HPLC Pigment Analizinde Kromatografik Ayırma Parametreleri
Yıl 2020,
Cilt: 5 Sayı: 4, 570 - 580, 31.12.2020
Ertuğrul Ağırbaş
,
Osman Akyüz
Öz
Güney Doğu Karadeniz (Artvin-Giresun) kıyıları boyunca Kasım 2014-Ağustos 2015 tarihleri arasında mevsimsel olarak yürütülen bu çalışmada pigment analizinde kullanılan kromatorgrafik ayırma parametreleri ve pigmente dayalı fitoplankton boy gruplarının zamansal ve alansal değişimleri araştırılmıştır. Çalışma kapsamında 12 farklı istasyonda yüzeyden mevsimsel örneklemeler gerçekleştirilmiştir. HPLC-DAD koşullarına bağlı olarak gerçekleştirilen analizler sonucunda 16 standart bileşiğin 38 dakika içerisinde uygun ayırımı elde edilmiştir. Ayırma parametreleri ve çözünürlük değerleri piklerin ayırımının %100’e yakın olduğunu göstermiştir. Kullanılan yöntemin pigment analizi için geçereli olduğu görülmüştür. Çalışma bölgesinde mikrofitoplankton (%8-88) öne çıkan grup olurken bunu pikofitoplankton (%3-60) ve nanofitoplankton (%1-60) takip etmiştir.
Destekleyen Kurum
TÜBİTAK
Teşekkür
Bu çalışma TÜBİTAK 113Y189 numaralı proje ile desteklenmiştir. Deniz çalışmaları esnasında yardımlarından dolayı proje ekibine, “R/V SÜRAT ARAŞTIRMA I” ve “R/V KARADENİZ ARAŞTIRMA” gemilerinin kaptan ve gemi adamlarına teşekkür ederiz.
Kaynakça
- Ağırbaş, E., Koca L. & Aytan U. (2017). Spatio-Temporal Pattern of Phytoplankton and Pigment Composition in Surface Waters of South-Eastern Black Sea. Oceanologia, 59, 283-299.
- Barlow, R.G., Aiken, J., Holligan, P.M., Cummings, D.G., Maritorena, S. & Hooker, S. (2002). Phytoplankton pigment and absorption characteristics along meridional transects in the Atlantic Ocean. Deep-Sea Research Part I-Oceanographic Research Papers, 49 (4), 637-660.
- Barlow, R.G., Aiken, J., Moore, G.F., Holligan, P.M. & Lavender, S. (2004). Pigment adaptations in surface phytoplankton along the eastern boundary of the Atlantic Ocean. Marine Ecology Progress Series, 281, 13-26.
- Brewin, R.J.W., Sathyendranath, S., Hirata, T., Lavender, S.J., Barciela, R.M. & Hardman-Mountford N.J. (2010). A three-component model of phytoplankton size class for the Atlantic Ocean. Ecological Modelling, 221 (11), 1472-1483.
- Booth, B.C., 1993. Estimating cell concentration and biomass of autotrophic plankton using microscopy. In: Kemp P.F., Sherr B.F., Cole J.J. (Eds.), Handbook of methods in aquatic microbial ecology. Lewis Publishers, Boca Raton, FL, pp. 199-205.
- Boyce, D.G., Lewis, M.R. & Worm, B. (2010). Global phytoplankton decline over the past century. Nature, 466 (7306), 591-596.
- Ediger, D., Soydemir, N. & Kideys, A.E. (2006). Estimation of phytoplankton biomass using HPLC pigment analysis in the Southwestern Black Sea. Deep-Sea Research Part II-Topical Studies in Oceanography, 53, 1911-1922.
- Eker-Develi E., Berthon J.F., Canuti, E., Slabakova, N., Moncheva, S., Shtereva G. & Dzhurova, B. (2012). Phytoplankton taxonomy based on CHEMTAX and microscopy in the Northwestern Black Sea. Journal of Marine Systems, 94,18-32.
- Eker-Develi, E., Berthon, J.F. & Linde, D. (2008). Phytoplankton class determination by microscopic and HPLC-CHEMTAX analyses in the southern Baltic Sea. Marine Ecology Progress Series, 359, 69-87.
- Falkowski, P.G. & Raven, J.A. (2007). Aquatic photosynthesis, Princeton University Press, Second edition, 484 s.
- Genç N. (2018). Güneydogu Karadeniz'de pigmente dayalı fitoplankton boy gruplarının fotik bölge dinamiği, (Yüksek Lisans Tezi) Recep Tayyip Erdoğan Üniversitesi, Fen Bilimleri Enstitüsü-, Rize, Türkiye, 48 s.
