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Brewing Chemistry Analysis –                                                                                              Packaging Testing for Beer

                    N-Nitrosamines



                    Most N-nitrosamines are known to be carcinogenic and   amount of nitrosamines in beer must be controlled to the           When beer is not ready for direct consumption, then there must   result in injury.
                    mutagenic. Consumption of nitrosamines, e.g.,      acceptable levels, typically at 0.2 ~ 5.0 ppb depending on the         be some type of packaging such as cans, bottles, or kegs. For   The Shimadzu EZ-X tensile testing system tests force versus
                    N-nitrosodimethylamine (NDMA), was reported to be a cause of   country. The main N-nitrosamine that is monitored in malt and   cans, they usually have a pull-tab. The question becomes how   stroke distance according to standard method JIS S 0022:2001.
                    gastric cancer, liver cancer, glioma and blood disorder.   beer is NDMA.                                                  easy or hard should the pull-tab be to open. If the pull-tab   The test requires to hold the can by hand; however, in the
                    N-nitrosamines can be formed by a reaction between amine and   Shown is a GC-MS/MS method using Multiple Reaction         opens too easily, then there is a possibility of leakage during   example below, a fixing jig was used to hold the can in position
                    nitrite under heating conditions in food processing. They have   Monitoring (MRM) mode for simultaneous detection and     shipping and handling. If the pull-tab is too hard to open, then   for more accurate testing results. A string is hooked on the
                    been found in malt-derived beverages like beer at trace levels.   quantification of six N-nitrosamines, including NDMA, for   the tab may break off before the can is opened. This may lead   pull-tab and it is pulled straight up at a speed of 500 mm/min.*
                    The presence of NDMA in malt and beer was first reported in   enhanced selectivity and sensitivity from the potential matrix   to people trying to puncture the can with a “tool” which can
                    1974. Its concentration in malt depends on the drying   interferences in beer samples*.
                    techniques used. According to US and EU regulations, the

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                                                                                                                                                                                                             Made-in-Japan product
                         (x100)        (x100)        (x100)                                                                                                                                           30     Made-in-Japan product (with large opening)
                         74.10>42.00   102.10>85.10   3.0  158.20>99.10                                                                                                                                      Made-in-Mexico product
                      2.25 74.10>30.00  102.10>56.00  158.20>141.20                                                                                                                                          Made-in-Holland product
                                     2.5
                      2.00                         2.5                                                                                                                                                25     Made-in-Belgian product
                      1.75           2.0
                                                   2.0
                      1.50
                                     1.5                                                                                                                                                              20
                      1.25                         1.5                                                                                                                                              Test force [N]
                      1.00           1.0
                                                   1.0                                                                                                                                                15
                      0.75
                                     0.5
                            6.7  6.8       7.3  7.4     9.4  9.5
                            NDMA          NDEA           NDBA                                                                                                                                         10
                         (x100)        (x100)        (x100)
                       1.3  114.10>97.10   100.10>70.00   116.10>86.00                                                                                                                                 5
                                           100.10>55.00
                                                     116.10>56.00
                                     6.0           4.0
                       1.2
                                                   3.5
                       1.1                                                                                                                                                                             0
                                     5.0           3.0
                       1.0                                                                                                                                                                              0    5    10   15    20   25    30   35
                       0.9           4.0           2.5                                                                                                                                                                   Stroke [mm]
                       0.8                         2.0                                                                                             Tensile test on pull-tab on beer can containers with EZ-X  Relationship between the Force and Stroke
                       0.7           3.0           1.5
                       0.6                         1.0
                       0.5           2.0           0.5
                            9.7  9.8       9.9  10.0  10.1  10.2
                            NPIP          NPYP           NMOP                                                                                               Sample                   Maximum Opening Strength [N]
                               MRM transitions of six N-Nitrosamines at            Shimadzu GCMS-TQ8040 triple                                   Canned beer (made in Japan)                 26.8
                               0.1 ppb in Beer by GC-MS/MS                         quadrupole mass Spectrometer                                  Canned beer (made in Japan, with a large lip)  30.8
                                                                                                                                                 Canned beer (made in Mexico)                26.5
                                                                                                                                                 Canned beer (made in Holland)               27.2
                                                                                                                                                 Canned beer (made in Belgium)               22.4
                                                                                                                                              Force required to open beer from various countries*

















                    *ASMS 2013 Poster TP738: “Method Development for Trace Level Detection of N-Nitrosamines in Beer by GC-MS/MS”             *Source: Application Data Sheet #25: “Tensile Test on Pull-Tab Containers (Reference Standard: JIS S 0022:2001)”

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