## 3 Results and Discussion  
### Identification of Polyphenolic Compounds in Hydromethanonic Extract of Salvia austriaca Haity Roots  
The hydromethanonic extract of _Salvia austriaca_ transformed roots was analyzed for polyphenolic compounds using UHPLC-PDA-ESI-MS (Figure 2 and Table 1). Three phenolic acids were identified by comparing their UV-Vis absorption spectra and MS/MS patterns with literature data [12]. The mass spectra of peak 1 revealed a pseudomolecular ion [M-H]\({}^{-}\) at \(m/z\) 179 and a major fragment ion at \(m/z\) 135, consistent with caffe acid [12]. Peak 2 exhibited a pseudomolecular ion [M-H]\({}^{-}\) at \(m/z\) 359 and three major fragment ions at \(m/z\) 197, 179, and 161, corresponding to rosmarinic acid [12]. Peak 3, with a pseudomolecular ion [M-H]\({}^{-}\) at \(m/z\) 493 and a major fragment ion at \(m/z\) 359, identified salvianic acid A [12]. The chemical structure of the identified phenolic acids is shown in Figure 3.  
solasodine biosynthesis in _Solanum khasianum_ hairy root cultures [15]. It is likely that light positively influenced caffe acid biosynthesis in _S. austriaca_ hairy roots, although rosmarinic and salvianolic acid A were also detected. While the presence of rosmarinic and salvianolic acids has been confirmed in _S. austriaca_[8], salvianolic acid A has not been previously reported in this species. Similar phenomena of novel derivative appearance have been noted in other in vitro cultures, such as new isomers, _iso_-sphaeralcic acid, and 8-methyl-iso-sphaeralcic acid, detected in _Sphaeralcea angustifolia_ transformed roots [16]. Effect of Cultivation Environment on Biomass and Phenolic Acid Production in S. austriaca Hairy Roots  
The morphology of _S. austriaca_ hairy root cultures after a 45-day growth period is illustrated in Figure 4.  
The duration of the culture was based on prior studies [17]. Roots cultured in SH liquid medium in shake flasks displayed typical growth patterns, oriented perpendicular to gravity. This pattern was also observed in the Plantform bioreactor (Figure 4). In contrast, roots grown in the mist-trickling bioreactor primarily grew in the direction of gravity (Figure 5). Similar growth patterns were observed in other nutrient-sprinkle bioreactors for transformed roots of _Salvia sclarea_[18], _Platyodon grandiflora_[19], and _Leonurus sibiricus_[20]. Figure 4: Forty-five-day-old _S. austriaca_ transformed roots grown in Plantform (**1**), mist-trickling (**2**) bioreactors, and the Erlenmeyer flask (**3**) in SH liquid medium (bar 1 cm). A bioreactor of similar design has previously been employed for the cultivation of _Salvia austriaca_ transformed roots [17]. However, the biomass parameters observed in that study were significantly lower than those recorded in the current investigation. This discrepancy may be attributed to excessive shear forces in the earlier system, which likely caused mechanical damage to the fragile root structures due to the sprinkler mechanism employed [10]. In response to this challenge, the present study introduced a newly designed bioreactor, developed by the primary author. To mitigate the adverse effects of shear forces on root growth, this bioreactor was equipped with a nozzle featuring an outlet diameter of 0.4 mm. This modification effectively reduced the particle size of the sprayed liquid medium, thereby significantly diminishing the unfavorable mechanical forces exerted on the roots. This improvement is likely responsible for the enhanced biomass accumulation and increased biosynthesis of secondary metabolites observed during cultivation in the newly proposed bioreactor design. Table 2 shows the _S. austriaca_ hairy root biomass data after a 45-day growth period. The mist-trickling bioreactor was more effective for biomass production, achieving 10.2 g dry biomass per liter of culture (155.4 g/L FW) after 45 days, compared to 9.7 g (149.29 g/L FW) from the Erlenmeyer flasks.