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Influence of Organic Solvents and β-cyclodextrins on Capillary Zone Electrophoresis Separation of Five Biogenic Amines and Two B Vitamins Cover

Influence of Organic Solvents and β-cyclodextrins on Capillary Zone Electrophoresis Separation of Five Biogenic Amines and Two B Vitamins

European Pharmaceutical Journal
Volume 69 (2022): Issue 1 (January 2022)
By: M. Matuskova,  I. Cizmarova,  P. Chalova,  O. Stefanik,  A. Horniakova,  P. Mikus and  J. Piestansky  
Open Access
|Aug 2022

Figures & Tables

Figure 1

Optimisation of the background electrolyte (BGE) composition. 1 = thiamine hydrochloride (THI), 2 = tyramine hydrochloride (TYR), 3 = dopamine hydrochloride (DOP), 4 = 5-hydroxytryptamine hydrochloride (5-HT), 5 = noradrenaline hydrochloride (NOR), 6 = adrenaline hydrochloride (ADR), 7 = pyridoxine hydrochloride (PYR). Concentration of the analytes in the sample was 10 μg/mL−1. Applied separation current was 50 μA. For more operation conditions see the part Instrumentation.
Optimisation of the background electrolyte (BGE) composition. 1 = thiamine hydrochloride (THI), 2 = tyramine hydrochloride (TYR), 3 = dopamine hydrochloride (DOP), 4 = 5-hydroxytryptamine hydrochloride (5-HT), 5 = noradrenaline hydrochloride (NOR), 6 = adrenaline hydrochloride (ADR), 7 = pyridoxine hydrochloride (PYR). Concentration of the analytes in the sample was 10 μg/mL−1. Applied separation current was 50 μA. For more operation conditions see the part Instrumentation.

Figure 2

Effect of organic solvent addition into the background electrolyte (BGE) on the simultaneous separation of five biogenic amines and two B vitamins. (a) Addition of methanol (MeOH); (b) addition of acetonitrile (ACN); (c) addition of isopropanol (IP); (d) addition of tetrahydrofuran (THF). BGE = 25 mM GABA + 50 mM HAc + 0.1% mHEC. Concentration of the analytes in the sample was 10 μg/mL−1. Applied separation current was 50 μA. 1 = thiamine hydrochloride (THI), 2 = tyramine hydrochloride (TYR), 3 = dopamine hydrochloride (DOP), 4 = 5-hydroxytryptamine hydrochloride (5-HT), 5 = noradrenaline hydrochloride (NOR), 6 = adrenaline hydrochloride (ADR), 7 = pyridoxine hydrochloride (PYR). For more operation conditions, see the “Instrumentation” section.
Effect of organic solvent addition into the background electrolyte (BGE) on the simultaneous separation of five biogenic amines and two B vitamins. (a) Addition of methanol (MeOH); (b) addition of acetonitrile (ACN); (c) addition of isopropanol (IP); (d) addition of tetrahydrofuran (THF). BGE = 25 mM GABA + 50 mM HAc + 0.1% mHEC. Concentration of the analytes in the sample was 10 μg/mL−1. Applied separation current was 50 μA. 1 = thiamine hydrochloride (THI), 2 = tyramine hydrochloride (TYR), 3 = dopamine hydrochloride (DOP), 4 = 5-hydroxytryptamine hydrochloride (5-HT), 5 = noradrenaline hydrochloride (NOR), 6 = adrenaline hydrochloride (ADR), 7 = pyridoxine hydrochloride (PYR). For more operation conditions, see the “Instrumentation” section.

Figure 3

Effect of simultaneous addition of tetrahydrofuran (THF) and isopropanol (IP) into the background electrolyte (BGE) on the separation of five biogenic amines and two B vitamins. BGE = 25 mM GABA + 50 mM HAc + 0.1% m-HEC. Concentration of the analytes in the sample was 10 μg/mL−1. Applied separation current was 50 μA. 1 = thiamine hydrochloride (THI), 2 = tyramine hydrochloride (TYR), 3 = dopamine hydrochloride (DOP), 4 = 5-hydroxytryptamine hydrochloride (5-HT), 5 = noradrenaline hydrochloride (NOR), 6 = adrenaline hydrochloride (ADR), 7 = pyridoxine hydrochloride (PYR). For other operation conditions, see the “Instrumentation” section.
Effect of simultaneous addition of tetrahydrofuran (THF) and isopropanol (IP) into the background electrolyte (BGE) on the separation of five biogenic amines and two B vitamins. BGE = 25 mM GABA + 50 mM HAc + 0.1% m-HEC. Concentration of the analytes in the sample was 10 μg/mL−1. Applied separation current was 50 μA. 1 = thiamine hydrochloride (THI), 2 = tyramine hydrochloride (TYR), 3 = dopamine hydrochloride (DOP), 4 = 5-hydroxytryptamine hydrochloride (5-HT), 5 = noradrenaline hydrochloride (NOR), 6 = adrenaline hydrochloride (ADR), 7 = pyridoxine hydrochloride (PYR). For other operation conditions, see the “Instrumentation” section.

