#### 2.2.1. Bufadienolides  
Bufadienolides are C-24 steroids with a pyranone ring at C-17\(\beta\) that are naturally present in plants and mammalian animals, specifically toads and snakes [19]. Bufadienolides were found more abundant in WS than RS as revealed from the inspection of the chromatograms of both WS and RS at retention time \"Rt.\" from 5 to 13 min. A series of different classes of bufadienolides were identified in WS and RS and included hydroxy-oxobufadienolide with a C-19 aldehyde group, i.e., bufotalidin, and hydroxybufaenolides with a C-19 methyl group, including scillarenin and scillphosidin. Notably, identified bufadienolides structures were illustrated in Figure 3.  
\begin{table}
\begin{tabular}{c c c c c c c c c c c} \hline \hline
**Code** & **tg (ms)** & **UV** & **Elemental** & **Empar ppm** & **Exact Maps** & **Lospital** & **Lospital** &  
Hydrooxyoxobufaenolide  
Hydrooxyoxobufaenolides were detected in multiple peaks L53, L37, L49, while L36, L63, L67 and L68 were annotated as glycosides of hydroxyoxobufaenolides [20; 21; 22]. L36, L63, L67, and L68 were identified as hydroxyoxobufaenolide glycosides based on the loss of \(-\)162 or 146 amu. in L36 desacetylscillirosidin-O-thevetooside (\(m\)/z 577, (C\({}_{31}\)H\({}_{44}\)O\({}_{10}\))\({}^{+}\)) (Figure S1A), L63 Hydrooxysciliroside (\(m\)/z 637, (C\({}_{32}\)H\({}_{44}\)O\({}_{13}\))\({}^{+}\)) (Figure S1B) [23], L67 m/z 579 bufatalidin-O-hexoside, and L68 \(m\)/z 545 (C\({}_{30}\)H\({}_{41}\)O\({}_{9}\))\({}^{+}\) [M+H]\({}^{+}\) scilligiaucosidin-O-rhannoside (Figure S1C).  
Identified hydroxygufaenolides are subdivided by the number of hydroxyl groups to mono, di, tri and tetra hydroxyl derivatives and were identified in both WS and RS as listed in Table 2.  
The L64 peak \(m\)/z 367 (C\({}_{24}\)H\({}_{31}\)O\({}_{3}\))\({}^{+}\) [M+H]\({}^{+}\) was detected in both WS and RS as monohydroxybufa-tetraenolide \"Scillaridin A\" [24; 25; 26] as an example of a monohydroybufaenolide, illustrated in Figure S1D.  
Dihydroxygufaenolides were detected in several peaks including L82 \(m\)/z 387 (C\({}_{24}\)H\({}_{35}\)O\({}_{4}\))\({}^{+}\) [M+H]\({}^{+}\) dihydroxybufadienolide \"bufalin\" [27] and L70 \(m\)/z 385 (C\({}_{24}\)H\({}_{33}\)O\({}_{4}\))\({}^{+}\) [M+H]\({}^{+}\) dihydroxy-bufa-trienolide \"Scillarenin\" [28; 29; 30]. Glycosic conjugates were observed in L91 \(m\)/z 531, showing a loss of rhamnose (\(-\)146 amu) (Figure S1E) and L86 \(m\)/z 693 (C\({}_{36}\)H\({}_{53}\)O\({}_{13}\))\({}^{+}\) [M+H]\({}^{+}\) with a loss of hexose (\(-\)162 amu) (Figure S1F). L91 and L86 annotated as scillarenin-O-rhannoside \"Proscillaridin A\" [11; 31; 32; 33] and prosci  
Figure 3: Chemical structure of bufadienolides in both RS and WS detected using UPLC/MS.  
Acylated bufaenolides were detected in L90 at _m_/z 607 (C\({}_{32}\)H\({}_{47}\)O\({}_{11}\))\({}^{+}\) [M+H]\({}^{+}\) and annotated as acetyl-scilliphaeoside. L39 at _m_/z 403 (C\({}_{24}\)H\({}_{35}\)O\({}_{5}\))\({}^{+}\) [M+H]\({}^{+}\) annotated as trihydroxybufa-dienolide \"gamabufotalin\" [29; 34; 35] (Figure S1G).  
Other acetylated tetrahydroxybufaenolide were detected in peak L54 at _m_/z 477, (C\({}_{26}\)H\({}_{37}\)O\({}_{8}\))\({}^{+}\) (M+2H)\({}^{+}\) identified as hydroxyscillirosidin with a loss of acetyl group likely at _C_-16 [26; 36; 37] (Figure S1H).