Table 1.

Linear diterpenes extracted from B. bifurcata, and the respective pretreatment, extraction, purification/fractionation and identification/analysis methodologies used, and time of collection/geographical origin.

Extraction Methodology	EY a	Sample Pretreatment	Fractionation/Purification	Identification/Analysis	Compound (Content b)	Time of Collection/Geographical. Origin	Ref
EtOAc S–L extraction	1.70	Freeze-dried	Chromatography (on Silica gel) eluted with EtOAc-n-heptane (4:1) and monitored at 270 nm; Bio-guided fractionation: chromatography (on a Lobar RP-8 column) with EtOH-H2O; Fr. purification with MeCN-H2O or EtOAc-CHCl3.	(1H and 13C) NMR, MS	9 (0.06)	July to August/Roscoff, Brittany, France	[28]
					15 (0.07)
					18 (0.11)
					19 (0.03)
					20 (0.02)
EtOAc S–L extraction	N.D.	Freeze-dried	LC using Silica gel with a solvent gradient from DCM to DCM-MeOH (80:20); CC (on Silica gel) with solvent gradient DCM-MeOH (100:0 to 97:3) HPLC (RP18 column) with a gradient elution (from H2O-MeCN (60:40), H2O-MeCN (80:20) to MeOH)	HRESIMS, 1D and 2D NMR	10	September/Roscoff, Brittany, France	[30]
					15
					18
					21
					27
					31
					32
					33
					34
					41
EtOAc S–L extraction, at RT	4.80	Freeze-dried and ground	Successive flash and CC (on Silica gel), eluting with cyclohexane—EtOAc mixture; Bio-guided fractionation: CC (on Silica gel), with cyclohexane-EtOAc	HREIMS, (1H and 13C) NMR	15 (0.004)	November to September/Basse-Normandie, France	[21]
EtOAc S–L extraction, at RT	N.D.	Freeze-dried and ground	Flash chromatography eluting with a H2O-MeOH mixture of increasing polarity (95:5–0:100 in 30 minutes)	2D NMR, HPLC-DAD-MS-SPE-NMR	15	June/Cap Lévi, English Channel, France	[22]
EtOAc S–L extraction, at RT	4.8	Freeze-dried and ground	CC (on Silica gel) eluted with a solvent gradient from DCM to DCM-MeOH (80:20) UV-ESI-DAD-HPLC (using a Kromasil RP18 column) eluted with a solvent gradient H2O-MeCN-MeOH	(1H and 13C) NMR, HRESI(+)MS	35 (0.002)	September/Roscoff, Brittany, France	[34]
					36 (0.001)
					44 (0.0004)
CHCl3 S–L extraction	N.D.		Normal and reverse phase flash CC and RP18 HPLC	1D and 2D NMR, HRMS	9	May/Kilkee, County Clare of Ireland	[48]
					10
CHCl3-EtOH S–L extraction	1.52	Freeze-dried and ground	Partitioning between H2O and Et2O; Et2O-soluble material: CC (on Silica gel) eluted with hexane-EtOAc (2:3); HPLC (EtOAc–isooctane, 2:3), with RI monitoring	IR, (1H and 13C) NMR, EIMS	1 (0.86)	-/Atlantic coast Morocco	[45]
CHCl3-MeOH S–L extraction, at RT	5.53	Shade-dried and ground	Partitioning in the mixture MeOH-isooctane (1:1); MeOH phase: dissolved in the mixture MeOH-CHCl3-H2O (4:3:1); Organic phase: CC (on Silica gel) eluted with a solvent gradient from isooctane to EtOAc and then from EtOAc to MeOH.	HREIMS, (1H and 13C) NMR	1 (0.37)	December/Oualidia, Morocco	[44]
					6 (0.01)
					7 (0.004)
CHCl3- MeOH S–L extraction; Et2O S–L extraction of aqueous phase	1.95 g ext	Freshly collected	Open CC (on Silica gel) eluted with hexane to EtOAc (2:3); HPLC (eluent EtOAc-isooctane, 2:3).	IR, UV, (1H and 13C) NMR, EIMS, HRMS	2 (0.17)	-/Morocco	[47]
					40 (0.02)
CHCl3-MeOH S–L extraction	2.63	Shade-dried	Partitioning in the mixture MeOH-isooctane (1:1); MeOH extract: dissolution in the mixture MeOH-CHCl3-H2O (4:3:1); Organic extract: fractionation on silica gel column eluted with EtOAc and EtOAc-MeOH (98:2 and 95:5); HPLC on C-18 reversed-phase column, eluting with MeCN-H2O (1:1 and/or 2:3).	HRMS, IR, (1H and 13C) NMR	8	-/Oualidia, Morocco	[49]