Figure 4

Effect of CE-β-CD addition into the background electrolyte (BGE) on the simultaneous separation of five biogenic amines and two B vitamins. (a) Addition of CE-β-CD at various concentration levels; (b) addition of CE-β-CD at 1 mg/mL−1 concentration level and isopropanol; (c) addition of CE-β-CD at 2 mg/mL−1 concentration level and isopropanol; (d) addition of CE-β-CD at 2.5 mg/mL−1 concentration level and isopropanol. BGE = 25 mM GABA + 50 mM HAc + 0.1% mHEC. Concentration of the analytes in the sample was 10 μg/mL−1. Applied separation current was 50 μA. 1 = thiamine hydrochloride (THI), 2 = tyramine hydrochloride (TYR), 3 = dopamine hydrochloride (DOP), 4 = 5-hydroxytryptamine hydrochloride (5-HT), 5 = noradrenaline hydrochloride (NOR), 6 = adrenaline hydrochloride (ADR), 7 = pyridoxine hydrochloride (PYR). For more operation conditions, see the “Instrumentation” section.
Effect of CE-β-CD addition into the background electrolyte (BGE) on the simultaneous separation of five biogenic amines and two B vitamins. (a) Addition of CE-β-CD at various concentration levels; (b) addition of CE-β-CD at 1 mg/mL−1 concentration level and isopropanol; (c) addition of CE-β-CD at 2 mg/mL−1 concentration level and isopropanol; (d) addition of CE-β-CD at 2.5 mg/mL−1 concentration level and isopropanol. BGE = 25 mM GABA + 50 mM HAc + 0.1% mHEC. Concentration of the analytes in the sample was 10 μg/mL−1. Applied separation current was 50 μA. 1 = thiamine hydrochloride (THI), 2 = tyramine hydrochloride (TYR), 3 = dopamine hydrochloride (DOP), 4 = 5-hydroxytryptamine hydrochloride (5-HT), 5 = noradrenaline hydrochloride (NOR), 6 = adrenaline hydrochloride (ADR), 7 = pyridoxine hydrochloride (PYR). For more operation conditions, see the “Instrumentation” section.

Figure 5

Effect of HP-β-CD addition into the background electrolyte (BGE) on the simultaneous separation of five biogenic amines and two B vitamins. (a) Addition of HP-β-CD at various concentration levels; (b) addition of HP-β-CD at 5 mg/mL−1 concentration level and isopropanol; (c) addition of HP-β-CD at 10 mg/mL−1 concentration level and isopropanol; (d) addition of HP-β-CD at 15 mg/mL−1 concentration level and isopropanol. BGE = 25 mM GABA + 50 mM HAc + 0.1% mHEC. Concentration of the analytes in the sample was 10 μg/mL−1. Applied separation current was 50 μA. 1 = thiamine hydrochloride (THI), 2 = tyramine hydrochloride (TYR), 3 = dopamine hydrochloride (DOP), 4 = 5-hydroxytryptamine hydrochloride (5-HT), 5 = noradrenaline hydrochloride (NOR), 6 = adrenaline hydrochloride (ADR), 7 = pyridoxine hydrochloride (PYR). For more operation conditions, see the “Instrumentation” section.
Effect of HP-β-CD addition into the background electrolyte (BGE) on the simultaneous separation of five biogenic amines and two B vitamins. (a) Addition of HP-β-CD at various concentration levels; (b) addition of HP-β-CD at 5 mg/mL−1 concentration level and isopropanol; (c) addition of HP-β-CD at 10 mg/mL−1 concentration level and isopropanol; (d) addition of HP-β-CD at 15 mg/mL−1 concentration level and isopropanol. BGE = 25 mM GABA + 50 mM HAc + 0.1% mHEC. Concentration of the analytes in the sample was 10 μg/mL−1. Applied separation current was 50 μA. 1 = thiamine hydrochloride (THI), 2 = tyramine hydrochloride (TYR), 3 = dopamine hydrochloride (DOP), 4 = 5-hydroxytryptamine hydrochloride (5-HT), 5 = noradrenaline hydrochloride (NOR), 6 = adrenaline hydrochloride (ADR), 7 = pyridoxine hydrochloride (PYR). For more operation conditions, see the “Instrumentation” section.