- Gibb, S.W., Barlow, R.G., Cummings, D.G., Rees, N.W., Trees, C.C., Holligan, P. & Suggett, D. (2000). Surface phytoplankton pigment distributions in the Atlantic Ocean: an assessment of basin scale variability between 50°N and 50°S. Progress in Oceanography, 45, 339-368.
- Gibb, S.W., Cummings, D.G., Irigoien, X., Barlow, R.G., Fauzi, R. & Mantoura C. (2001). Phytoplankton pigment chemotaxonomy of the northeastern Atlantic. Deep Sea Research II, 48, 795-823.
- Gieskes, W.W.C. & Kraay, G.W. (1983). Dominance of Cryptophyceae during the Phytoplankton Spring Bloom in the Central North Sea Detected by HPLC Analysis of pigments. Marine Biology, 75, 179-185.
- Guillard, R.R.L., Murphy, L.S., Foss, P. & Liaaen-Jensen, S. (1985). Synechococcus spp. as likely Zeaksantin-dominant Ultraphytoplankton in the North Atlantic. Limnology and Oceanography, 30, 412-414.
- Jeffrey, S.W. & Vesk, M., 1997. Introduction to marine phytoplankton and their pigment signatures. In: Jeffrey SW, Mantoura RFC, Wright SW (Eds.), Phytoplankton Pigments in Oceanography: Guidelines to Modern Methods. UNESCO, Paris, pp. 19-36.
- Karadeniz, M.N. (2010). Güneydoğu Karadeniz Fitoplankton Pigment Profili Ve Fitoplankton Boy Grupları, Yüksek Lisans Tezi, Recep Tayyip Erdoğan Üniversitesi, Fen Bilimleri Enstitüsü, 70 s.
- Koca, L. (2014). Güney Doğu Karadeniz Kıyıları (Rize) Diatom/Dinoflagellat Oranları Ve Pigment Kompozisyonun Zamansal Değişimi, Yüksek Lisans Tezi, Recep Tayyip Erdoğan Üniversitesi, Fen Bilimleri Enstitüsü, 73 s.
- Kopuz, U., Feyzioglu, A.M. &Agirbas, E. (2012). Picoplankton Dynamics during Late Spring 2010 in the South-Eastern Black Sea. Turkish Journal of Fisheries and Aquatic Sciences, 12, 397-405.
- Mackas, D.L., 2011. Does blending of chlorophyll data bias temporal trend? Nature, 472 (7342), E4-E5.
- Mackey, M.D., Mackey, D.J., Higgins, H.W. & Wright, S.W. (1996). CHEMTAX-a program for estimating class abundances from chemical markers: application to HPLC measurements of phytoplankton. Marine Ecology Progress Series, 144, 265-283.
- Mantoura, R.F.C. & Llewellyn C.A. (1983). The rapid determination of algal Chlorophyll-a and Carotenoid Pigments and Their Breakdown Products in Natural Waters by Reverse-Phase High Performance Liquid Chromatography. Analytica Chimica Acta, 151, 297-314.
- Millie, D.F., Paerl, H.W., Hurley, J.P. & Kirkpatrick, G.J. (1993). Algal pigment determinations in aquatic ecosystems: Analytical Evaluations, applications and recommendations. Current Topics Botincal Research, 1, 1-13. Oceanography UNESCO, Paris, 327-341.
- Obayashi, Y., Tanoue, E., Suzuki, K., Handa, N., Nojiri, Y. & Wong, C.S. (2001). Spatial and Temporal Variabilities of Phytoplankton Community Structure in the Northern North Pacific as Determined by Phytoplankton Pigments. Deep-Sea, Research I, 48, 439-469.
- Skoog, D.A., Holler, F.J. & Nieman, T.A. (2000). Enstrümental Analiz İlkeleri, Birinci Baskı, Saunders Golden Sunburst Publishing, USA, Kılıç E., Köseoğlu F., Yılmaz H., Bilim Yayıncılık, Ankara.
- Stauber, J.L. & Jeffrey, S.W. (1988). Photosynthetic Pigments in fifty-one Species of Marine Diatoms. Journal of Phycology, 24, 158-172.
- Tait, R. V. & Dipper, F. A. (2001). Elements of Marine Ecology, Butterworth-Heinemann Pub., forth edition, Gread Britaiin, 462 s.
