Fitoplankton Sınıflarının Belirlenmesinde HPLC ve Spektroflorometrik Yöntemlerin Karşılaştırılması

Öz

Bu çalışmada fitoplankton gruplarının tespit edilmesinde yaygın olarak kullanılan Yüksek Performanslı Sıvı Kromatografik (HPLC) ve spektroflorometrik metotlar kıyaslanmıştır. Bu amaçla Mersin Körfezi – Rodos Adası arasında farklı ekosistemleri temsilen 5 farklı istasyondan 4 farklı dönemde (Temmuz-Eylül 2012 ve Mart-Mayıs 2013) 152 adet su örneği alınarak farklı tip filtrelere süzülmüştür. Klorofil-a, pikoplankton, nanoplankton ve mikroplankton parametreleri her iki metotla ölçülerek kıyaslanmıştır. HPLC cihazının spektroflorometreye göre daha hassas olduğu fakat yöntemde kullanılan GF/F filtrelerin pikoplanktonik hücreleri tutmada başarısız olduğu belirlenmiştir. Akdeniz gibi pikoplankton yoğunluğunun fazla olduğu denizlerde kullanılan GF/F filtreler, nükleopor polikarbonat filtrelere göre biyokütle kaybına neden olarak yanlış okumalara neden olabilmektedir. Toplam klorofil-a ve mikroplankton ölçümlerinde her iki yöntem birbirleriyle uyumlu bulunmuştur. 

Anahtar Kelimeler

• Klorofil-a,Pigment,Pikoplankton,Nanoplankton,Mikoplankton

Comparison of HPLC and Spectrofluorometric Method for Determination of Phytoplankton Classess

Abstract

In this study, High Performance Liquid Chromatographic (HPLC) and Spectrofluorometric methods commonly used in determining of phytoplankton groups were compared. For this purpose, totally 152 water samples were collected in four different periods (July-September 2012 and March-May 2013) from five stations representing different ecosystems between Mersin Bay and Rhodes Island filtered by using different types of filters. Chlorophyll a, picoplankton, nanoplankton and microplankton parameters were compared by using HPLC and spectrofluorometric methods. The HPLC was found to be more sensitive than the spectrofluorometer, but it was determined that the GF/F filters used in the method failed to retain the picoplanktonic cells. GF/F filters used in seas where the concentration of picoplankton is high, such as the Mediterranean, may cause biomass loss as compared to nucleopore polycarbonate filters, causing false readings. Chlorophyll-a and microplankton measurements were found to be compatible with each other when using both methods.

Keywords

• Cholorophyll-a,Pigments,Picoplankton,Nanoplankton,Microplankton

References

1. Ağırbaş, E. 2016. Fitoplankton Pigment Konsantrasyonunun ve Boy Gruplarının HPLC Tekniği ile Belirlenmesi (Güneydoğu Karadeniz, Rize). Yunus Araştırma Bülteni, (2): 81-90, doi:10.17693/yunus.20918
2. Barlow R. G., Mantoura R. F. C., Gough M. A. ve Fileman T. W. 1993. Pigment signatures of the phytoplankton composition in the north-eastern Atlantic during the 1990 spring bloom. Deep Sea Research, II, 40 (1-2): 459-477. doi:10.1016/0967-0645(93)90027-K.
3. Bidigare, R. R., Ondrusek, M. E., Morrow, J. H. ve Kiefer, D. A. 1990. In vivo adsorption properties of algal pigments. SPIE Ocean Optics, 1302: 290–302. doi:10.1117/12.21451.
4. Burford, M. A. ve Pollard, P. C. 1994. Pigment contaminants in polycarbonate filters. Marine Ecology Progress Series, 103: 203-206
5. Gibb, S. W., Barlow, R. G. ve Cummings, D. G. 2000. Surface phytoplankton pigment distributions in the Atlantic Ocean: anassessment of basin scale variability between 50° N and 50° S. Progress in Oceanography, 45: 339–368, doi:10.1016/S0079-6611(00)00007-0
6. Gibb, S. W., Cummings, D. G., Irigoien, X., Barlow, R. G., Fauzi, R. ve Mantoura, C. 2001. Phytoplankton pigment chemotaxonomy of the northeastern Atlantic. Deep Sea Research. Part II: Topical Studies in Oceanography, 48 (4-5): 795–823. doi:10.1016/s0967-0645(00)00098-9
7. Jeffrey, S. W., Mantoura, R. F. C. ve Wright, S. W. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing Paris, 37–84, ISBN: 9231032755.
8. Laxen, D. P. H. ve Chandler, I. M. 1982. Comparison of filtration techniques for size distribution in freshwater Anal. Chem., 54, pp. 1350–1355

9. Li, W. K. W., Zohary, T., Yacobi, Y. Z. ve Wood, A. M. 1993. Ultraphytoplankton in the eastern Mediterranean Sea: towards deriving phytoplankton biomass from flow cytometric measurements of abundance, fluorescence and light scatter. Marine Ecology Progress Series, 102: 79-87.
10. Mantoura, R. F. C. ve Llewellyn, C. A. 1983. The rapid determination of algal chlorophyll and carotenoid pigments and their breakdown products in natural waters by reverse-phase high-performance liquid chromatography. Analytica Chimica Acta, 151:297-314. doi:10.1016/S0003-2670(00)80092-6.
11. Murray, A. P., Gibbs, C. F., Longmore, A. R. ve Flett, D. J. 1986. Determination of chlorophyll in marine waters: intercomparison of a rapid HPLC method with full HPLC, spectrophotometric and fluorimetric methods. Marine Chemistry, 19:211-227
12. Olenina, I., Hajdu, S., Edler, L., Andersson, A., Wasmund, N., Busch, S., Gobel, J., Gromisz, S., Huseby, S., Huttunen, M., Jaanus, A., Kokkonen, P., Ledaine, I., ve Niemkiewicz, E. 2006. Biovolumes and size-classes of phytoplankton in the Baltic Sea, HEL-COM Baltic Sea Environment Protection Commission, 106, 1–144, 2006.
13. Strickland, J. D. H. ve Parsons, T. R. 1972. A Practical Handbook of Seawater Analysis, 2nd edition. Bulletin of the Fisheries Research Board of Canada, No. 167, 310 pp.
14. UNEP/MAP. 2005. Sampling and analysis techniques for the eutrophication monitorıng strategy of MED POL, MAP Technical Reports Series No. 163, Athens.
15. Vidussi, F., Claustre, H., Manca, B., Luchetta, A. ve Marty, J. C. 2001. Phytoplankton pigment distribution in relation to upper thermocline circulation in the eastern Mediterranean Sea, during winter. Journal of Geophysical Research, 106, 19, 939–19, 956.
16. Morán X. A. G., Gasol, J. M., Arin, L. ve Estrada, M. 1999. A comparison between glass fiber and membrane filters for the estimation of phytoplankton POC and DOC production. Marine Ecology Progress Series 187:31– 41
17. Yücel, N. 2013. Monthly Changes in Primary and Bacterial Productivity in the North-Eastern Mediterranean Shelf Waters. Doktora Tezi. Mersin, Ortadoğu Teknik Üniversitesi.
18. Yücel, N. ve Uysal, Z. 2017. Kuzeydoğu Akdeniz’de Kıyısal ve Açık sularda Fitoplankton Dinamikleri. Yunus Araştırma Bülteni, 17 (1), doi: 10.17693/yunusae.v17i26557.280523