Comparative metabolic study of the chloroform fraction of three Cystoseira species based on UPLC/ESI/MS analysis and biological activities

Figures & data

Table 1. Metabolite profiling of C. myrica, C. trinodis and C. tamariscifolia Chloroform fraction using UPLC-ESI-MS in the negative ion mode.

Peak No.	Rt. (min)	Compound Name	[M − H] ^− (m/z)	Molecular Formula	Chemical class	Area %			References
						C. myrica	C. trinodis	C. tamariscifolia
1	1.07	(+)-Afzelechin	273.00	C15H14O5	Flavonoid	1.37	1.60	1.73	^74
2	1.36	Resveratrol	227.10	C14H12O3	Stilbene	10.04	22.18	–	^75
3	1.91	(2E, l0E)-1-Hydroxy-6,13-diketo-7- methylene-3,11,15-trimethylhexadeca-2, l0, l4-triene	317.10	C20H30O3	Acyclic diterpene	0.76	2.03	–	^50
4	2.19	6-Methoxy-4-methylcoumarin	189.15	C11H10O3	Coumarin	–	–	3.05	^76
5	2.91	Syringic acid	197.85	C9H10O5	Phenolic acid	1.36	2.27	–	^75
6	7.74	4′,7-dimethoxyluteolin	313.30	C₁₇H₁₄O₆	Flavonoid	–	–	2.63	77
7	8.64	Octadecanoic acid, ethyl ester	311.30	C20H40O2	Fatty acid	–	–	4.58	^78
8	9.42	Cystomexicone B	357.20	C22H30O4	Nor sesquiterpenoid	–	–	0.36	^72
9	10.27	Sagerinic acid	719.45	C36H32O16	Lignan	5.66	3.19	–	^79
10	11.15	Eicosaenoic acid	309.30	C20H37O2	Fatty acid	2.09	1.13	–	^80
11	11.35	Naringin	579.40	C27H32O14	Flavonoid	–	0.80	–	^81
12	11.65	Compound 5	251.10	C16H28O2	Meroditerpenoids	–	–	2.41	^52
13	11.76	Neoeriotrin	595.40	C27H32O15	Flavonoid	–	0.18	–	^81
14	12.14	4′,12-Di-O-Acetylcystone F	557.35	C32H45O8	Meroditerpenoids	3.58	1.06	–	^15
15	12.15	p-Coumaric acid glucoside	325.30	C15H17O8	Phenolic acid	–	–	0.71	^77
16	12.27	Docosanedioic acid	369.20	C22 H42 O4	Fatty acid	–	0.42	–	^82
17	12.51	Catechin dimer-deoxyhexose	723.45	–	Catechin	1.67	3.20	–	^83
18	12.99	Hydroxy-linolenic acid	293.25	C18H30O3	Fatty acid	3.47	0.59	–	^84
19	13.22	1-O-Caffeoyl-3-O-sinapoylquinic acid	559.40	C27H28O13	Phenolic acid	1.79	3.72	–	^85
20	13.44	Quercetin	301.25	C15H10O7	Flavonoid	3.30	–	–	^86
21	13.54	Pachydictyol A	287.30	C20H32O	Diterpene	–	–	29.84	^12
22	13.99	Phytol	295.30	C18H32O3	Diterpene	3.45	–	–	^51
23	14.17	3[A]chlorobifuhalol hexacetate	535.40	C24H21ClO12	Phlorotannins	–	2.47	–	^57
24	14.45	(2E,10E)-1,6-dihydroxy-7-methy1ene-13-keto-3,11, 15-trimethylhexadeca-2,10,14-triene	319.30	C20H32O3	Acyclic diterpene	3.83	2.10	–	^50
25	14.62	Quercetin 7-O-galloyl-hexoside	585.40	C28H24O16	Flavonoid	1.00	0.81	–	^87
26	15.32	Dihydroxy hexadecanoic acid	287.30	C16H31O4	Fatty acid	3.79	–	2.86	^80
27	15.45	2′′,3′′-Dihydro-3′,3′′′-biapigenin methyl ether	553.35	C31H22O10	Flavonoid	2.52	7.49	–	^88
28	15.67	Alternariol 9-methyl ether	271.25	C15H12O5	Dibenzo-α-pyrone analog	8.87	–	–	^89
29	15.89	Salvianolic acid F isomer	313.30	C17H14O6	Stilbene	–	0.98	1.85	^79
30	16.00	Isorhamnetin 7-O-[3-hydroxy-3 -methylglutaroyl]hexoside	621.50	–	Flavonoid	2.90	–	–	^90
31	16.73	Cystophloroketal A	563.45	C34H44O7	Phloroglucinol − meroterpenoid hybrid	2.17	–	–	^54
32	16.89	Lithospermic acid A	537.40	C27H22O12	Benzofuran	–	4.07	1.70	^79
33	17.79	Cystophloroketal B	563.40	C34H44O7	Phloroglucinol − meroterpenoid hybrid	24.72	21.71	1.11	^54
34	17.98	Octadecatrienoic acid	277.20	C18H29O2	Fatty acid	–	–	2.03	^80

