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Simultaneous determination of catecholamines and polyamines in PC-12 cell extracts by micellar electrokinetic capillary chromatography with ultraviolet absorbance detection.
J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Jun 15; 805(2):281-8.JC

Abstract

A method for simultaneous determination of polyamines and catecholamines in cell extracts by micellar electrokinetic capillary chromatography with UV detection at 254 nm was established at the first time. The polyamines (putrescine, spermidine and spermine) and catecholamines (dopamine, serotonin, norepinephrine and epinephrine) were extracted from PC-12 cells and were derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. Different derivatization conditions such as temperature, ratio of derivatization reagents and incubation time were investigated to find the best reaction condition which gave the highest detection sensitivity for polyamines and catecholamines. The influence of running buffer and additives on the separation such as pH, sodium dodecyl sulfate (SDS) concentrations and various additives was also investigated. Separation was achieved within 20 min with good repeatability in a 100mM boric acid buffer containing 10mM SDS and 10mM 18-crown-6 at a pH of 9.5. The detection limit ranged from 1.0 x 10(-7) to 9.0 x 10(-7) M, which is sufficient for determination of polyamines and catecholamines in many cell extracts. This technique can be easily applied to polyamine-related anticancer drug studies or clinical follow-ups after each dosage of these anticancer drugs, since these drugs not only have great inhibition on polyamine levels in blood, but also have a large influence on catecholamine levels in blood.

Authors+Show Affiliations

Department of Chemistry, University of Missouri-Rolla, MO 65409, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15135102

Citation

Liu, Guanshu, et al. "Simultaneous Determination of Catecholamines and Polyamines in PC-12 Cell Extracts By Micellar Electrokinetic Capillary Chromatography With Ultraviolet Absorbance Detection." Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, vol. 805, no. 2, 2004, pp. 281-8.
Liu G, Chen J, Ma Y. Simultaneous determination of catecholamines and polyamines in PC-12 cell extracts by micellar electrokinetic capillary chromatography with ultraviolet absorbance detection. J Chromatogr B Analyt Technol Biomed Life Sci. 2004;805(2):281-8.
Liu, G., Chen, J., & Ma, Y. (2004). Simultaneous determination of catecholamines and polyamines in PC-12 cell extracts by micellar electrokinetic capillary chromatography with ultraviolet absorbance detection. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 805(2), 281-8.
Liu G, Chen J, Ma Y. Simultaneous Determination of Catecholamines and Polyamines in PC-12 Cell Extracts By Micellar Electrokinetic Capillary Chromatography With Ultraviolet Absorbance Detection. J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Jun 15;805(2):281-8. PubMed PMID: 15135102.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Simultaneous determination of catecholamines and polyamines in PC-12 cell extracts by micellar electrokinetic capillary chromatography with ultraviolet absorbance detection. AU - Liu,Guanshu, AU - Chen,Junnan, AU - Ma,Yinfa, PY - 2003/12/15/received PY - 2004/03/03/revised PY - 2004/03/05/accepted PY - 2004/5/12/pubmed PY - 2005/1/22/medline PY - 2004/5/12/entrez SP - 281 EP - 8 JF - Journal of chromatography. B, Analytical technologies in the biomedical and life sciences JO - J Chromatogr B Analyt Technol Biomed Life Sci VL - 805 IS - 2 N2 - A method for simultaneous determination of polyamines and catecholamines in cell extracts by micellar electrokinetic capillary chromatography with UV detection at 254 nm was established at the first time. The polyamines (putrescine, spermidine and spermine) and catecholamines (dopamine, serotonin, norepinephrine and epinephrine) were extracted from PC-12 cells and were derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. Different derivatization conditions such as temperature, ratio of derivatization reagents and incubation time were investigated to find the best reaction condition which gave the highest detection sensitivity for polyamines and catecholamines. The influence of running buffer and additives on the separation such as pH, sodium dodecyl sulfate (SDS) concentrations and various additives was also investigated. Separation was achieved within 20 min with good repeatability in a 100mM boric acid buffer containing 10mM SDS and 10mM 18-crown-6 at a pH of 9.5. The detection limit ranged from 1.0 x 10(-7) to 9.0 x 10(-7) M, which is sufficient for determination of polyamines and catecholamines in many cell extracts. This technique can be easily applied to polyamine-related anticancer drug studies or clinical follow-ups after each dosage of these anticancer drugs, since these drugs not only have great inhibition on polyamine levels in blood, but also have a large influence on catecholamine levels in blood. SN - 1570-0232 UR - https://www.unboundmedicine.com/medline/citation/15135102/Simultaneous_determination_of_catecholamines_and_polyamines_in_PC_12_cell_extracts_by_micellar_electrokinetic_capillary_chromatography_with_ultraviolet_absorbance_detection_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S157002320400220X DB - PRIME DP - Unbound Medicine ER -