Accuracy and precision data obtained by the CZE-UV method_

Intra-day, n = 5
Found (μg.mL−1)RSD (%)% Nom.
Nominal (μg.mL−1)THITYRDOP5-HTNORADRPYRTHITYRDOP5-HTNORADRPYRTHITYRDOP5-HTNORADRPYR
0.50.54--0.43--0.568.2--10.7--15.5108.1--86.8--112.1
11.09--1.02--0.9913.1--8.2--10.6108.5--101.6--99.4
2.52.772.642.072.422.952.842.5911.08.615.14.28.94.45.4110.7105.582.997.0117.8113.5103.7
54.975.004.955.064.974.995.103.58.27.71.913.43.33.899.4100.198.9101.299.599.8101.9
109.239.2110.219.779.719.549.221.21.58.02.72.34.81.792.392.1102.197.797.195.492.2
2525.4725.9926.1525.5124.0825.1125.011.01.66.50.41.20.41.7101.9104.0104.6102.096.3100.5100.0
5049.9149.6549.4249.7950.4850.0250.100.42.13.41.22.64.00.599.899.398.999.6101.0100.1100.2

Selected operation and validation parameters of the proposed CZE-UV method_

THITYRDOP5-HTNORADRPYR
tm (min)12.0912.9514.0914.6015.0615.7318.44
RSDtm (%), n=60.240.020.110.120.100.070.21
RSDarea (%), n=61.824.308.215.474.764.271.67
Calibration equationy=104.03x + 77.112y=15.15x − 3.721y=9.64x + 8.563y=44.70x + 11.626y=7.27x − 8.347y=12.43x − 5.641y=38.48x − 3.648
r20.99940.99910.99570.99960.99240.99790.9995
Linear range0.5 – 502.5 – 502.5 – 500.5 – 502.5 – 502.5 – 500.5 – 50
LOD (μg.mL−1)0.151.251.250.251.251.250.25
LOQ (μg.mL−1)0.52.52.50.52.52.50.5
N31729287532781328162298802232727182
R 2.993.571.471.321.766.27

Stability study of the analytes performed under various storage conditions_

% difference from the initial concentration, n = 5
20°C4°C−20°C−80°C
c (μg.mL−1)2.515302.515302.515302.51530
THI−8.60.70.6−0.9−4.4−1.30.26−2.43.80.50.20.2
TYR−13.86.5−2.49.88.1−4.23.114.52.51.01.4−4.6
DOP−9.4−0.60.42.4−2.60.33.3−2.44.2−8.6−0.71.0
5−HT2.40.10.74.2−1.21.20.50.72.60.43.31.6
NOR2.02.76.3−2.8−0.49.1−5.01.52.7−14.66.55.0
ARD−1.11.42.4−1.0−7.1−1.63.32.72.0−7.07.52.3
PYR1.8−1.2−3.43.1−9.0−1.0−2.8−10.7−1.40.7−8.1−0.6
DOI: https://doi.org/10.2478/afpuc-2022-0012 | Journal eISSN: 2453-6725
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Language: English
Page range: 43 - 53
Submitted on: Jan 14, 2022
Accepted on: May 30, 2022
Published on: Aug 6, 2022
Published by: Comenius University in Bratislava, Faculty of Pharmacy
In partnership with: Paradigm Publishing Services
Publication frequency: 2 issues per year
Keywords:
biogenic amines,
thiamine,
pyridoxine,
capillary zone electrophoresis,
cyclodextrins,
organic solvents
Related subjects:
Pharmacy,
Pharmacy, other

© 2022 M. Matuskova, I. Cizmarova, P. Chalova, O. Stefanik, A. Horniakova, P. Mikus, J. Piestansky, published by Comenius University in Bratislava, Faculty of Pharmacy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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