Quantification of total content of non-esterified fatty acids bound to human serum albumin
Samo za registrovane korisnike
2016
Autori
Pavićević, Ivan D.Jovanović, Vesna B.
Takić, Marija
Aćimović, Jelena M.
Penezić, Ana Z.
Mandić, Ljuba M.
Članak u časopisu (Objavljena verzija)
Metapodaci
Prikaz svih podataka o dokumentuApstrakt
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.
Ključne reči:
Fatty acid bound to HSA / Fatty acids/HSA ratio / TLC quantification / Flatbed scanner and digital image analysisIzvor:
Journal of Pharmaceutical & Biomedical Analysis, 2016, 129, 43-49Izdavač:
- Elsevier Science Bv, Amsterdam
Finansiranje / projekti:
- Alergeni, antitela, enzimi i mali fiziološki značajni molekuli: dizajn, struktura, funkcija i značaj (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
Institucija/grupa
Institut za medicinska istraživanjaTY - JOUR AU - Pavićević, Ivan D. AU - Jovanović, Vesna B. AU - Takić, Marija AU - Aćimović, Jelena M. AU - Penezić, Ana Z. AU - Mandić, Ljuba M. PY - 2016 UR - http://rimi.imi.bg.ac.rs/handle/123456789/698 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. PB - Elsevier Science Bv, Amsterdam T2 - Journal of Pharmaceutical & Biomedical Analysis T1 - Quantification of total content of non-esterified fatty acids bound to human serum albumin EP - 49 SP - 43 VL - 129 DO - 10.1016/j.jpba.2016.06.043 ER -
@article{ author = "Pavićević, Ivan D. and Jovanović, Vesna B. and Takić, Marija 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.", publisher = "Elsevier Science Bv, Amsterdam", journal = "Journal of Pharmaceutical & Biomedical Analysis", title = "Quantification of total content of non-esterified fatty acids bound to human serum albumin", pages = "49-43", volume = "129", doi = "10.1016/j.jpba.2016.06.043" }
Pavićević, I. D., Jovanović, V. B., Takić, 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 & Biomedical Analysis Elsevier Science Bv, Amsterdam., 129, 43-49. https://doi.org/10.1016/j.jpba.2016.06.043
Pavićević ID, Jovanović VB, Takić M, Aćimović JM, Penezić AZ, Mandić LM. Quantification of total content of non-esterified fatty acids bound to human serum albumin. in Journal of Pharmaceutical & Biomedical Analysis. 2016;129:43-49. doi:10.1016/j.jpba.2016.06.043 .
Pavićević, Ivan D., Jovanović, Vesna B., Takić, Marija, 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 & Biomedical Analysis, 129 (2016):43-49, https://doi.org/10.1016/j.jpba.2016.06.043 . .