- Takahashi, T., Sutherland, S.C., Sweeney, C., Poisson, A., Metzl, N., Tilbrook, B., Bates, N., Wanninkhof, R., Feely, R.A., Sabine, C., Olafsson, J. &
Nojiri, Y. (2002). Global sea-air CO2 flux based on climatological surface ocean pCO2, and seasonal biological and temperature effects. Deep-Sea Research Part II-Topical Studies in Oceanography, 49 (9-10), 1601-1622.
- Türkmen, P. (2016). Güneydoğu Karadeniz Kıyıları (Artvin-Giresun) Pigment Kompozisyonunun Mevsimsel Değişimi, (Yüksek Lisans Tezi) Recep Tayyip Erdoğan Üniversitesi, Rize, Türkiye, 61 s.
- Uitz, J., Claustre, H., Morel, A. & Hooker, S.B. (2006). Vertical distribution of phytoplankton communities in open ocean: An assessment based on surface chlorophyll. Journal of Geophysical Research-Oceans.
- Uitz, J., Claustre, H., Brian Griffiths, F., Ras, J., Garcia, N., & Sandroni, V. (2009). A phytoplankton class-specific primary production model applied to the Kerguelen Islands region (Southern Ocean). Deep-Sea Research I, 56, 541−560.
- Utermohl, H. (1958). Zur Vervollkommnung der quantitativen phytoplankton: Methodik Mitteilung Internationale Vereinigung Theoretische und Angewandte Limnologie 9, 1-38.
- Uysal, Z. (2001). Chroococcoid cyanobacteria Synechococcus spp. in the Black Sea: pigments, size, distribution, growth and diurnal variability. Journal of Plankton Research, 23 (2), 175-189. Uysal, Z. (2006). Vertical distribution of marine cyanobacteria Synechococcus spp. in the Black, Marmara, Aegean and eastern Mediterranean Seas. Deep-Sea Research II, 53, 1976- 1987.
- Wright, S.W. & Jeffrey, S.W. (1987). Fucoxanthin pigment markers of marine phytoplankton analysed by HPLC and HPTLC. Marine Ecology Progress Series, 38, 259-266.
- Wright, S.W. and Thomas, D.P., Marchant, H.J., Higgins, H.W., Mackey, M.D. & Mackey, D.J. (1996). Analysis of Phytoplankton of the Australian Sector of the Southern Ocean: Composition of Microscopy and Size Frequency Data with Interpretations of Pigment HPLC Data Using the CHEMTAX Matrix Factorization Program. Marine Ecology Progress Series, 144, 285-298.
- Zubkov, M.V., Sleigh, M.A., Tarran, G.A., Burkill, P.H. & Leakey, R.J.G. (1998). Picoplanktonic community structure on an Atlantic transect from 50 N to 50 S. Deep Sea Research Part I: Oceanographic Research Papers 45 (8), 1339-1355.
Chromatographic Separation Parameters in HPLC Based Pigment Analysis
Yıl 2020,
Cilt: 5 Sayı: 4, 570 - 580, 31.12.2020
Ertuğrul Ağırbaş
,
Osman Akyüz
Öz
In the present study, chromatographic separation parameters used in pigment analysis and phytoplankton size classes depending on pigments from December 2014 to August 2015 were seasonally investigated along the south-eastern coasts (Giresun-Artvin) of the Black Sea. Along the 12 stations, seasonal samplings were collected from surface waters. Based on the HPLC-DAD conditions, an appropriate separation of 16 pigment standard was achieved within 38 minutes. The separation parameters and the resolution values showed that the peaks were close to 100%. The used method was validated for pigment analysis. Microphytoplankton (8-88%) was the prominent group in the study area, followed by picophytoplankton (3-60%) and nanofitoplankton (1-60%).
Kaynakça
- Ağırbaş, E., Koca L. & Aytan U. (2017). Spatio-Temporal Pattern of Phytoplankton and Pigment Composition in Surface Waters of South-Eastern Black Sea. Oceanologia, 59, 283-299.
- Barlow, R.G., Aiken, J., Holligan, P.M., Cummings, D.G., Maritorena, S. & Hooker, S. (2002). Phytoplankton pigment and absorption characteristics along meridional transects in the Atlantic Ocean. Deep-Sea Research Part I-Oceanographic Research Papers, 49 (4), 637-660.
- Barlow, R.G., Aiken, J., Moore, G.F., Holligan, P.M. & Lavender, S. (2004). Pigment adaptations in surface phytoplankton along the eastern boundary of the Atlantic Ocean. Marine Ecology Progress Series, 281, 13-26.