Figure 1. Chemical structures of the major characterised metabolites in three Cystoseira species using UPLC-ESI-MS in the negative ion mode.

Table 2. Total phenolic and flavonoid content of C. myrica, C. trinodis and C. tamariscifolia chloroform fraction.

Extracts	Total phenolics (mg/g gallic acid equivalents)	Total flavonoids (mg/g quercetin equivalents)
C. myrica	16.75 ± 1.03	25.41 ± 0.84
C. trinodis	17.51 ± 0.85	35.48 ± 3.14
C. tamariscifolia	13.34 ± 0.62	9.76 ± 1.25

Values are mean ± SEM, n = 3

Table 3. Antioxidant activity (IC₅₀) of C. myrica, C. trinodis and C. tamariscifolia chloroform fraction using DPPH and FRAP assays.

Extracts	IC50 (µg/mL)
	Scavenging ability on DPPH radicles	Scavenging ability in FRAP assay
C. myrica	494.19 ± 14.23	NA
C. trinodis	13.79 ± 1.09	26.32 ± 1.97
C. tamariscifolia	150.07 ± 8.12	284.73 ± 15.21
Ascorbic acid (Standard)	10.22 ± 0.64	20.89 ± 1.25

Values are mean ± SEM, n = 3

IC₅₀: Inhibitory concentration 50%

NA: No activity

Table 4. Anti-hyperglycaemic activity and Anti-inflammatory activity % (IC₅₀) of C. myrica, C. trinodis and C. tamariscifolia chloroform fraction.

Extracts	IC50 (µg/mL)
	α-Amylase	α-Glucosidase	COX-1
C. myrica	68.71 ± 4.09	46.91 ± 3.68	19.76 ± 1.46
C. trinodis	24.81 ± 2.34	20.29 ± 1.87	16.13 ± 0.89
C. tamariscifolia	46.29 ± 2.91 µg	28.51 ± 1.87	52.19 ± 3.12
Acarbose (Standard)	12.29 ± 0.73	3.12 ± 0.28	–
Celecoxib (Standard)	–	–	9.07 ± 0.67

Values are mean ± SEM, n = 3

IC₅₀: Inhibitory concentration 50%

NA: No activity

Table 5. The docking scores achieved by the major identified compounds towards various enzymes.

Compound Name	Docking Score (Kcal/Mol)
	α-Amylase (7TAA)	α-Glucosidase (7KB8)	COX-1 (5WBE)	Tyrosinase (5M8Q)
Resveratrol	−12.51	−11.42	−12.86	−9.59
(2E, l0E)-1-Hydroxy-6,13-diketo-7- methylene-3,11,15-trimethylhexadeca-2, l0, l4-triene	−10.17	−12.75	−12.69	−7-07
6-Methoxy-4-methylcoumarin	−8.23	−8.66	−10.19	−7.62
4′,7-dimethoxyluteolin	−13.69	−13.38	−17.05	−9.83
Eicosanoic acid	−10.11	−10.69	−12.17	−9.15
Naringin	−13.91	−17.24	−20.29	−11.51
Hydroxy-linolenic acid	−10.03	−11.69	−12.97	−9.90
Quercetin	−13.52	−13.32	−15.43	−13.01
Pachydictyol A	−10.22	−12.81	−12.41	−8.52
Phytol	−9.28	−10.00	−12.01	−10.56
(2E,10E)-1,6-dihydroxy-7-methy1ene-13-keto-3,11,15-trimethylhexadeca-2,10,14-triene	−9.40	−10.53	−12.71	−7.81
2′′,3′′-Dihydro-3′,3′′′-biapigenin methyl ether	−14.93	−14.88	−18.07	−14.81
Compound 5	−8.42	−10.71	−10.92	−7.83
4′,12-Di-O-Acetylcystone F	−12.56	−12.09	−16.11	−9.51
Cystophloroketal A	−10.87	−12.91	−16.09	−11.23
Cystophloroketal B	−12.22	−12.58	−7.72	−11.61

Figure 2. (I) 2D binding modes of (A) 4′,7-Dimethoxyluteolin, (B) 2″,3″-Dihydro-3',3'″-biapigenin methyl ether, (C) Naringin to the active binding sites of α-amylase enzyme. (II) 2D binding modes of (A) 4′,7-dimethoxyluteolin, (B) 2″,3″-dihydro-3',3'″-biapigenin methyl ether, (C) Naringin to the active binding sites of α-glucosidase enzyme.

