# Microplastics' Detection in Honey: Development of Protocols in a Simulation
## 3 Results and Discussion
### Analysis of Mass Spectrometry and Results  
Confirming the PET masses (MH\({}^{+}\)) to the literature [30], fragments with \(m/z\) 167.0337 and 211.0599 corresponding to TPA and MHT, respectively, were found in D_C1, D_C2, and D_Final, and in the indicator sample with PET MPs and deionized water (MPs_H2O), by the ESI-MS technique using the LTQ-Orbitrap XL ETD mass spectrometer. As is presented in Table 3, the fragments did not exist in the corresponding control samples with honey (C_C1, C_C2, and C_Final), which was the most essential MS result. The common existence of the fragment ion 167 in the solvents does not disturb the identification of this fragment ion as PET ion in the samples. In the supernatants, these fragments were not identified as PET fragments because \"z\" was not determined from the analysis of Xcalibur software. Additionally, based on the reference table (Common Background Contamination Ions in Mass Spectrometry, Fisher Chemical, Ottawa, ON, Canada), there was no contamination in the samples. In Figure 10, the two fragments are presented at specific time points from the MPs_H2O samples. Additional spectra are presented in the Supplementary Section Figures S3-S9. Our result reveals that PET fragments can be detected in diluted honey solutions using the ESI-MS technique with the LTQ-Orbitrap XL ETD mass spectrometer simply by injecting the PET NP samples directly into the MS detector.  
\begin{table}
\begin{tabular}{c c c c c c c c} \hline \multicolumn{6}{c}{**First Extraction Phase**} & \multicolumn{4}{c}{**Second Extraction Phase**} \\ \hline  & \multicolumn{2}{c}{**Optical Mi-**} & \multicolumn{2}{c}{**Optical Mi-**} & \multicolumn{2}{c}{**Optical Microscopy/Raman**} & \multicolumn{2}{c}{**Imagej**} & \multicolumn{2}{c}{**MP Dimensions and Quantity Notes**} \\  & \multicolumn{2}{c}{**Mapping**} & \multicolumn{2}{c}{**Mapping**} & \multicolumn{2}{c}{**Mapping**} & \multicolumn{2}{c}{**Imagej**} & \multicolumn{2}{c}{**MP Dimensions and Quantity Notes**} \\ \hline \multirow{2}{*}{**Phase**} & \multirow{2}{*}{**MP Dimension**} & \multirow{2}{*}{**MP Count**} & \multicolumn{2}{c}{**Average**} & \multirow{2}{*}{**MP Dimension**} & \multirow{2}{*}{**MP Count**} & \multicolumn{2}{c}{**Average**} & \multirow{2}{*}{**Dimension**} \\  & & & **(um?)** & & & & **(um?)** \\ \hline D\_Y **(1,2)** & 1-20 μm & 54 & 12 & 1-20 μm & 41 & 19 & Many small and medium MPs \\ \hline D\_YC **(1,2)** & - & - & - & 18-50 μm & 27 & 28 & Medium MPs \\ \hline D\_YC **(1,2)** & - & - & - & A few 2-3 μm & \multirow{2}{*}{74} & First extraction \(\rightarrow\) Many small and medium MPs \\ \hline \multirow{2}{*}{D\_C **(1,2)**} & \multirow{2}{*}{1-30 μm} & \multirow{2}{*}{111} & \multirow{2}{*}{30} & Many 18-25 μm & \multirow{2}{*}{74} & \multirow{2}{*}{25} & \multirow{2}{*}{Medium MPs} \\  & & & & 100 μm clusters & & & & \\  & & & & Some 10 μm & & & & \\ \hline \multirow{2}{*}{**Sediment of**} & \multirow{2}{*}{No MPs \(<\) 8-74 μm} & \multirow{2}{*}{No red map} & \multirow{2}{*}{No red map} & \multirow{2}{*}{75 μm and \(>\)100 μm clusters} & \multirow{2}{*}{No red map} & \multirow{2}{*}{No red map} & First extraction \(\rightarrow\) Large MPs \\  & & & & Some 5 μm & & & Second extraction \(\rightarrow\) Large Clusters \(>\) \\ \hline D\_Final & & & & 3, 5, 10, 20 μm & \multirow{2}{*}{121} & \multirow{2}{*}{29} & The most MPs found. MPs \(<\) 1–2 μm \\  & & & & 55-75 μm & & & not found \\ \hline \end{tabular}
\end{table}
Table 2: Statistics on MP particle dimensions due to the density separation phases with pentane. All particles have been counted in the same field of view (740 \(\times\) 590 μm).