# Eco-Friendly Preservation of Pharaonic Wooden Artifacts using Natural Green Products
## 2 Materials and Methods
### Active Constituents of the Most Efficient Essential Oils  
The active constituents of the selected most efficient EOs were analyzed at the Laboratory of Medicinal and Aromatic Plants Research, National Research Centre, Dokki, Egypt, by a gas chromatography-mass spectrometry apparatus (TRACE GC Ultra Gas Chromatograph, THERMO Scientific Corp.,Waltham, MA, USA) coupled with a THERMO mass spectrometer detector (ISQ Single Quadrupole Mass Spectrometer). The GC-MS system was equipped with a TR5 MS column (30 m \(\times\) 0.32 mm i.d., 0.25 \(\mu\)m film thickness). The column temperature was kept at 60 \({}^{\circ}\)C for 5 min, then ramped at 5 \({}^{\circ}\)C/minute up to 200 \({}^{\circ}\)C, and finally kept for 10 min at that same temperature. A flame injector detector (FID) \begin{table}
\begin{tabular}{c c c c c} \hline \hline
**No.** & **Common Name** & **Latin Name** & **Family** & **Used Part** \\ \hline
1 & Einamon & _Cimnamomom cassia_ & _Lauraceae_ & Bark \\
2 & Eucalyptus & _Eucalyptus globulus_ & _Myrtaceae_ & Leaves \\
3 & Frankincense & _Boswellia carterii_ & _Burseraceae_ & Seeds \\
4 & Geranium & _Pelargointointa gracolens_ & _Gerasticaceae_ & Leaves \\
5 & Lavender & _Laroundula latifolia_ & _Lamiacane_ & Leaves \\
6 & Lemongrass & _Cymbopozion citratus_ & _Pouaceae_ & Leaves \\
7 & Menthe & _Menthe pipertia_ & _Lamiacane_ & Leaves \\
8 & Rosemary & _Rosmarinus officinalis_ & _Lamiacane_ & Leaves \\
9 & Tea tree & _Melaleuca alternifolia_ & _Myrtaceae_ & Leaves \\
10 & Thyme & _Thymus vulgaris_ & _Lamiacane_ & Leaves \\ \hline \hline \end{tabular}
\end{table} 
Table 2: Tested essential oils, plants and the parts from which they were obtained. \begin{table}
\begin{tabular}{c c c c c} \hline \hline
**No.** & **Common Name** & **Latin Name** & **Family** & **Used Part** \\ \hline
1 & Basil & _Ocimum basilicum_ & _Lamiacane_ & Leaves \\
2 & Eucalyptus & _Eucalyptus camalduncensis_ & _Myrtaceae_ & Leaves \\
3 & Henna & _Lawsonia inermis_ & _Lythracene_ & Leaves \\
4 & Melia & _Melia azadranch_ & _Meliaacue_ & Leaves \\
5 & Teak & _Tectona grandis_ & _Verbenaceae_ & Leaves \\ \hline \hline \end{tabular}
\end{table} 
Table 3: Plant extracts tested in this work and the parts from which they were obtained. with an operating temperature of 220 \({}^{\circ}\)C and an injector with a temperature of 180 \({}^{\circ}\)C was used for manual injection and with nitrogen as the gas carrier [40]. Effect of the Most Efficient Essential Oils on the Enzymatic Activities of the Highly Frequent Deterioration-Causing Fungal Species  
Activities of the different EOs on the amylase, cellulase, and protease of the most dominant deterioration-causing fungal species were determined. In particular, 100 mL of modified [PERSON]'s dox broth with 2% starch and 2% cellulose instead of sucrose and 1% casein instead of NaNO\({}_{3}\) were used. The reaction mixture was performed by adding 1 mL of 1% soluble starch in acetate buffer (0.02 M, pH 5.0) with 1 mL of the fungal filtrate and incubating for 15 min at 50 \({}^{\circ}\)C. The nihroyrin test was used to detect the hydrolysis of casein by protease, while the culture filtrate (1 mL) was mixed with 1 mL of 1% casein in acetate buffer (0.05 M, pH 5.6) as a substrate for 15 min at 37 \({}^{\circ}\)C; the reaction was terminated by adding 5% trichloroacetic acid (TCA) followed by a centrifugation at 3000 rpm for 10 min. Stock solutions of the EOs and plant extracts were prepared in DMSO and diluted with the broth to obtain different concentrations (0.125, 0.25, 0.5, 0.75, and 1 \(\mu\)L/mL) and added to a 250 mL conical flask followed by inoculation with a mycelia disc (10 mm) cut from the margin of the 7-day-old culture of the tested fungal species. The controls were prepared without the addition of EOs [41; 42; 43; 44]. The results are expressed as units per mL (U/mL). One unit is defined as the amount of enzyme which releases reducing sugars equivalent to 1 \(\upmu\)g of glucose.