					43
CHCl3-MeOH S–L extraction, at RT CHCl3-H2O L–L extraction	6.92	Air-dried and ground	MeOH extract: defatting with n-hexane; CC (on Silica gel) using mixtures of CHCl3-MeOH (from CHCl3 to CHCl3-MeOH, 1:1).	HPLC	9	July/Quiberon, Brittany, France	[37]
					13
					15
					21
					25
					26
					27
					28
					29
					30
					39
CHCl3- MeOH S–L extraction, at RT	3.11	Shade-dried and ground	CC (on Silica gel) eluted with a solvent gradient from cyclohexane to EtOAc and then from EtOAc to MeOH; HPLC on an analytical C-18 reverse-phase column (eluent, MeCN-H2O), with RI monitoring	HREIMS, IR, (1H and 13C) NMR	9 (0.15)	July/Quiberon, Brittany, France	[24]
					13 (0.0001)
					15 (0.40)
					21 (0.01)
					25 (0.0001)
					26 (0.0003)
					27 (0.0002)
					28 (0.0001)
					29 (0.0005)
					30 (0.0005)
					39 (0.008)
					47 (0.01)
Et2O S–L extraction, at RT	1.65	Freeze-dried and powder	Partitioning between H2O and Et2O; Et2O-soluble material: CC (on Silica gel) eluted with a solvent gradient from hexane to EtOAc; HPLC (eluent, EtOAc–isooctane), with RI monitoring	HRMS, EIMS, IR, (1H and 13C) NMR	1 (0.33–0.34)	January to December/Atlantic coast of Morocco	[43]
					2 (0.07–0.10)
					9 (0.10–0.12; 0.36–0.38)
					15 (0.29–0.32)
					40 (0.03)
					41 (0.01;0.04–0.05)
Et2O S–L extraction	2.94	Air-dried and ground	CC (on Silica gel) eluted with a gradient from isooctane to EtOAc; HPLC (eluent, isooctane-EtOAc); Semi-preparative and then analytical normal-phase HPLC with RI-monitoring.	1D and 2D NMR	1 (0.71)	November/Oualidia, Morocco	[23]
					39 (0.12)
	3.15				9 (0.14)	December/Quiberon, Brittany, France
					15 (0.47)
					21 (0.01)
					22 (0.01)
					25 (0.02)
Et2O S–L extraction, at RT	2.94	Shade-dried and ground	CC (on Silica gel) eluted with a solvent gradient from isooctane to EtOAc Semi-preparative normal phase HPLC (eluent, EtOAc-isooctane), with RI monitoring	HRMS, (1H and 13C) NMR	1 (0.71)	December/Oualidia, Morocco	[46]
					2 (0.03)
					3 (0.08)
					4 (0.01)
					5 (0.004)
					39 (0.12)
					42 (0.01)
Et2O S–L extraction	N.D.	Freeze-dried	Preparative HPLC (DCM-EtOAc, 90:10)	UV, IR, (1H and 13C) NMR, MS	9 (0.28)	-/Loire-Atlantique, France	[19]
					16 (0.06)
Et2O S–L extraction	N.D.	Shade-dried	Separation in different fractions		9	-/Brittany, France	[26]
					15
Et2O S–L extraction, at RT, for 48h	2.2-2.9	Air-dried and ground	CC (on Silica gel) eluted with a solvent gradient from isooctane to EtOAc; Normal-phase HPLC (eluent, isooctane-EtOAc, 2:3), with RI-monitoring	IR, UV, HRMS, 1D and 2D NMR	9 (1.12)	July to June/Roscoff, Brittany, France	[36]
					10 (0.37)
					15 (0.06)
					39 (0.02)
Et2O S–L extraction, at RT	2.40	Freeze-dried and ground	Partitioning between H2O and Et2O; Et2O-soluble material: CC (on Silica gel) eluted with hexane-EtOAc (1:1); HPLC (EtOAc–isooctane), with RI monitoring.	HRMS, EIMS, IR, (1H and 13C) NMR	9 (0.15)	July to August/Roscoff, Brittany, France	[29]
					10 (0.23)
					11 (0.03)
Et2O S–L extraction, at RT	3.15	Shade-dried and ground	CC (on Silica gel) eluted with a solvent gradient from isooctane to EtOAc; Semi-preparative normal-phase HPLC (eluent, EtOAc-isooctane), with RI monitoring	HRMS, EIMS, IR, (1H and 13C) NMR.	9 (0.06)	December/Quiberon and Roscoff, Brittany, France	[31]
					15 (0.01)
