2
52
Chem. Biodiversity 2016, 13, 249 – 252
Sterol Analysis. The remaining crude extract was fractionated using
a pipette column containing SiO by eluting with 5 ml hexane/AcOEt
[14] E. S. Cho, G. Y. Kim, B. D. Choi, L. L. Rhodes, T. J. Kim, G. H.
Kim, J. D. Lee, Bot. Mar. 2001, 44, 57; J. J. Dorantes-Aranda, L. M.
Garcia-de la Parra, R. Alonso-Rodriguez, L. Morquecho, D.
Voltolina, Environ. Toxicol. 2009, 25, 319.
2
1
2
9 :1 to give the sterol ester fraction, followed by 5 ml hexane/AcOEt
:1 to give the free sterol fraction. Saponification of the two fractions
was accomplished by heating with 10% NaOH/MeOH at 558 for 18 h.
[15] M. P. Mansour, J. K. Volkman, S. I. Blackburn, Phytochemistry
2003, 63, 145.
The mixtures were partitioned between H O and hexane/AcOEt 2 :1;
2
the org. layers were filtered through neutral alumina and concentrated
[16] N. W. Withers, L. J. Goad, T. W. Goodwin, Phytochemistry 1979,
18, 899; W. C. M. C. Kokke, W. Fenical, C. Djerassi, Phytochem-
istry 1981, 20, 127.
to dryness with a stream of N . NMR showed little, if any, sterol in the
2
sterol ester fraction. The free sterol fraction was further fractionated by
reversed-phase HPLC. After evaporation of the HPLC solvent with a
[17] H. R. Harvey, S. A. Bradshaw, S. C. M. Oꢁ Hara, G. Eglinton,
E. D. S. Corner, Phytochemistry 1988, 27, 1723.
1
stream of N , the sterols were characterized by H-NMR. The
2
structures of sterols were assigned by comparison of the NMR data
with authentic standards and literature values. The relative proportions
of the sterols were determined from the integrals of the HPLC
differential refractometer signal.
[18] M. Della Greca, P. Monaco, L. Previtera, Phytochemistry 1989, 28,
629; J.-L. Giner, H. Zhao, D. H. Beach, E. J. Parish, K. Jayasim-
hulu, E. S. Kaneshiro, J. Lipid Res. 2002, 43, 1114; J.-L. Giner, H.
Zhao, Lipids 2004, 39, 763.
[
19] P. Bouvier, M. Rohmer, P. Benveniste, G. Ourisson, Biochem. J.
1976, 159, 267.
J.-L. G. thanks the NSF Biological Oceanography Program for
support (OCE-1061957). Work by C. J. G was supported by New York
Sea Grant Award R/CMB-40.
[20] D. Sica, L. Boniforti, G. Di Giacomo, Comp. Biochem. Physiol. B
1981, 70, 153; W. C. M. C. Kokke, L. Bohlin, W. Fenical, C.
Djerassi, Phytochemistry 1982, 21, 881; A. Kelecom, Bull. Soc.
Chim. Belg. 1981, 90, 971.
[
21] N. W. Withers, W. C. M. C. Kokke, W. Fenical, C. Djerassi, Proc.
Natl. Acad. Sci. U.S.A. 1982, 79, 3764.
REFERENCES
[
[
1] R. M. Kudela, C. J. Gobler, Harmful Algae 2012, 14, 71.
2] Y. Z. Tang, C. J. Gobler, Harmful Algae 2012, 20, 71; R. M. Kudela,
J. P. Ryan, M. D. Blakely, J. Q. Lane, T., D. Peterson, Harmful
Algae 2008, 7, 278; C. J. Gobler, A. Burson, F. Koch, Y. Tang, M. R.
Mulholland, Harmful Algae 2012, 17, 64; Y. Z. Tang, C. J. Gobler,
Mar. Ecol.: Prog. Ser. 2010, 406, 19; X. Jiang, Y. Tang, D. J.
Lonsdale, C. J. Gobler, Mar. Ecol.: Prog. Ser. 2009, 390, 105; Y. Z.
Tang, C. J. Gobler, Mar. Biol. 2009, 156, 2601; Y. Z. Tang, C. J.
Gobler, Harmful Algae 2009, 8, 454.
3] J.-L. Giner, J. A. Faraldos, G. L. Boyer, J. Phycol. 2003, 39, 315.
4] J. K. Volkman, S. M. Barrett, S. I. Blackburn, M. P. Mansour, E. L.
Sikes, F. Gelin, Org. Geochem. 1998, 29, 1163.
5] G. W. Harrington, D. H. Beach, J. E. Dunham, G. G. Holz Jr., J.
Protozool. 1970, 17, 213.
[22] P. D. Nichols, G. J. Jones, J. W. De Leeuw, R. B. Johns, Phytochem-
istry 1984, 23, 1043.
