## 3 Results and Discussion  
### Total Phenolic Content  
The use of an environmentally friendly, modern extraction technique (SFE) allowed obtaining pure fractions collected at different temperatures. Supercritical CO\({}_{2}\) is a non-polar solvent, but its polarity can be modified by temperature and pressure variations. Polyphenols with high molecular mass are hardly soluble in pure CO\({}_{2}\), but their solubility can be enhanced by adding a polar co-solvent or increasing pressure [26]. In our research, methanol commonly used for extraction and high pressure (250 bar) were used. Testing the antioxidant activity allowed us to check the differences between fractions obtained at different temperatures. The TPCs of the extracts obtained from dry _Laminum album_ flower by SC-CO\({}_{2}\) with methanol as co-solvent were determined quantitatively using a spectrophotometric method with Folin-Ciocalteu reagent as the most and commonly used method for total phenols estimation. A calibration curve was established with gallic acid as standard at different concentrations. The results were expressed in mg of gallic acid equivalent (GAE)/g of extract. All measurements were taken in triplicate, and mean values were calculated. The results of determination of the polyphenols are presented in Table 1.  
As shown in Table 1, the results varied from 234.17 to 650.17 mg GAE/g of extract for the studied conditions, which represent a variation of approximately three folds. The extract at 50 \({}^{\circ}\)C showed the highest TPCs of 650.17 mg GAE/g of extract, followed by 60 \({}^{\circ}\)C, which showed 418.50 mg GAE/g of extract, and 40 \({}^{\circ}\)C showed the lowest value of 234.17 mg GAE/g of extract. All studied conditions showed a considerable amount of total phenols and they were significantly different as expected due to the effect of temperature on the density and selectivity of the solvent during SC-CO\({}_{2}\). However, a degradation effect may be suspected between 50 \({}^{\circ}\)C and 60 \({}^{\circ}\)C at 250 bar during the extraction process, which has caused a decrease in TPCs.  
The total phenolic content obtained in this work is not directly comparable to other studies of _Laminum album_ extracts because the extraction methods are different. In the previously published papers, most of them have used solvent extraction methods to extract _Laminum album_. [PERSON] and [PERSON] (2006) [27] reported 192 mg GAE/g of extract of TPC in methanolic extract of _Laminum album_, [PERSON] et al. (2012) [5] reported 500.7 \(\pm\) 50.0 mg/g of extract of phenolic compounds in the ethanolic extract of _L. album_[5], also other species of _Laminum_ have been reported to have high content of polyphenols [7; 27; 28] and methanol have been reported as a good solvent [28]. The high TPC value obtained for our extract (especially for the fraction at 50 \({}^{\circ}\)C) compared to the research of other authors indicates a very effective extraction using the SFE technique.  
\begin{table}
\begin{tabular}{l c} \hline \hline
**Extraction Conditions** & **TPC (mg GAE/g of Extract)** \\ \hline
40 \({}^{\circ}\)C & 234.17 \({}^{\mathrm{a}}\)\(\pm\) 2.48 \\
50 \({}^{\circ}\)C & 650.17 \({}^{\mathrm{b}}\)\(\pm\) 3.86 \\
60 \({}^{\circ}\)C & 418.50 \({}^{\mathrm{c}}\)\(\pm\) 2.92 \\ \hline \hline \end{tabular} Values are mean \(\pm\) s.d., \(n\) = 3, values with the same lowercase letters in the same column indicate no significant difference at level of 5\(\%\) (\(p\) < 0.05).  
\end{table}
Table 1: Total phenolic content of _Laminum album_ flower for extracts obtained at different variants of extraction.