					21 (0.01)
					22 (0.02)
					25 (0.03)
	2.40		CC (on Silica gel) eluted with a solvent gradient from hexane to Et2O; HPLC (EtOAc- isooctane, 2:3), with RI monitoring.		9 (0.04)
					10 (0.06)
					11 (0.01)
					12 (0.007)
					15 (0.003)
Et2O S–L extraction	N.D.	Dried	LC using Silica gel; HPLC of the most polar fraction;	IR, MS, NMR	9	-/Quiberon, Brittany, France	[33]
					25
					47
Et2O S–L extraction, at RT	N.D.	Crushed and freeze-dried	Insoluble impurities and pigments elimination with isooctane and EtOH-H2O, respectively; TLC (eluted with Et2O-petroleum ether, 50:50 or DCM-EtOAc 70:30); CC (on Silica gel, eluted with Et2O-petroleum ether, 75:25) Semi-preparative HPLC	UV, IR, (1H and 13C) NMR, MS	15 (2)	-/Loire-Atlantique, France	[20]
Et2O S–L extraction (3x) (3h at 20ºC)	3.67	Freeze-dried and powder	CC (on Silica gel) eluted with a solvent gradient from hexane to Et2O More polar fr.: HPLC with DCM-EtOAc (90:10)	IR, UV, MS, (1H and 13C) NMR	15	January to December/Piriac, France	[27]
					17 (0.01)
Et2O S–L extraction	N.D.	Dried	LC using Silica gel; HPLC of the less polar fr.; Semi-preparative normal-phase HPLC (EtOAc-isooctane, 1:1).	IR, MS, NMR	23 (0.12)	-/El Jadida, Morocco	[32]
					24 (0.08)
					39 (0.01)
DCM S–L extraction, at RT; MeOH S–L extraction	9.06	Freeze-dried	Modified Kupchan method: partitioning between (90:10) MeOH-H2O and n-hexane; MeOH-H2O phase: partitioning between (65:35) MeOH-H2O and CHCl3; Fractionation by flash chromatography (on Silica gel), with EtOAc-n-hexane (80:20) DAD and ELSD-HPLC (using a reverse-phase column), eluent gradient (55:45) MeCN-H2O for 13 min, increasing to 100% MeCN in 5 min, maintaining for 20 min (rt = 16.3 min)	IR, 1D and 2D NMR, HRMS, VCD	14 (0.007)	May/Kilkee, County Clare of Ireland	[50]
MeOH S–L extraction DCM S–L extraction	0.95	Freeze-dried	VLC (on Silica gel) eluted with cyclohexane-EtOAc (1:2); Semi-preparative reverse phase HPLC (gradient of H2O-MeCN) PTLC over Silica gel (n-hexane-EtOAc, 7:3)	(1H, 13C, APT, COSY, HMBC and HSQC) NMR	15 (0.002)	May to June/Peniche, Portugal	[51]
					23 (9.5 × 10−5)
DCM Soxhlet extraction (9 h)	3.92	Freeze-dried and ground		GC-MS	37 (0.06)	May/Ria de Aveiro, Portugal	[38]
					38 (0.10)
					39 (0.01)
					42 (0.01)
					45 (0.003)
					46 (0.01)

^a—% (w/w); ^b—when available, % of dw; ¹H—proton; ¹³C—carbon; 1D and 2D—one and two dimensional; APT—attached proton test; CC—column chromatography; COSY—correlated spectroscopy; DAD—diode array detector; DCM—dichloromethane; EIMS—electron ionization mass spectrometry; ELSD—evaporative light scattering detector; ESI—electrospray ionization; Et₂O—diethyl ether; EtOAc—ethyl acetate; EtOH—ethanol; EY—extraction yield; ext.—extract; fr.—fraction; HMBC—heteronuclear multiple bond correlation; HPLC—high performance liquid chromatography; HRMS—high resolution mass spectrometry; HSQC—heteronuclear single quantum coherence; IR—infrared spectroscopy; LC—liquid chromatography; L–L—liquid–liquid; MeCN—acetonitrile; MeOH—methanol; MS—mass spectrometry; N.D.—nondeterminate; NMR—nuclear magnetic resonance; PTLC—plate thin layer chromatography; Ref—reference; RI—refractive index detector; rt—retention time; RP—reverse phase; RT—room temperature; S–L—solid–liquid; SPE—solid phase extraction; TLC—thin-layer chromatography; UV—ultraviolet spectroscopy; VCD—vibrational circular dichroism; VLC—Vacuum liquid chromatography.