnemease Northern and Central seas. General standard analyzing methods were used for sensory, nutritional, and chemical properties. The hydrolysis conditions were evaluated as a 1.2% mixture of Flavourzyme and Protamex proteases (1:1) at 55oC in 6 hours.  Qualified FPH was a bright yellowish color clear liquid with specialty fish smell and sweetness of amino acid with an aftertaste, total nitrogen 20.08 ± 0.25 g/l, Na.a/Nts ratio over 50%, with no TTXs detected. The hydrolysis conditions and processes were discussed and shown to be similar to other studies on different fish species. The benefit of making FPH using the materials from non-toxic pufferfish was also analyzed. Organization and control of accurate non-toxic pufferfish collection is the essential step to ensure a safe and highly commercialized product. The process established in this study can be upgraded into large-scale production of pufferfish FPH and allows further research for value-added products that are benefiting the Vietnamese fisheries industry.

Bui Thi Thu Hien