Quantification of total content of non-esterified fatty acids bound to human serum albumin
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2016
Authors
Pavićević, Ivan D.Jovanović, Vesna B.
Takić, Marija M.
Aćimović, Jelena M.
Penezić, Ana Z.
Mandić, Ljuba M.
Article (Published version)
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Non-esterified fatty acids bound to the human serum albumin (HSA) contribute to several HSAs properties of special concern in pathologies, for instance to the reactivity of the free HSA-Cys34 thiol group (important antioxidative thiol pool in plasma), and to the affinity for binding of molecules and ions (for example cobalt as a prominent biomarker in heart ischemia). Therefore, the method for determination of FAs bound to HSA was developed. FAs were released from HSA (previously isolated from serum by ammonium sulfate precipitation) using acidic copper(II) sulfate in phosphoric acid, extracted by n-heptane-chloroform (4:1, v/v) mixture, spotted on TL silica-gel and then developed with n-heptane-chloroform-acetic acid (5:3:03, v/v/v). Common office flatbed scanner and software solution for densitometric image analysis, developed in R, were used. The linearity of calibration curve in concentration range from 0.1 to 5.0 mmol/L stearic acid was achieved. The method was proved to be precis...e (with RSD of 1.4-4.7%) and accurate. Accuracy was examined by standard addition method (recoveries 97.2-102.5%) and by comparison to results of GC. The method is sample saving, technically less demanding, and cheap, and therefore suitable for determination of FAs/HSA ratio when elevated concentrations of free FAs are reliable diagnostic/risk parameter of pathological states. (C) 2016 Elsevier B.V. All rights reserved.
Keywords:
Fatty acid bound to HSA / Fatty acids/HSA ratio / TLC quantification / Flatbed scanner and digital image analysisSource:
Journal of Pharmaceutical and Biomedical Analysis, 2016, 129, 43-49Publisher:
- Elsevier Science Bv, Amsterdam
Funding / projects:
- Allergens, antibodies, enzymes and small physiologically important molecules: design, structure, function and relevance (RS-MESTD-Basic Research (BR or ON)-172049)
DOI: 10.1016/j.jpba.2016.06.043
ISSN: 0731-7085
PubMed: 27394177
WoS: 000393847600007
Scopus: 2-s2.0-84977659953
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Institution/Community
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Pavićević, Ivan D. AU - Jovanović, Vesna B. AU - Takić, Marija M. AU - Aćimović, Jelena M. AU - Penezić, Ana Z. AU - Mandić, Ljuba M. PY - 2016 UR - https://cherry.chem.bg.ac.rs/handle/123456789/2399 AB - Non-esterified fatty acids bound to the human serum albumin (HSA) contribute to several HSAs properties of special concern in pathologies, for instance to the reactivity of the free HSA-Cys34 thiol group (important antioxidative thiol pool in plasma), and to the affinity for binding of molecules and ions (for example cobalt as a prominent biomarker in heart ischemia). Therefore, the method for determination of FAs bound to HSA was developed. FAs were released from HSA (previously isolated from serum by ammonium sulfate precipitation) using acidic copper(II) sulfate in phosphoric acid, extracted by n-heptane-chloroform (4:1, v/v) mixture, spotted on TL silica-gel and then developed with n-heptane-chloroform-acetic acid (5:3:03, v/v/v). Common office flatbed scanner and software solution for densitometric image analysis, developed in R, were used. The linearity of calibration curve in concentration range from 0.1 to 5.0 mmol/L stearic acid was achieved. The method was proved to be precise (with RSD of 1.4-4.7%) and accurate. Accuracy was examined by standard addition method (recoveries 97.2-102.5%) and by comparison to results of GC. The method is sample saving, technically less demanding, and cheap, and therefore suitable for determination of FAs/HSA ratio when elevated concentrations of free FAs are reliable diagnostic/risk parameter of pathological states. (C) 2016 Elsevier B.V. All rights reserved. PB - Elsevier Science Bv, Amsterdam T2 - Journal of Pharmaceutical and Biomedical Analysis T1 - Quantification of total content of non-esterified fatty acids bound to human serum albumin VL - 129 SP - 43 EP - 49 DO - 10.1016/j.jpba.2016.06.043 ER -
@article{ author = "Pavićević, Ivan D. and Jovanović, Vesna B. and Takić, Marija M. and Aćimović, Jelena M. and Penezić, Ana Z. and Mandić, Ljuba M.", year = "2016", abstract = "Non-esterified fatty acids bound to the human serum albumin (HSA) contribute to several HSAs properties of special concern in pathologies, for instance to the reactivity of the free HSA-Cys34 thiol group (important antioxidative thiol pool in plasma), and to the affinity for binding of molecules and ions (for example cobalt as a prominent biomarker in heart ischemia). Therefore, the method for determination of FAs bound to HSA was developed. FAs were released from HSA (previously isolated from serum by ammonium sulfate precipitation) using acidic copper(II) sulfate in phosphoric acid, extracted by n-heptane-chloroform (4:1, v/v) mixture, spotted on TL silica-gel and then developed with n-heptane-chloroform-acetic acid (5:3:03, v/v/v). Common office flatbed scanner and software solution for densitometric image analysis, developed in R, were used. The linearity of calibration curve in concentration range from 0.1 to 5.0 mmol/L stearic acid was achieved. The method was proved to be precise (with RSD of 1.4-4.7%) and accurate. Accuracy was examined by standard addition method (recoveries 97.2-102.5%) and by comparison to results of GC. The method is sample saving, technically less demanding, and cheap, and therefore suitable for determination of FAs/HSA ratio when elevated concentrations of free FAs are reliable diagnostic/risk parameter of pathological states. (C) 2016 Elsevier B.V. All rights reserved.", publisher = "Elsevier Science Bv, Amsterdam", journal = "Journal of Pharmaceutical and Biomedical Analysis", title = "Quantification of total content of non-esterified fatty acids bound to human serum albumin", volume = "129", pages = "43-49", doi = "10.1016/j.jpba.2016.06.043" }
Pavićević, I. D., Jovanović, V. B., Takić, M. M., Aćimović, J. M., Penezić, A. Z.,& Mandić, L. M.. (2016). Quantification of total content of non-esterified fatty acids bound to human serum albumin. in Journal of Pharmaceutical and Biomedical Analysis Elsevier Science Bv, Amsterdam., 129, 43-49. https://doi.org/10.1016/j.jpba.2016.06.043
Pavićević ID, Jovanović VB, Takić MM, Aćimović JM, Penezić AZ, Mandić LM. Quantification of total content of non-esterified fatty acids bound to human serum albumin. in Journal of Pharmaceutical and Biomedical Analysis. 2016;129:43-49. doi:10.1016/j.jpba.2016.06.043 .
Pavićević, Ivan D., Jovanović, Vesna B., Takić, Marija M., Aćimović, Jelena M., Penezić, Ana Z., Mandić, Ljuba M., "Quantification of total content of non-esterified fatty acids bound to human serum albumin" in Journal of Pharmaceutical and Biomedical Analysis, 129 (2016):43-49, https://doi.org/10.1016/j.jpba.2016.06.043 . .