**2.9. Determination of amino acids**

Free amino acids (FAAs) in samples were determined using HPLC according to the method proposed by Rozan et al. (2000). A 20 μL aliquot of amino acid standard and digested sauce samples were transferred into vials and dried under vacuum. Then 20 μL of drying reagent containing methanol, water and triethylamine (ratio 2:2:1 v/v) was added. Then 20 μL of derivatizing reagent containing methanol, triethylamine, water and phenylisothiocyanate (PITC) (ratio 7:1:1:1 v/v) was added. The derivatized samples were then dissolved in 100 mL of buffer A that was used as mobile phase for HPLC. A Purospher® STAR RP-18e, 5 μm column was used with buffer A (0.1 M ammonium acetate, pH 6.5) and buffer B (0.1 M ammonium acetate containing acetonitrile and methanol, 44:46:10 v/v, pH 6.5) as mobile phase set for linier gradient at the flow rate of 1 mL/min. The injected sample volume was 20 μL and monitored at 254 nm of wavelength.

Potential of Fermented Sausage-Associated Lactic Acid Bacteria to Degrade Biogenic Amines During Storage 103

*sake*, were non-amine forming strains. The ability of AO exhibiting strains of LAB to degrade

**Figure 1.** a\* Value during ripening of Nham control at 25°C (), 30°C (), 37°C (▲) and Nham with

**Figure 2.** b\* Value during ripening of Nham control at 25°C (), 30°C (), 37°C (▲) and Nham with

and the a value of Nham control at 72 hours 37°C was higher than the other sample.

Fig. 1 showed a\* values represent red color of Nham during ripening time and temperature at 25°C, 30°C and 37°C, respectively. The results showed a value increased according to ripening

amine in vivo during sausage ripening was investigated.

starters (*L. plantarum* + *L. sake*) at 25°C (), 30°C (), 37°C ().

starters (*L. plantarum* + *L. sake*) at 25°C (), 30°C (), 37°C ().

## **2.10. Statistical analysis**

Data was analysed by one-way ANOVA and differences among treatment means were determined by Duncan's new multiple-range test.