Figure 3. (I) 2D binding modes of (A) 4',7-Dimethoxyluteolin, (B) 2'',3''-Dihydro-3',3'''-biapigenin methyl ether, (C) Naringin to the active binding sites of COX-1 enzyme. (II) 2D binding modes of (A) Cystophloroketal B, (B) 2'',3''-Dihydro-3',3'''-biapigenin methyl ether, (C) Quercetin to the active binding sites of Tyrosinase enzyme.

Enomoto H, Takahashi S, Takeda S, Hatta H. Distribution of flavan-3-ol species in ripe strawberry fruit revealed by matrix-assisted laser desorption/ionization-mass spectrometry imaging. Molecules. 2020;25(1):2373.

 PubMed Web of Science ®Google Scholar

Sun J, Liang F, Bin Y, Li P, Duan C. Screening non-colored phenolics in red wines using liquid chromatography/ultraviolet and mass spectrometry/mass spectrometry libraries. Molecules. 2007;12(3):679–693.

 PubMed Web of Science ®Google Scholar

Amico V, Oriente G, Piattelli M, Ruberto G, Tringali C. Novel acyclic diterpenes from the brown alga Cystoseira crinita. Phytochemistry. 1981;20(5):1085–1088.

 Web of Science ®Google Scholar

Tine Y, Renucci F, Costa J, Wélé A, Paolini J. A method for LC-MS/MS profiling of coumarins in zanthoxylum zanthoxyloides (Lam.) B. Zepernich and timler extracts and essential oils. Molecules. 2017;22(1):174.

 PubMed Web of Science ®Google Scholar

Simirgiotis MJ, Benites J, Areche C, Sepúlveda B. Antioxidant capacities and analysis of phenolic compounds in three endemic nolana species by HPLC-PDA-ESI-MS. Molecules. 2015;20(6):11490–11507.

 PubMed Web of Science ®Google Scholar

Ads EN, Rajendrasozhan S, Hassan SI, Sharawy SMS, Humaidi JR. Phytochemical screening of different organic crude extracts from the stem bark of Ziziphus spina-christi (L.). biomedicalresearch. 2018;29(8):1645–1652.

 Google Scholar

Zbakh H, Zubía E, de los Reyes C, Calderón-Montaño JM, López-Lázaro M, Motilva V. Meroterpenoids from the brown alga cystoseira usneoides as potential anti-inflammatory and lung anticancer agents. Mar Drugs. 2020;18(4):207.

 PubMed Web of Science ®Google Scholar

Barros L, Dueñas M, Dias MI, Sousa MJ, Santos-Buelga C, Ferreira ICFR. Phenolic profiles of cultivated, in vitro cultured and commercial samples of Melissa officinalis L. infusions. Food Chem. 2013;136(1):1–8.

 PubMed Web of Science ®Google Scholar

Farag MA, Gad HA, Heiss AG, Wessjohann LA. Metabolomics driven analysis of six Nigella species seeds via UPLC-qTOF-MS and GC-MS coupled to chemometrics. Food Chem. 2014;151:333–342.

 PubMed Web of Science ®Google Scholar

Li X, Xiao H, Liang X, Shi D, Liu J. LC-MS/MS determination of naringin, hesperidin and neohesperidin in rat serum after orally administrating the decoction of Bulpleurum falcatum L. and Fractus aurantii. J Pharm Biomed Anal. 2004;34(1):159–166.

 PubMed Web of Science ®Google Scholar

Mokrini R, Mesaoud M, Ben Daoudi M, Hellio C, Maréchal JP, El Hattab M, Ortalo-Magné A, Piovetti L, Culioli G. Meroditerpenoids and derivatives from the brown alga Cystoseira baccata and their antifouling properties. J Nat Prod. 2008;71(11):1806–1811.

 PubMed Web of Science ®Google Scholar

De Los Reyes C, Ortega MJ, Zbakh H, Motilva V, Zubía E. Cystoseira usneoides: A Brown Alga Rich in Antioxidant and Anti-inflammatory Meroditerpenoids. J Nat Prod. 2016;79(2):395–405.

