Preanalytical and analytical challenges in gas chromatographic determination of cholesterol synthesis and absorption markers
Authorized Users Only
2018
Authors
Gojković, Tamara
Vladimirov, Sandra

Spasojević-Kalimanovska, Vesna

Zeljković, Aleksandra

Vekić, Jelena

Arsenijević, Jelena

Đuričić, Ivana

Šobajić, Slađana

Jelić-Ivanović, Zorana

Article (Published version)

Metadata
Show full item recordAbstract
Introduction: Cholesterol homeostasis disruption contributes to the development of different pathologies. Non cholesterol sterols (NCSs) serve as cholesterol synthesis markers (desmosterol and lathosterol), and cholesterol absorption surrogate markers (campesterol, stigmasterol and beta-sitosterol). The study aimed to resolve certain new pre-analytical and analytical problems and ensure a reliable and validated method. Materials and methods: Method optimization, validation and stability studies were executed in human serum and plasma. Freeze-thaw cycles were done with and without antioxidant. Gas chromatography-mass spectrometer (GC-MS) was used for NCSs confirmation and plasticizer identification, while GC-flame ionization detector (GC-FID) was used for NCSs quantitation. Results: Infra-and inter-assay variabilities for all NCSs were 2.75-9.55% and 5.80-7.75% for plasma and 3.10-5.72% and 3.05-10.92% for serum, respectively. Recovery studies showed satisfactory percentage errors for a...ll NCSs: 93.4-105.7% in plasma and 87.5-106.9 in serum. Derivatized samples were stable up to 7 days at 20 degrees C and derivatization yield was affected by presence of plasticizers. Fatty acid amids were identified as interfering plastic leachates. Statistically different NCSs concentrations were observed after the 1st freeze-thaw cycle, in antioxidant-free samples, and after the 4th cycle in antioxidant-enriched samples. Conclusions: All of the in-house procedures proved to be useful for minimizing the preanalytical and analytical variations, as proven by the validation results.
Keywords:
Method validation / Sterols / Stability / Freeze-thaw / PlasticizersSource:
Clinica Chimica Acta, 2018, 478, 74-81Publisher:
- Elsevier Science BV, Amsterdam
Funding / projects:
DOI: 10.1016/j.cca.2017.12.032
ISSN: 0009-8981
PubMed: 29274328
WoS: 000425070000013
Scopus: 2-s2.0-85039173890
Collections
Institution/Community
PharmacyTY - JOUR AU - Gojković, Tamara AU - Vladimirov, Sandra AU - Spasojević-Kalimanovska, Vesna AU - Zeljković, Aleksandra AU - Vekić, Jelena AU - Arsenijević, Jelena AU - Đuričić, Ivana AU - Šobajić, Slađana AU - Jelić-Ivanović, Zorana PY - 2018 UR - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3185 AB - Introduction: Cholesterol homeostasis disruption contributes to the development of different pathologies. Non cholesterol sterols (NCSs) serve as cholesterol synthesis markers (desmosterol and lathosterol), and cholesterol absorption surrogate markers (campesterol, stigmasterol and beta-sitosterol). The study aimed to resolve certain new pre-analytical and analytical problems and ensure a reliable and validated method. Materials and methods: Method optimization, validation and stability studies were executed in human serum and plasma. Freeze-thaw cycles were done with and without antioxidant. Gas chromatography-mass spectrometer (GC-MS) was used for NCSs confirmation and plasticizer identification, while GC-flame ionization detector (GC-FID) was used for NCSs quantitation. Results: Infra-and inter-assay variabilities for all NCSs were 2.75-9.55% and 5.80-7.75% for plasma and 3.10-5.72% and 3.05-10.92% for serum, respectively. Recovery studies showed satisfactory percentage errors for all NCSs: 93.4-105.7% in plasma and 87.5-106.9 in serum. Derivatized samples were stable up to 7 days at 20 degrees C and derivatization yield was affected by presence of plasticizers. Fatty acid amids were identified as