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The determination of ϵ-amino groups in soluble and poorly soluble proteinaceous materials by a spectrophotometrie method using trinitrobenzenesulfonic acid
Abstract
A procedure using 2,4,6-trinitrobenzenesulfonic acid (TNBS) for the determination of epsilon-amino groups in soluble and poorly soluble proteinaceous materials is presented. The major modification from previous procedures is an extended TNBS reaction time to allow a stoichiometric reaction with amino groups. In addition, autoclave hydrolysis is used to assure sample dissolution for spectrophotometric measurements. The assay accuracy was evaluated by determining epsilon-amino groups of insulin and bovine albumin. The determinations differed from literature values by < or = 3.3%. The epsilon-amino group content of Type B gelatin was found to be 33.0 mol/gelatin molecule of 1000 residues and is in agreement with similar source gelatins and collagen. The coefficient of variation for determinations on all three materials was < or = 5.3%. The assay should be applicable to a broad range of proteinaceous materials.
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“For SF (Figure 1C), the main characteristic bands were observed at 1641 cm⁻¹, corresponding to C=O stretching (Amide I); 1514 cm⁻¹, associated with Amide II vibrations, including in-plane N-H bending and C-N stretching; and 1234 cm⁻¹, corresponding to Amide III, resulting from N-C stretching and C=O bending, related to β-sheet formation [27,36].”
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Metadata
- Authors
- Bubnis, W. A., & Ofner, C. M.
- Affiliations
- University of the Sciences (US)
- ORCID
- Ofner, Clyde M.
- Venue
- Analytical Biochemistry
- Publisher
- Elsevier BV
- Published
- 1992-11
- Volume
- 207
- Issue
- 1
- Pages
- 129-133
- ISSN
- 0003-2697 (ISSN-L)
- OpenAlex
- W1970226578
- Cited by
- 265
- References
- 19
- Citations by year
- 2026: 3 · 2025: 10 · 2024: 7 · 2023: 8 · 2022: 4 · 2021: 12 · 2020: 6 · 2019: 11 · 2018: 13 · 2017: 12 · 2016: 15 · 2015: 19 · 2014: 13 · 2013: 19 · 2012: 20
- FWCI
- 0.74
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Analytical Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Physical Sciences › Materials Science › Biomaterials › Collagen: Extraction and Characterization
- Life Sciences › Agricultural and Biological Sciences › Food Science › Food Nutrient Composition and Product Development
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Keywords
- epsilon-amino groups, 2,4,6-trinitrobenzene sulfonic acid, gelatin, collagen, insulin, bovine serum albumin, spectrophotometric assay
- MeSH terms
- Albumins, Amines, Animals, Cattle, Gelatin, Humans, Insulin, Molecular Weight, Proteins, Sensitivity and Specificity, Solubility, Spectrophotometry, Time Factors, Trinitrobenzenesulfonic Acid
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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