the Creative Commons Attribution 4.0 License.
the Creative Commons Attribution 4.0 License.
Technical note: Measurements of fluorescent dissolved organic matter (FDOM) in seawater (Filter blanks, pore sizes, and storage)
Abstract. Fluorescent dissolved organic matter (FDOM) provides crucial information regarding the sources and characteristics of DOM in oceans. However, the FDOM measurements vary depending on filter blanks, pore sizes, and sample storage. To develop more reliable methods for FDOM measurements, in this study, we examined the uncertainties associated with different preparation methods for seawater samples. These samples identified three primary components using parallel factor analyses: terrestrial humic-like peak (C peak), marine humic-like peak (M peak), and protein-like peak (T peak). Relatively high procedural blank values were obtained from samples filtered through the pre-combusted glass fiber filter (0.7 μm pore size) and membrane filter (0.2 μm pore size) without pre-cleaning. However, the blank values became negligible when the filter was pre-washed with 5 mL of 0.1 M HCl or 20 mL of distilled water. The effects of different filter pore sizes were not observed for the C and M peak concentrations (FDOMH), but relatively low T peak values were observed for filtered samples (0.7 or 0.2 μm) relative to unfiltered samples. For all samples, FDOMH showed consistent results for 21 days (8 % ± 3 %) when stored in a refrigerator or a freezer. However, T peak concentration decreased rapidly in both filtered (15 %–50 %) and unfiltered samples (10 %–40 %) after five days, indicating considerable bacterial degradation of protein-like components within three days. Therefore, our results suggest that reliable FDOMH values can be obtained either unfiltered and filtered samples stored in either a refrigerator or freezer for three weeks, but careful sample filtration, storage, blank controls are necessary for T peak measurements.
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Status: final response (author comments only)
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RC1: 'Comment on egusphere-2025-501', Philippe Massicotte, 27 Feb 2025
The comment was uploaded in the form of a supplement: https://egusphere.copernicus.org/preprints/2025/egusphere-2025-501/egusphere-2025-501-RC1-supplement.pdf
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AC1: 'Reply on RC1', Junhyeong Seo, 02 Apr 2025
The comment was uploaded in the form of a supplement: https://egusphere.copernicus.org/preprints/2025/egusphere-2025-501/egusphere-2025-501-AC1-supplement.pdf
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EC1: 'Reply on AC1', Huixiang Xie, 19 Apr 2025
Dear authors,
Your responses to most of both referees’ comments do not provide any detailed information on how you will address the referees’ concerns and what relevant revisions you will make. For example, in many cases, your responses are simply “revised to clarify” but fail to show what specific revisions you will make. Another example: “More explanations are added”. What are the “more explanations”? please.
I cannot make a decision on the manuscript until you provide detailed responses (including the planned revision texts, if applicable) to each of the referees’ comments. A lack of substance in your responses will also make it difficult for the referees to re-review the manuscript, if needed.
Please note that Referee#2 made many suggestions and comments in the annotated manuscript copy. You should respond to these suggestions and comments as well.
In addition to the referees’ comments, I also have a few comments on the manuscript:
Line 31-32: Siegel et al. (2002) studied CDM (colored dissolved and detrital organic materials) not FDOM (FDOM is only part of CDOM). In addition, it is the absorption of UV by CDOM not FDOM that affects photosynthesis and the growth of marine microorganisms. Do not mess up FDOM with CDOM. These references are not appropriate for the statements that you made for FDOM.
Line 42: “formalin”. I do not think Spencer et al. (2007) used formalin. Please check. If you find it in their paper, please let me know on what page it appears.
Line 46-47: “various sampling and storage methods”. The preceding two statements appear to indicate that there is a generally accepted protocol for FDOM sampling and sample treatment. Reword the preceding statements.
Line 47: “for different DOM compositions”. You were focused on FDOM earlier now suddenly switched to DOM. Be consistent.
Line 86: Coble (1996) and Coble et al. (1998) did not report PARAFAC modeling.
Section 3.2: Add a comparison between the 0.2 um and 0.7 um filtration?
Section 3.3: Add comparisons among different depths and different filter pore sizes? Figures show that different depths and pore sizes gave different results.
Please resubmit your responses before May 9, 2025.
Sincerely,
Huixiang Xie
Associate Editor
Citation: https://doi.org/10.5194/egusphere-2025-501-EC1 -
AC3: 'Reply on AC1', Junhyeong Seo, 30 Apr 2025
The comment was uploaded in the form of a supplement: https://egusphere.copernicus.org/preprints/2025/egusphere-2025-501/egusphere-2025-501-AC3-supplement.pdf
- AC4: 'Reply on AC3', Junhyeong Seo, 30 Apr 2025
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EC1: 'Reply on AC1', Huixiang Xie, 19 Apr 2025
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AC1: 'Reply on RC1', Junhyeong Seo, 02 Apr 2025
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RC2: 'Comment on egusphere-2025-501', Anonymous Referee #2, 08 Apr 2025
Seo et al. present a very useful study on how the preprocessing, storage and filtration of seawater samples for FDOM measurements affects the results. This work has great value to the community, and somewhat suprising such studies have not been (to my knowledge) conducted to this degree. It's well written and concise. I recommned this is published with minor revision, which are largely to improve the clarity of presentations and provide the reader with some more useful details, as per below.
In the attached annotated ms. provided several comments and suggestions, which largely would improve the clarity of presentation. For the reader its much beneficial if some more details are given on several aspects of the methods used. Some key points include;
Provide the reader with the exact make and type of membrane filter used. Please also make sure the diameter of the filters is noted, since the volumes used for washing might be depedent on the filter size (surface area), and thus the reader needs to know how the used washing volumes relate to filter surface area.
Provide clearly (in a table), the initial concentrations of CDOM and FDOM in the filtered and unfiltered samples. This heklps the reader judge the type of samples that have been used.
It would be also valuble to show whether CDOM changed over the same period of time, as this could affect the inner filter correction.
Any ancillary data that could tell about particle loading (e.g. CHLA, POC, etc.) in the samples used in the study would be also very valuable for the reader to evaluate how particle loading might ahve affected the results obtained.
When in the field, sometimes neither distilled water nor HCL is available or practical to use, then the procedure would be to use sample water to pre-rinse the filter. Would this not be as efficient as using either of the above?
Several technical comments/suggestions are in the attached annotated ms.
- AC2: 'Reply on RC2', Junhyeong Seo, 08 Apr 2025
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AC5: 'Reply on RC2', Junhyeong Seo, 30 Apr 2025
The comment was uploaded in the form of a supplement: https://egusphere.copernicus.org/preprints/2025/egusphere-2025-501/egusphere-2025-501-AC5-supplement.pdf
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