interfering plastic leachates. Statistically different NCSs concentrations were observed after the 1st freeze-thaw cycle, in antioxidant-free samples, and after the 4th cycle in antioxidant-enriched samples. Conclusions: All of the in-house procedures proved to be useful for minimizing the preanalytical and analytical variations, as proven by the validation results. PB - Elsevier Science BV, Amsterdam T2 - Clinica Chimica Acta T1 - Preanalytical and analytical challenges in gas chromatographic determination of cholesterol synthesis and absorption markers VL - 478 SP - 74 EP - 81 DO - 10.1016/j.cca.2017.12.032 ER -
@article{ author = "Gojković, Tamara and Vladimirov, Sandra and Spasojević-Kalimanovska, Vesna and Zeljković, Aleksandra and Vekić, Jelena and Arsenijević, Jelena and Đuričić, Ivana and Šobajić, Slađana and Jelić-Ivanović, Zorana", year = "2018", abstract = "Introduction: Cholesterol homeostasis disruption contributes to the development of different pathologies. Non cholesterol sterols (NCSs) serve as cholesterol synthesis markers (desmosterol and lathosterol), and cholesterol absorption surrogate markers (campesterol, stigmasterol and beta-sitosterol). The study aimed to resolve certain new pre-analytical and analytical problems and ensure a reliable and validated method. Materials and methods: Method optimization, validation and stability studies were executed in human serum and plasma. Freeze-thaw cycles were done with and without antioxidant. Gas chromatography-mass spectrometer (GC-MS) was used for NCSs confirmation and plasticizer identification, while GC-flame ionization detector (GC-FID) was used for NCSs quantitation. Results: Infra-and inter-assay variabilities for all NCSs were 2.75-9.55% and 5.80-7.75% for plasma and 3.10-5.72% and 3.05-10.92% for serum, respectively. Recovery studies showed satisfactory percentage errors for all NCSs: 93.4-105.7% in plasma and 87.5-106.9 in serum. Derivatized samples were stable up to 7 days at 20 degrees C and derivatization yield was affected by presence of plasticizers. Fatty acid amids were identified as interfering plastic leachates. Statistically different NCSs concentrations were observed after the 1st freeze-thaw cycle, in antioxidant-free samples, and after the 4th cycle in antioxidant-enriched samples. Conclusions: All of the in-house procedures proved to be useful for minimizing the preanalytical and analytical variations, as proven by the validation results.", publisher = "Elsevier Science BV, Amsterdam", journal = "Clinica Chimica Acta", title = "Preanalytical and analytical challenges in gas chromatographic determination of cholesterol synthesis and absorption markers", volume = "478", pages = "74-81", doi = "10.1016/j.cca.2017.12.032" }
Gojković, T., Vladimirov, S., Spasojević-Kalimanovska, V., Zeljković, A., Vekić, J., Arsenijević, J., Đuričić, I., Šobajić, S.,& Jelić-Ivanović, Z.. (2018). Preanalytical and analytical challenges in gas chromatographic determination of cholesterol synthesis and absorption markers. in Clinica Chimica Acta Elsevier Science BV, Amsterdam., 478, 74-81. https://doi.org/10.1016/j.cca.2017.12.032
Gojković T, Vladimirov S, Spasojević-Kalimanovska V, Zeljković A, Vekić J, Arsenijević J, Đuričić I, Šobajić S, Jelić-Ivanović Z. Preanalytical and analytical challenges in gas chromatographic determination of cholesterol synthesis and absorption markers. in Clinica Chimica Acta. 2018;478:74-81. doi:10.1016/j.cca.2017.12.032 .
Gojković, Tamara, Vladimirov, Sandra, Spasojević-Kalimanovska, Vesna, Zeljković, Aleksandra, Vekić, Jelena, Arsenijević, Jelena, Đuričić, Ivana, Šobajić, Slađana, Jelić-Ivanović, Zorana, "Preanalytical and analytical challenges in gas chromatographic determination of cholesterol synthesis and absorption markers" in Clinica Chimica Acta, 478 (2018):74-81, https://doi.org/10.1016/j.cca.2017.12.032 . .