- Brewin, R.J.W., Sathyendranath, S., Hirata, T., Lavender, S.J., Barciela, R.M. & Hardman-Mountford N.J. (2010). A three-component model of phytoplankton size class for the Atlantic Ocean. Ecological Modelling, 221 (11), 1472-1483.
- Booth, B.C., 1993. Estimating cell concentration and biomass of autotrophic plankton using microscopy. In: Kemp P.F., Sherr B.F., Cole J.J. (Eds.), Handbook of methods in aquatic microbial ecology. Lewis Publishers, Boca Raton, FL, pp. 199-205.
- Boyce, D.G., Lewis, M.R. & Worm, B. (2010). Global phytoplankton decline over the past century. Nature, 466 (7306), 591-596.
- Ediger, D., Soydemir, N. & Kideys, A.E. (2006). Estimation of phytoplankton biomass using HPLC pigment analysis in the Southwestern Black Sea. Deep-Sea Research Part II-Topical Studies in Oceanography, 53, 1911-1922.
- Eker-Develi E., Berthon J.F., Canuti, E., Slabakova, N., Moncheva, S., Shtereva G. & Dzhurova, B. (2012). Phytoplankton taxonomy based on CHEMTAX and microscopy in the Northwestern Black Sea. Journal of Marine Systems, 94,18-32.
- Eker-Develi, E., Berthon, J.F. & Linde, D. (2008). Phytoplankton class determination by microscopic and HPLC-CHEMTAX analyses in the southern Baltic Sea. Marine Ecology Progress Series, 359, 69-87.
- Falkowski, P.G. & Raven, J.A. (2007). Aquatic photosynthesis, Princeton University Press, Second edition, 484 s.
- Genç N. (2018). Güneydogu Karadeniz'de pigmente dayalı fitoplankton boy gruplarının fotik bölge dinamiği, (Yüksek Lisans Tezi) Recep Tayyip Erdoğan Üniversitesi, Fen Bilimleri Enstitüsü-, Rize, Türkiye, 48 s.
- Gibb, S.W., Barlow, R.G., Cummings, D.G., Rees, N.W., Trees, C.C., Holligan, P. & Suggett, D. (2000). Surface phytoplankton pigment distributions in the Atlantic Ocean: an assessment of basin scale variability between 50°N and 50°S. Progress in Oceanography, 45, 339-368.
- Gibb, S.W., Cummings, D.G., Irigoien, X., Barlow, R.G., Fauzi, R. & Mantoura C. (2001). Phytoplankton pigment chemotaxonomy of the northeastern Atlantic. Deep Sea Research II, 48, 795-823.
- Gieskes, W.W.C. & Kraay, G.W. (1983). Dominance of Cryptophyceae during the Phytoplankton Spring Bloom in the Central North Sea Detected by HPLC Analysis of pigments. Marine Biology, 75, 179-185.
- Guillard, R.R.L., Murphy, L.S., Foss, P. & Liaaen-Jensen, S. (1985). Synechococcus spp. as likely Zeaksantin-dominant Ultraphytoplankton in the North Atlantic. Limnology and Oceanography, 30, 412-414.
- Jeffrey, S.W. & Vesk, M., 1997. Introduction to marine phytoplankton and their pigment signatures. In: Jeffrey SW, Mantoura RFC, Wright SW (Eds.), Phytoplankton Pigments in Oceanography: Guidelines to Modern Methods. UNESCO, Paris, pp. 19-36.
- Karadeniz, M.N. (2010). Güneydoğu Karadeniz Fitoplankton Pigment Profili Ve Fitoplankton Boy Grupları, Yüksek Lisans Tezi, Recep Tayyip Erdoğan Üniversitesi, Fen Bilimleri Enstitüsü, 70 s.
- Koca, L. (2014). Güney Doğu Karadeniz Kıyıları (Rize) Diatom/Dinoflagellat Oranları Ve Pigment Kompozisyonun Zamansal Değişimi, Yüksek Lisans Tezi, Recep Tayyip Erdoğan Üniversitesi, Fen Bilimleri Enstitüsü, 73 s.
- Kopuz, U., Feyzioglu, A.M. &Agirbas, E. (2012). Picoplankton Dynamics during Late Spring 2010 in the South-Eastern Black Sea. Turkish Journal of Fisheries and Aquatic Sciences, 12, 397-405.
- Mackas, D.L., 2011. Does blending of chlorophyll data bias temporal trend? Nature, 472 (7342), E4-E5.
- Mackey, M.D., Mackey, D.J., Higgins, H.W. & Wright, S.W. (1996). CHEMTAX-a program for estimating class abundances from chemical markers: application to HPLC measurements of phytoplankton. Marine Ecology Progress Series, 144, 265-283.
