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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: ,  ,  ,  ,  ,   and    
Open Access
|Aug 2022
Figures & tables

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.

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.

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.

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.

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.

Table 1

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
R2.993.571.471.321.766.27

[i] tm – migration time, RSDtm – relative standard deviation of migration time, RSDarea – relative standard deviation of peak area, LOD – limit of detection, LOQ – limit of quantification, N – separation efficiency, R – resolution. Separation efficiency (N) was calculated according to the equation N = 5.545*(tm/w1/2)2, where tm is the migration time and w1/2 is the full width at half maximum of the peak. Resolution (R) was calculated according to the equation R = 1.18*(tmB – tmA)/(w1/2A + w1/2B), where tmA and tmB represent migration times of analyte A and B, and w1/2A and w1/2B represent the full width of the peaks A and B at their half maximum. The calibration curve is expressed by the equation y = b.x + a. RSDtm and RSDarea were calculated from the samples at LOQ concentration level. LOD and LOQ were calculated as the signal (S) to noise (N) ratios to be 3xS/N and 10xS/N, respectively.

Table 2

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
Inter-day, n = 15
Found (μg.mL−1)RSD (%)% Nom.
Nominal (μg.mL−1)THITYRDOP5-HTNORADRPYRTHITYRDOP5-HTNORADRPYRTHITYRDOP5-HTNORADRPYR
0.50.57--0.44--0.5910.7--15.0--13.4113.5--87.6--117.8
10.96--1.02--1.0014.7--11.7--8.195.7--102.3--99.6
2.52.612.602.152.423.022.982.4910.114.818.33.86.212.56.9104.5103.985.896.8120.8119.299.6
55.215.054.764.865.335.345.235.65.68.84.811.111.112.0104.3100.993.897.1106.6106.9104.6
109.729.5610.6210.119.709.189.655.74.18.84.32.67.15.697.295.6106.2101.197.091.896.5
2525.5425.4426.1925.4223.4024.7824.810.93.25.50.96.62.01.7102.2101.8104.7101.793.699.199.2
5049.7749.8649.3649.7950.7950.2250.140.42.02.71.02.43.00.999.599.798.799.6101.6100.4100.3
Table 3

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 (formerly 1338-6786) | Journal ISSN: 0301-2298
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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
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.

Volume 69 (2022): Issue 1 (January 2022)
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