# Phytochemical Analysis of _Symphytum officinal_ Root Culture Extract
## 3 Results  
It was found that the studied extract samples contained a unique polymer--a derivative of caffe acid, which is a new class of natural polyether with 3-(3,4-dihydroxyphenyl) glyceric acid as a repeating unit. Figure 1 shows the UV spectrum (max absorption at 206; 285 nm with a 236 nm wing) of this polymer. Table 1 shows the results of spectrophotometric studies in the UV range of the key fractions of biologically active substances of extract samples of _Symphytum officinal_ root cultures obtained under various technological modes (4 h extraction). \begin{table}
\begin{tabular}{c c c c c} \hline \multirow{2}{*}{**Sample No. **} & \multirow{2}{*}{\ \begin{tabular}{c} **Temperature,** \\ **\({}^{\circ}\)C** \\ \end{tabular}  } & \multicolumn{2}{c}{ \begin{tabular}{c} **Volume Fraction** \\ **of Ethanol in the** \\ **Extractant, \%** \\ \end{tabular}  } & \multirow{2}{*}{**OD**} & \multirow{2}{*}{**m, mg/100 mL**} \\ \hline
1 & 30 & 30 & 0.0160 \(\pm\) 0.0001  & 58 \(\pm\) 0.1  \\
2 & 30 & 40 & 0.0230 \(\pm\) 0.0001  & 60 \(\pm\) 0.2  \\
3 & 30 & 50 & 0.0260 \(\pm\) 0.0001  & 75 \(\pm\) 0.1  \\
4 & 30 & 60 & 0.3562 \(\pm\) 0.0050  & 78 \(\pm\) 0.2  \\
5 & 30 & 70 & 0.2845 \(\pm\) 0.0030  & 73 \(\pm\) 0.1  \\
6 & 40 & 30 & 0.2118 \(\pm\) 0.0030  & 72 \(\pm\) 0.2  \\
7 & 40 & 40 & 0.2138 \(\pm\) 0.0030  & 160 \(\pm\) 0.1  \\
8 & 40 & 50 & 0.2652 \(\pm\) 0.0030  & 180 \(\pm\) 0.1  \\
9 & 40 & 60 & 0.1942 \(\pm\) 0.0030  & 240 \(\pm\) 0.1  \\
10 & 40 & 70 & 0.3827 \(\pm\) 0.0056  & 235 \(\pm\) 0.2  \\
11 & 50 & 30 & 0.2918 \(\pm\) 0.0050  & 230 \(\pm\) 0.1  \\
12 & 50 & 40 & 0.3315 \(\pm\) 0.0056  & 390 \(\pm\) 0.2  \\
13 & 50 & 50 & 0.2180 \(\pm\) 0.0050  & 370 \(\pm\) 0.1  \\
14 & 50 & 60 & 0.4490 \(\pm\) 0.0059  & 330 \(\pm\) 0.1  \\
15 & 50 & 70 & 0.3680 \(\pm\) 0.0056  & 332 \(\pm\) 0.1  \\
16 & 60 & 30 & 0.1290 \(\pm\) 0.0010  & 250 \(\pm\) 0.2 [FOOTNOTE:2]Footnote 2: Data presented as a mean \(\pm\) SD (\(\pi\) = 3). Values in columns followed by the same letter do not differ significantly (\(p\) \(>\) 0.0 Figure 2 presents an HPLC chromatogram of the _Symphytum officinal_ extract samples. The components and their retention times, found under HPLC conditions, are presented in Tables 2 and 3. \begin{table}
\begin{tabular}{c c c c} \hline \hline
**Peak No.** & **Retention Time, min** & **Component** &
\begin{tabular}{c} **Quantitative** \\ **Content***, \#g/mL** \\ \end{tabular} \\ \hline
1 & 6.164 & m-methoxybenzoic acid & 30.05 \(\pm\) 0.49 \\
2 & 6.502 & 5-caffeoylquinic acid & 14.73 \(\pm\) 0.72 \\
3 & 7.300 & caffeic acid & 25.40 \(\pm\) 0.60 \\
4 & 7.811 & 3-caffeoylquinic acid & 12.57 \(\pm\) 0.62 \\
5 & 8.528 & rosmarinic acid & 2.03 \(\pm\) 0.05 \\
6 & 9.620 & 1-caffeoylquinic acid & 3.12 \(\pm\) 0.20 \\
7 & 11.350 & coumarin & 7.32 \(\pm\) 0.41 \\
8 & 13.426 & lithospermic acid & 11.27 \(\pm\) 0.52 \\
9 & 14.270 & salicylic acid & 9.25 \(\pm\) 0.50 \\
10 & 15.030 & m-hydroxybenzoic acid & 1.97 \(\pm\) 0.20 \\ \hline \hline \end{tabular}  
* average of three measurements.  
\end{table} 
Table 2: Component composition of the aqueous-alcoholic extract of _Symphytum officinal_, containing type I polymer, according to HPLC data. Figure 2: HPLC chromatogram of the _Symphytum officinal_ extract samples containing (**a**) type I and (**b**) type II of the recovered polymer: peak 1—m-methoxybenzoic acid; 2—5-caffeoylquinic acid; 3—caffeic acid; 4—3-caffeoylquinic acid; 5—rosmarinic acid; 6—1-caffeoylquinic acid; 7—cournarin; 8—lithospermic acid; 9—salicylic acid; 10—m-hydroxybenzoic acid; and 11—3_A_-dimethoxybenzoic acid (unnumbered peaks of the chromatogram belong to unidentified substances). The results of the chromatographic analysis are presented in Figure 3.  
9-Angeloylretronecin (IUPAC ((7R,8R)-7-hydroxy-5,6,7,8-tetrahydro-3H-pyrrolizin-1-yl)methyl (Z)-2-methylbut-2-enoate) and 7-Angeloylheliotridine (IUPAC ((1S,8R)-7-(hydro-xymethyl)-2,3,5,8-tetrahydro-1H-pyrrolizin-1-yl) (Z)-2-methylbut-2-enoate) were found in comfrey extract samples.