- Mantoura, R.F.C. & Llewellyn C.A. (1983). The rapid determination of algal Chlorophyll-a and Carotenoid Pigments and Their Breakdown Products in Natural Waters by Reverse-Phase High Performance Liquid Chromatography. Analytica Chimica Acta, 151, 297-314.
- Millie, D.F., Paerl, H.W., Hurley, J.P. & Kirkpatrick, G.J. (1993). Algal pigment determinations in aquatic ecosystems: Analytical Evaluations, applications and recommendations. Current Topics Botincal Research, 1, 1-13. Oceanography UNESCO, Paris, 327-341.
- Obayashi, Y., Tanoue, E., Suzuki, K., Handa, N., Nojiri, Y. & Wong, C.S. (2001). Spatial and Temporal Variabilities of Phytoplankton Community Structure in the Northern North Pacific as Determined by Phytoplankton Pigments. Deep-Sea, Research I, 48, 439-469.
- Skoog, D.A., Holler, F.J. & Nieman, T.A. (2000). Enstrümental Analiz İlkeleri, Birinci Baskı, Saunders Golden Sunburst Publishing, USA, Kılıç E., Köseoğlu F., Yılmaz H., Bilim Yayıncılık, Ankara.
- Stauber, J.L. & Jeffrey, S.W. (1988). Photosynthetic Pigments in fifty-one Species of Marine Diatoms. Journal of Phycology, 24, 158-172.
- Tait, R. V. & Dipper, F. A. (2001). Elements of Marine Ecology, Butterworth-Heinemann Pub., forth edition, Gread Britaiin, 462 s.
- Takahashi, T., Sutherland, S.C., Sweeney, C., Poisson, A., Metzl, N., Tilbrook, B., Bates, N., Wanninkhof, R., Feely, R.A., Sabine, C., Olafsson, J. &
Nojiri, Y. (2002). Global sea-air CO2 flux based on climatological surface ocean pCO2, and seasonal biological and temperature effects. Deep-Sea Research Part II-Topical Studies in Oceanography, 49 (9-10), 1601-1622.
- Türkmen, P. (2016). Güneydoğu Karadeniz Kıyıları (Artvin-Giresun) Pigment Kompozisyonunun Mevsimsel Değişimi, (Yüksek Lisans Tezi) Recep Tayyip Erdoğan Üniversitesi, Rize, Türkiye, 61 s.
- Uitz, J., Claustre, H., Morel, A. & Hooker, S.B. (2006). Vertical distribution of phytoplankton communities in open ocean: An assessment based on surface chlorophyll. Journal of Geophysical Research-Oceans.
- Uitz, J., Claustre, H., Brian Griffiths, F., Ras, J., Garcia, N., & Sandroni, V. (2009). A phytoplankton class-specific primary production model applied to the Kerguelen Islands region (Southern Ocean). Deep-Sea Research I, 56, 541−560.
- Utermohl, H. (1958). Zur Vervollkommnung der quantitativen phytoplankton: Methodik Mitteilung Internationale Vereinigung Theoretische und Angewandte Limnologie 9, 1-38.
- Uysal, Z. (2001). Chroococcoid cyanobacteria Synechococcus spp. in the Black Sea: pigments, size, distribution, growth and diurnal variability. Journal of Plankton Research, 23 (2), 175-189. Uysal, Z. (2006). Vertical distribution of marine cyanobacteria Synechococcus spp. in the Black, Marmara, Aegean and eastern Mediterranean Seas. Deep-Sea Research II, 53, 1976- 1987.
- Wright, S.W. & Jeffrey, S.W. (1987). Fucoxanthin pigment markers of marine phytoplankton analysed by HPLC and HPTLC. Marine Ecology Progress Series, 38, 259-266.
- Wright, S.W. and Thomas, D.P., Marchant, H.J., Higgins, H.W., Mackey, M.D. & Mackey, D.J. (1996). Analysis of Phytoplankton of the Australian Sector of the Southern Ocean: Composition of Microscopy and Size Frequency Data with Interpretations of Pigment HPLC Data Using the CHEMTAX Matrix Factorization Program. Marine Ecology Progress Series, 144, 285-298.
- Zubkov, M.V., Sleigh, M.A., Tarran, G.A., Burkill, P.H. & Leakey, R.J.G. (1998). Picoplanktonic community structure on an Atlantic transect from 50 N to 50 S. Deep Sea Research Part I: Oceanographic Research Papers 45 (8), 1339-1355.