[23] J. K. Volkman, W. I. C. Rijpstra, J. W. De Leeuw, M. P. Mansour,
A. E. Jackson, S. I. Blackburn, Phytochemistry 1999, 52, 659; J. D.
Leblond, P. J. Chapman, J. Phycol. 2002, 38, 670.
[24] J. D. Leblond, A. D. Lasiter, Protist 2012, 163, 38.
[25] M. V. Piretti, G. Pagliuca, L. Boni, R. Pistocchi, M. Diamante, T.
Gazzotti, J. Phycol. 1997, 33, 61.
[26] J. K. Volkman, P. Kearney, S. W. Jeffrey, Org. Geochem. 1990, 15,
489.
[27] N. Withers, in ꢂMarine Natural Products: Chemical and Biological
Perspectivesꢁ, Ed. P. Scheuer, Academic Press, New York, N. Y. ,
1983, Vol. 5, p. 87.
[28] M. Alam, T. B. Sansing, J. R. Guerra, A. D. Harmon, Steroids 1981,
38, 375.
[29] J. Zielinski, W. C. M. C. Kokke, T. B. Tam Ha, A. Y. L. Shu, W. L.
Duax, C. Djerassi, J. Org. Chem. 1983, 48, 3471; G. R. Pettit, D. L.
Herald, C. L. Herald, W. C. M. C. Kokke, C. Djerassi, Steroids
1986, 47, 321.
[30] H. R. Harvey, G. Eglinton, S. C. M. Oꢁ Hara, E. D. S. Corner,
Geochim. Cosmochim. Acta 1987, 51, 3031; S. A. Bradshaw,
S. C. M. Oꢁ Hara, E. D. S. Corner, G. Eglinton, J. Mar. Biol. Assoc.
U. K. 1990, 70, 771; S. A. Bradshaw, S. C. M. Oꢁ Hara, E. D. S.
Corner, G. Eglinton, J. Mar. Biol. Assoc. U. K. 1991, 71, 635.
[31] H. R. Harvey, S. C. M. Oꢁ Hara, G. Eglinton, E. D. S. Corner, Org.
Geochem. 1989, 14, 635; H. M. Talbot, R. N. Head, R. P. Harris,
J. R. Maxwell, Org. Geochem. 2000, 31, 871.
[
[
[
[
[
6] V. I. Svetashev, A. B. Imbs, J. Phycol. 2014, 50, 322.
7] I. Lang, L. Hodac, T. Friedl, I. Feussner, BMC Plant Biol. 2011, 11,
1
24.
[
[
8] M. V. Bell, J. R. Dick, D. W. Pond, Phytochemistry 1997, 45, 303; J.-
A. Marshall, P. D. Nichols, G. M. Hallegraeff, J. Appl. Phycol.
2
002, 14, 255; G. E. Napolitano, W. M. N. Ratnayake, R. G.
Ackman, Phytochemistry 1988, 27, 1751; J.-L. Giner, H. Zhao, C.
Tomas, Phytochemistry 2008, 69, 2167.
9] P. G. Thomson, S. W. Wright, C. J. S. Bolch, P. D. Nichols, J. H.
Skerratt, A. McMinn, J. Phycol. 2004, 40, 867.
[
10] M. P. Mansour, J. K. Volkman, A. E. Jackson, S. I. Blackburn, J.
Phycol. 1999, 35, 710.
[
[
11] J. D. Leblond, P. J. Chapman, J. Phycol. 2000, 36, 1103.
12] C. C. Parrish, G. Bodennec, P. Gentien, A. S. W. deFreitas,
ꢂProduction of Signature and Potentially Toxic Fatty Acids by
Gymnodinium c.f. nagasakiense and Pseudo-nitzschia multiseriesꢁ,
in ꢂCanadian Technical Report of Fisheries and Aquatic Sciences
No. 2138ꢁ, Ed. R. W. Penney, Dept. of Fisheries and Oceans
Science Branch, St. Johnꢁs, NF, Can., 1996, p. 15; F. Sellem, D.
Pesando, G. Bodennec, A. El Abed, J.-P. Girard, Water Res. 2000,
[32] J.-L. Giner, H. Zhao, M. S. Dixon, G. H. Wikfors, Comp. Biochem.
Physiol., Part B: Biochem. Mol. Biol. 2016, 192, 1.
[33] C. Gobler, D. Berry, O. Anderson, A. Burson, F. Koch, B. Rodgers,
L. Moore, J. Goleski, B. Allam, P. Bowser, Y. Tang, R. Nuzzi,
Harmful Algae 2008, 7, 293.
[34] M. Doblin, S. Blackburn, G. Hallegraeff, J. Exp. Mar. Biol. Ecol.
1999, 236, 33.
3
4, 550.
[
13] C. Ghioni, A. E. A. Porter, I. H. Sadler, D. R. Tocher, J. R.
Received July 9, 2015
Sargent, Lipids 2001, 36, 145.
Accepted August 14, 2015
www.cb.wiley.com
ꢀ 2016 Verlag Helvetica Chimica Acta AG, Zürich