View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Repositorio da Producao Cientifica e Intelectual da Unicamp Hindawi Publishing Corporation International Journal of Analytical Chemistry Volume 2016, Article ID 8361375, 8 pages http://dx.doi.org/10.1155/2016/8361375 Research Article Quantification of Sterol and Triterpenol Biomarkers in Sediments of the Cananéia-Iguape Estuarine-Lagoonal System (Brazil) by UHPLC-MS/MS Giovana Anceski Bataglion,1,2 Hector Henrique Ferreira Koolen,3 Rolf Roland Weber,4 and Marcos Nogueira Eberlin2 1 Department of Chemistry, Federal University of Amazonas (UFAM), 69077-000 Manaus, AM, Brazil 2ThoMSon Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas (UNICAMP), 13083-970 Campinas, SP, Brazil 3DeMpSter Mass Spectrometry Group, Amazonas State University (UEA), 69050-010 Manaus, AM, Brazil 4Marine Organic Chemistry Laboratory, Oceanography Institute, University of Sao˜ Paulo (USP), 05508-120 Sao˜ Paulo, SP, Brazil Correspondence should be addressed to Giovana Anceski Bataglion; [email protected] and Hector Henrique Ferreira Koolen; [email protected] Received 31 December 2015; Accepted 6 March 2016 Academic Editor: Josep Esteve-Romero Copyright © 2016 Giovana Anceski Bataglion et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Sterols and triterpenols present in sedimentary cores from 12 stations along the Cananeia-Iguape´ estuarine-lagoonal system were investigated by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Ten sterols and three triterpenols were identified and quantified, indicating both natural and anthropogenic sources. The relative distributions of sterol and triterpenol showed that the study area is submitted to organic matter (OM) from the Ribeira de Iguape River, seawater, surrounding vegetation, and plankton production. The contribution of these sources depends on the region of the estuarine- lagoonal system and the depth of sediment. Regarding anthropogenic sources, only the samples submitted to freshwater flow from the Ribeira de Iguape River presented concentration of coprostanol higher than the threshold value and diagnostic ratios, coprostanol/(coprostanol + cholestanol) and coprostanol/cholesterol, that indicate moderate contamination by domestic sewage in that area of the estuarine-lagoonal system. Therefore, the approach used herein identified the OM sources and its transport along the Cananeia-Iguape´ estuarine-lagoonal system (Brazil), which is a complex of lagoonal channels located in a United Nations Educational, Scientific and Cultural Organization (UNESCO) Biosphere Reserve. 1. Introduction heterotrophic eukaryote and benthic algae [6]. Furthermore, besides natural OM sources, sterols have been applied in Sterols, also known as alcohol steroids, are hydrophobic environmental studies to track human activities in coastal compounds that occur naturally in most of the living forms areas [7, 8]. [1]. This class of compounds can be easily incorporated into Phytosterols, sterols C28 and C29,comprisethemajor sediments, where they remain well preserved, representing sterol type found in estuarine sediments [9]. Campesterol, good tracers of sources, changes, and preservation of organic -sitosterol, stigmasterol, and stigmastanol are plant sterols; matter (OM) [2]. Sterols have been used to assess the OM thus they are used as geochemical biomarkers to assess the origin in several estuarine systems around the world because plant-derived OM input to aquatic systems [9, 10]. Algae often of their recalcitrance and specific-source, which allows display predominance of C27 sterols (mainly cholesterol), classifying the OM input from different sources, including with compositions different from those observed for vascular terrestrial plants [3], phytoplankton [4], bacteria [5], and plants [11]. In addition, brassicasterol is frequently found in 2 International Journal of Analytical Chemistry several algal classes, whereas cholestanol often occurs in the dinoflagellates class [11]. Other sterols, such as dinosterol and Ribeira de Iguape River ergosterol, are exclusive biomarkers of OM from diatoms and S4 fungi, respectively [12]. Icapara outfall There are also important sterol biomarkers for anthro- S3 pogenicOM,whichareknownasfecalsterols(e.g., S2 Atlantic Ocean S1 coprostanol and epicoprostanol) and indicate contamination N S5 from domestic sewage [13]. Some authors point that absolute S7 S6 W E concentration of coprostanol should not be used as a unique S and conclusive parameter to assess sewage contamination S8 0 510 20 because of the possibility of in situ production. To solve this S9 S10 (km) issueandtoevaluatethedegreeofcontaminationbydomestic Cananéia outfall sewage input, some ratios, such as coprostanol/(coprostanol S11 S12 +cholestanol)andcoprostanol/cholesterol,havebeenpro- posed [14]. Higher plants, especially angiosperms, display triterpenols as typical and exclusive biomarkers. Triterpenols, Figure 1: Map of study area showing the 12 sampling stations such as lupeol and -and -amyrin, are the most common along the Cananeia-Iguape´ estuarine-lagoonal system, located in triterpenols found in sediments of riverine and estuarine Sao˜ Paulo state (Southeast Brazil). Content may not reflect National environments [15]. Geographic’s current map policy. Sources: National Geographic, Estuarine systems are essential compartments involved Esri, DeLorme, HERE, UNEP-WCMC, USGS, NASA, ESA, METI, in the OM recycling and global carbon cycle [16]. Under- NRCAN, GEBCO, NOAA, and Increment P Corp. standing the flows of OM from marine and rivers to estuaries provides information about its cycle in both water column and sediments [17]. Once OM in these systems constitutes a complex mixture of compounds from different sources, a centurycausedanenormousimpactinthisenvironment. reliable technique is required to overcome this complexity After its opening, the channel eroded laterally reaching a [18]. Sterols in sediment samples are generally analyzed by width and depth of approximately 250 and 7m, respectively. gas chromatography-mass spectrometry (GC-MS), which Thus, the channel was renamed to Valo Grande (“Big Scour”) requires a cleanup step using alumina and silica columns, in reference to the dimensions that it acquired. As a result of where sterols are separated from other classes of compounds. this erosion, environmental changes in the Cananeia-Iguape´ The fraction containing sterols is usually derivatized to estuarine-lagoonal system were intensified, since about 60% increase their volatility and decrease the limits of detec- ofthemainflowoftheRibeiradeIguapeRiverwastrans- tion (LOD) [19]. In this sense, liquid chromatography (LC) ferred to the system [22]. The affected area is an important as a separation technique and tandem mass spectrometry ecosystem from the World Heritage List located between (MS/MS) as a detection method are combined to over- two cities (Cananeia´ and Iguape) with a protected green come the inherent challenges of OM complexity [15]. Some area consisting of typical vegetation of salt marshes, Atlantic methods employing analytical platforms based on liquid forest, and mangroves [22]. chromatograph tandem mass spectrometry (LC-MS/MS) for A total of 60 samples from 12 collection sites (0–10 cm the analysis of selected sterols have been described over the depth) were collected at the Cananeia-Iguape´ estuarine- last years for a range of matrices [20, 21] and more recently lagoonal system. Sampling sites with their geographical for sediments [15]. locations and lithological characteristics are shown in Table 1. Sedimentary core samples were collected using a Rossfelder Information on sources, transport, and deposition of OM ∘ is scarce for the Cananeia-Iguape´ estuarine-lagoonal system, VT-1vibracorerandstoredat4C during transportation to which is an important Brazilian ecosystem from the World the laboratory. Prior to the laboratory work, all sediment samples were freeze-dried, pulverized in a mortar, and sieved Heritage List under natural criteria (UNESCO) due to exten- < ∘ sive areas of mangrove and Atlantic forest. Consequently, with a stainless steel sieve ( 250 m)andstoredat4 Cbefore this study aimed to determine the sterol and triterpenol analysis. composition and assess the OM origin in sedimentary cores along the study area. Based on that, an initial insight into 2.2. Chemicals and Reagents. (3)-Cholest-5-en-3-ol (cho- thegeochemistryandflowofOMinthisestuarine-lagoonal lesterol, ≥99% purity), (3,5)-cholestan-3-ol (cholestanol, environment was achieved. >95% purity), (3,5)-cholestan-3-ol (coprostanol, >95% purity), (3,5)-cholestan-3-ol (epicoprostanol, >95% pu- 2. Experimental rity), (3,22E)-ergosta-5,7,22-trien-3-ol (ergosterol, >95% purity), (3,22E)-ergosta-5,22-dien-3-ol (brassicasterol, 2.1. Study Area and Sampling. The Cananeia-Iguape´ estua- >95% purity), (3,24R)-ergost-5-en-3-ol (campesterol, >65% rine-lagoonal system (Figure 1) is located in the Southeast purity), (3)-stigmast-5-en-3-ol (-sitosterol, >98% purity), Region of Brazil (Sao˜ Paulo state). The opening of a 4 km (3,22E)-stigmasta-5,22-dien-3-ol (stigmasterol, >95% puri- long artificial channel linking the Ribeira de Iguape River to ty), (3,5)-stigmastan-3-ol (stigmastanol, >95% purity), the Cananeia-Iguape´