**2.2.3 HPLC analysis**

412 Macro to Nano Spectroscopy

Ba Bc Bg Bm Bp Bphy Bs Btr Bt

250 350

300

Btr 225, 250

Table 2. Wavelengths (nm) with statistically significant differences (p <0.05) among the species. *B. articulata* (Ba), *B. crispa* (Bc), *B. gaudichaudiana* (Bg), *B. microcephala* (Bm), *B. penningtonii* (Bp), *B. phyteumoides* (Bphy), *B. sagittalis* (Bs), *B. triangularis* (Btr) and *B. trimera*

A TLC analysis was initiated of the dichloromethane extracts of *B. articulata*, *B. gaudichaudiana* and *B. sagittalis* used to perform UV-Visible spectra using the mobile phase DCM: Hexane: MeOH (4:2:1) in order to complement the studies carried out by PCA of the spectrophotometric data and to find potential markers of the species that could not be

> 325 425

> 325 425

> 300 325 350

350 300

300 425

250 350

300 325

 225, 250, 300, 325 350

 225, 250 300, 325 350

225, 250 300, 325 350, 375

> 225 425

> 300 325 425

225, 250, 300, 325, 350, 375

> 250 350 375

> 300 325 375

250, 300 325,350 375

325, 350 375, 425

250

325

Bm 250

Bp 250

Bphy 250

Bs 225

325 350 375

250, 300, 325, 350 375

> 300 325 350

Ba 300

Bt

(Bt)

**2.2.2 TLC analysis** 

325

Bg 250

Bc 300

HPLC analysis was carried out on the same extracts as used for the studies with UV-Visible spectrophotometry and for TLC in *B. articulata*, *B. gaudichaudiana* and *B. sagittalis.* The chromatophotographic profiles showed the main peaks with the following retention times for *B. articulata*: 2.32, 3.00, 3.22 and 3.30 min; for *B. gaudichaudiana*: 2.26, 3.00, 3.12 and 3.30 min and for *B. sagittalis*: 2.26, 2.40, 3.00, 3.12 and 3.30 min. The retention time for the apigenin marker was 3.00 min and for genkwanin was 3.30 min. These peaks appear in all three species studied. In *B. gaudichaudiana* and *B. sagittalis* there is also a peak at 3.12 min, and this retention time corresponds to the luteolin marker (Figure 10).

Quality Control of Herbal Medicines with Spectrophotometry and Chemometric Techniques

– Application to *Baccharis* L. Species Belonging to Sect – Caulopterae DC. (Asteraceae) 415

Fig. 10. Original chromatograms obtained by HPLC of *B. articulat*a (Ba), *B. gaudichaudiana*

The growing tendency to use high quality, standardized plant extracts, with a guarantee of security and efficacy, is going to continue. Therefore, all efforts should be directed at researching the chemical-pharmalogical profiles of the extracts and combinations and in

(Bg) and *B. sagittalis* (Bs),. Compounds: 1, apigenin; 2, genkwanin; 3, luteolin.

**3. Conclusion** 

rationalizing their therapeutic applications.

Fig. 9. TLC of *B. articulata (Ba), B. gaudichaudiana (Bg)* and *B. sagittalis (Bs)*. Chromatograms were sprayed with NP-PEG and observed under UV 365 (A) or under UV 254 without chemical treatment (B). A, apigenin; G genkwanin; L: luteolin. Mobile phase: DCM: Hexane:MeOH (4:2:1)

Fig. 10. Original chromatograms obtained by HPLC of *B. articulat*a (Ba), *B. gaudichaudiana* (Bg) and *B. sagittalis* (Bs),. Compounds: 1, apigenin; 2, genkwanin; 3, luteolin.
