Table 4.

Summary of bioactive compound identification and characterization techniques used for E. alba.

Part of the Plant	Identification and Characterization	Operating Parameters	Bioactive Compounds	References
Whole plants	TLC	SP—silica gel 60, MP—chloroform–methanol, detecting agent—iron (III) chloride	Wedelolactone	[94]
	GC–MS	A GC–MS with an elite column, IV—2 μL with temperature range from 40 to 280 °C, carrier gas—helium, 1 mL/min	Various phytochemical compounds	[95]
Aerial parts	NMR	1H–NMR at 400 MHz and 13C–NMR at 101 MHz using dimethyl sulfoxide as a solvent	Luteolin, luteolin-7-O-glucoside, wedelolactone, and eclalbasaponin I,	[10]
	LC–MS	UHPLC–Q–TOF–MS UPLC—HSS T3 column, MP—acetonitrile and water (containing 0.1% formic acid), IV—5 μL, 0.3 mL/min at 30 °C. LC–QQQ–MS HPLC—C18 column, MP—acetonitrile and water (containing 0.1% formic acid), IV—1 μL, 0.5 mL/min at 35 °C.	Luteolin-7-O-β-D-glucopyranoside, luteolin, apigenin, ecliptasaponin A, C, and I,	[96]
			28-O-β-D-glucopyranoside, echinocystic acid, and 3-oxo-16α-hydroxy-olean-12-en-28-oic acid
	GC–MS	HP–5 MS capillary column (5% diphenyl, 95% dimethyl polysiloxane) at 70 eV, 50–600 amu of mass scan range, 1 mL/min, IV—0.1 μL with temperature range from 60 °C to 215 °C for 45 min	Fifty-nine volatile compounds, including α-pinene, caryophyllene, α-humulene, α-pinene, camphene, allo-aromadendrene, α-amorphene	[97]
	GC–MS	GC–HP–5MS capillary column at 70 eV, maximum temperature at 350 °C, helium as carrier gas, 1 mL/min, IV—1 μL	Heptadecane, n-hexadecanoic acid, pentadecane, and other 52 volatile compounds	[66]
	LC–MS, NMR	ESI–MS analyses—Triple Quad LC–MS instrument, HR–ESIMS analysis—Q–TOF mass spectrometer, NMR—a Bruker Avance DRX600 spectrometer, 2D 1H-1H COSY and HMBC NMR	Ecliprostins A–C	[11]
	FT–IR, LC–MS, NMR, UV	Varian 640-IR FT–IR spectrophotometer, Negative-ion HRESI–TOF–MS, Bruker 500 MHz NMR at 500 and 125 MHz, using tetramethylsilane (TMS), UV–Vis spectrometer	Ecliptasaponin A, 7-O-methylorobol-4′-O-β-D-glucopyranoside, 3-oxo-16α-hydroxy-olean-12-en-28-oic acid, 3′-hydroxybiochanin A, echinocystic acid, echinocystic acid 28-O-β-D-glucopyranoside, eclalbasaponin I and IV	[82]
	NMR, MS, IR, UV, GC–MS	1D, 2D NMR spectra and TOF LC–MS, FT–IR spectrometer, UV–Vis spectrophotometer, GC apparatus using an L-Chirasil-Val column	Three new triterpenes	[12]
Leaves	GC–MS	Elite-5MS (5% diphenyl/95% dimethyl polysiloxane), a capillary column, at 70 eV, IV—2 μL with temperature range from 110 to 280 °C, and the carrier gas—helium, 1 mL/min for 36 min	Methyl ester, methyl, methyl ester, pentadecanic aciddiethyl phthalate, glycine, c-sitosterol, eicosyl ester, and 10-octadeconic acid	[98]
	GC–MS	REX column, Temperature range 70–300 °C, helium—carrier gas, IV—2 μL	Glycine, hydrazine carboxyamide, garbamic acid, naphthoquinone, and other substances	[99]
	LC–MS	LC–ESI–QTOF–MS/MS—C18, 500 µL/min, IV—5 µL, MP—water + 0.1% formic acid and acetonitrile + 0.1% formic acid at 35 °C	Phenolic acids (gallic acid, protocatechuic acid, etc.), Flavonoids (luteolin, etc.), wedelolactone, triterpenoids, and phenolic aldehyde (protocatechualdehyde)	[22]
	HPTLC	Silica 60F 254 on aluminum sheet, 10 s/µL of syringe injection rate; IV—2 µL for plant extract and 1 µL for standards; MP—toluene–ethyl acetate–formic acid for 5 min at room temperature	Wedelolactone, luteolin, chlorophyll	[22]
	TLC, Q–TOF–MS, UV, NMR, FT–IR	Silica gel 60 F254 precoated plates, MP—ethyl acetate–toluene–formic acid–methanol, the spraying agent—NP-PEG, Micromass, Q–TOF–MS, UV–Vis spectrophotometer, Bruker AV-500 NMR spectrometer using DMSO as a solvent	Luteolin	[84]
	FT–IR	BRUKER IFS 66 model FT–IR spectrometer	Wedelolactone	[24]
	GC–MS	Elite-5MS (5% diphenyl/95% dimethyl polysiloxane), a capillary column, at 70 eV, IV—2 μL with temperature range from 110 to 280 °C, and the carrier gas—helium, 1 mL/min for 36 min	2-ethyl-2-methyl, 1-Heptatriacotanol, butyl octyl ester, Dodecanoic acid, Oleic acid, eicosyl ester, 9,19- Cyclocholestan-3-ol-7-one,4a-dimethly-[20R], 10- Octadecenoic acid, c-Sitosterol, methyl ester, 1,2 Benzenedicarboxylic acid, 10 methyl, methyl ester, Tridecanol	[100]
	SEM–EDS	20 kV, high vacuum mode	Elements including sodium, magnesium, potassium, calcium, copper, zinc, and iron	[24]
Stems	FT–IR	BRUKER IFS 66 model FT–IR spectrometer	Wedelolactone	[24]
	SEM–EDS	20 kV, high vacuum mode	Elements including sodium, magnesium, potassium, calcium, copper, zinc, and iron	[24]
Roots	SEM–EDS	20 kV, high vacuum mode	Elements including sodium, magnesium, potassium, calcium, copper, zinc, and iron	[24]
	FT–IR	BRUKER IFS 66 model FT–IR spectrometer	Wedelolactone	[24]