Methanolic Synthesis

Based on Gursale et al. method [25] methanolic extraction was applied to completely extract and quantify the amount of different alkenylbenzenes present in the PFS samples. The extraction was performed by adding 50 ml methanol to 0.5 g of PFS followed by sonication for 15 minutes. Upon sonication the extract was filtered using a 0.45 um syringe filter and the filtrate was stored at -20 °C until Ultra Performance Liquid Chromatography (UPLC) analysis.

UPLC analysis.
For analysis of the methanolic PFS extracts the samples were analysed both in undiluted form and also diluted 10 times with acetonitrile because the amount of the alkenylbenzenes was expected to be high. UPLC analysis was performed using a Waters (Waters, Milford, MA) Acquity …show more content…
To this end, PFS sample number four (Table 1) was spiked with apiol, myristicin and estragole. The spiked sample was analyzed by the same procedure as described above.

Extraction procedure linearity.
To determine the linearity of the extraction method, solutions of sample number four were prepared as follow: (0.5g/200 ml, 0.5 g/100 ml, 0.5g/50 ml, 0.5/25 ml, 1g/25ml. The results was shown to be linear with the amount of 0.5 g PFS in 25 ml methanol. (data not shown)

Risk assessment of estimated intake of alkenylbenzenes found in parsley or dill PFS.
Risk assessment will be performed using the MOE approach. The MOE values were calculated using EDI values calculated based on Aljlouni at el.[26]; first the MOE values were calculated for the individual alkenylbenzenes using their respective BMDL10 values, then the MOE values were calculated using a combined exposure assessment assuming equal potency of all alkenylbenzenes and the BMDL10 for the major alkenylbenzene in the mixture and finally MOE values were calculated based on the Toxic Equivalency (TEQ) approach calculating the combined exposure in safrole equivalents and using the BMDL10 of safrole through defining Toxic Equivalency Factors (TEFs) for the apiol, myristicin and estragole. The TEF values of apiol, myristicin and estragole are 0.30, 0.66 and 2.16 respectively [26]. BMDL10 values were obtained from literature [18, 19,

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