*2.1. LC-MS Chemical Analysis (Phytochemical Analysis)*

Given that polyphenols are thought to be the primary bioactive chemicals in *Alchemilla* species, the polyphenolic profile has been thoroughly examined. According to our previous study conducted on the same extract, 23% of the polyphenolic components of *A. viridiflora* are present in the dry methanol extract [16]. Similarly, ellagitannins and flavonoids are two of the most abundant polyphenol classes in the sample used in this study. A total of 17 compounds were identified using LC-MS analysis (mass spectra and chromatograms are shown in Figures S1 and S2 of the Supplementary Materials, respectively) of *A. viridiflora* methanol extract with subsequent MS data processed and analyzed using MestReNova software (Table 1). Pedunculagin, tellimagrandin I, tellimagrandin II and galloyl-bishexahydroxydiphenoyl (HHDP) hexose constituted the major ellagitannin fraction of *A. viridiflora* extract while flavonoid fraction comprised quercetin, quercetin derivates and kaempferol glycosides. The recent chemical characterization of *A. viridiflora* extract provided by Radovic et al. (2022) is consistent with the present phytochemical analysis [16].


**Table 1.** Polyphenols identified in *A. viridiflora* methanol extract sample by LC-MS method.

Pedunculagin is a significant monomeric ellagitannin commonly found in *Alchemilla* species that has been related to various biological activities such as antitumor, antioxidant, gastroprotective, hepatoprotective, and anti-inflammatory properties [18]. Additionally, tellimagrandin I, a compound that was just recently identified as a constituent of *Alchemilla*

species, and brevifolin carboxylic acid, an *Alchemilla* ellagitannin product of hydrolysis that is also typically found in various pomegranate parts, are both recognized for their bioactivity and significant antiviral activity [16,19,20]. Compounds from the flavonoid class are equally important in terms of biological function. Numerous activities, including those related to the prevention of SARS-CoV-2, have been linked to quercetin, its derivatives and isorhamnetin [21].