 PubMed Web of Science ®Google Scholar

Patil SH, Kurlapkar DD, Gaikwad DK. Phytochemical characterization of natural dye extracted from Senna siamea Pods. Open Access Libr J. 2020;07(04):1–11.

 Google Scholar

de Souza Mesquita LM, Rodrigues CFB, da Rocha CQ, Bianchim MS, Rodrigues CM, de Oliveira VM, Gaeta HH, Belchor MN, Toyama MH, Vilegas W. LC–ESI–IT-MS/MS and MALDI-TOF Approach: identification of natural polymers from Rhizophora mangle barks and determination of their analgesic and anti-inflammatory properties. Nat Prod Bioprospect. 2019;9(1):23–34.

 PubMed Web of Science ®Google Scholar

Ayoub IM, Korinek M, El-Shazly M, Wetterauer B, El-Beshbishy HA, Hwang T-L, Chen B-H, Chang F-R, Wink M, Singab ANB, et al. Activity of Chasmanthe aethiopica leaf extract and its profiling using LC/MS and GLC/MS. Plants. 2021;10(6):1118.

 Google Scholar

Ben SR, Hamed AI, Mahalel UA, Al-Ayed AS, Kowalczyk M, Moldoch J, Oleszek W, Stochmal A. Tentative characterization of polyphenolic compounds in the male flowers of Phoenix dactylifera by liquid chromatography coupled with mass spectrometry and DFT. Int J Mol Sci. 2017;18(3):1–18.

 Web of Science ®Google Scholar

Chen G, Li X, Saleri F, Guo M. Analysis of flavonoids in Rhamnus davurica and its antiproliferative activities. Molecules. 2016;21(10):1275.

 PubMed Web of Science ®Google Scholar

Ayyad SEN, Abdel-Halim OB, Shier WT, Hoye TR. Cytotoxic hydroazulene diterpenes from the brown alga Cystoseira myrica. Z Naturforsch C J Biosci. 2003;58(1-2):33–38.

 PubMed Web of Science ®Google Scholar

El Amrani Zerrifi S, El Khalloufi F, Mugani R, El Mahdi R, Kasrati A, Soulaimani B, Barros L, Ferreira ICFR, Amaral JS, Finimundy TC, et al. Seaweed essential oils as a new source of bioactive compounds for cyanobacteria growth control: Innovative ecological biocontrol approach. Toxins (Basel). 2020;12(8):527.

 PubMed Web of Science ®Google Scholar

Glombitza K, Schmidt A. Nonhalogenated and halogenated phlorotannins from the brown alga Carpophyllum angustifolium. J Nat Prod. 1999;62(9):1238–1240.

 PubMed Web of Science ®Google Scholar

Salih EYA, Fyhrquist P, Abdalla AMA, Abdelgadir AY, Kanninen M, Sipi M, Luukkanen O, Fahmi MKM, Elamin MH, Ali HA. LC-MS/MS tandem mass spectrometry for analysis of phenolic compounds and pentacyclic triterpenes in antifungal extracts of Terminalia brownii (Fresen). Antibiotics (Basel). 2017;6(4):37.

 PubMed Web of Science ®Google Scholar

Yao H, Chen B, Zhang Y, Ou H, Li Y, Li S, Shi P, Lin X. Analysis of the total biflavonoids extract from Selaginella doederleinii by HPLC-QTOF-MS and its in vitro and in vivo anticancer effects. Molecules. 2017;22(2):325.

 PubMed Web of Science ®Google Scholar

Lou J, Yu R, Wang X, Mao Z, Fu L, Liu Y, Zhou L. Alternariol 9-methyl ether from the endophytic fungus Alternaria sp. Samif01 and its bioactivities. Braz J Microbiol. 2016;47(1):96–101.

 PubMed Web of Science ®Google Scholar

López-Angulo G, Montes-Avila J, Díaz-Camacho SP, Vega-Aviña R, López-Valenzuela JÁ, Delgado-Vargas F. 2018. Comparison of terpene and phenolic profles of three wild species of Echeveria (Crassulaceae). J Appl Bot Food Qual. 91:145–154.

 Google Scholar

El Hattab M, Genta-Jouve G, Bouzidi N, Ortalo-Magné A, Hellio C, Maréchal JP, Piovetti L, Thomas OP, Culioli G. Cystophloroketals A-E, Unusual Phloroglucinol-Meroterpenoid Hybrids from the Brown Alga Cystoseira tamariscifolia. J Nat Prod. 2015;78(7):1663–1670.

 PubMed Web of Science ®Google Scholar