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Determination of naringin in samples by external standard method
Determination of naringin in samples by external standard method
1. Experimental method
Cometro high performance liquid chromatography system;
Column: Comasil C18 250 * 4.6mm, 5μm;
Mobile phase: methanol-water = 40: 60;
UV detector wavelength: 283nm;
Flow rate: 1.0ml / min;
Sample: naringin reference substance (19.68μg / ml), sample;
Injection volume: the control sample was injected into each of 2.5μl, 4μl, 5μl, 6μl, 7.5μl 5 times, the sample was injected 5μl, the sample was mixed with the standard 5: 1 and then injected 5μl, the linear correlation and retention time Reproducibility, peak area reproducibility, recovery rate.
2. Experimental results and analysis
1. Reference substance 5μl
Channel A Results
Name
Retention Time
Area
Theoretical plates (USP)
Resolution (USP)
Asymmetry
Naringin
18.477
89103
8137
0.00000
1.09209
Attached sample volume, retention time, peak area relationship table:
Injection volume / μl
Retention time / min
Peak area / mV * min
2.5
18.386
42387
4
18.377
71244
5
18.477
89103
6
18.539
106504
7.5
18.501
134769
2. Sample 5μl
Channel A Results
Name
Retention Time
Area
Theoretical plates (USP)
Resolution (USP)
Asymmetry
Naringin
19.497
128002
8087
5.22515
1.05546
3. Sample is mixed with standard 5: 1 and injected 5μl
Channel A Results
Name
Retention Time
Area
Theoretical plates (USP)
Resolution (USP)
Asymmetry
Naringin
18.535
120336
7828
2.87114
1.03257
Three, result operation
1. Reproducibility of retention time
keep time
18.386
18.377
18.477
18.539
18.501
RSD
0.3747%
2. Peak area reproducibility investigation
Peak area
89103
89105
88953
RSD
0.0979%
3. Investigation of linear correlation and calculation of sample content
Taking the injection volume as the abscissa and the peak area as the ordinate, the linear equation is obtained:
From this, the sample contains naringin 28.08μg / ml.
4. Investigation of recovery rate:
According to the above linear equation, the mixed sample contains naringin 26.43μg / ml, and the actual true value is 26.68μg / ml, so the recovery rate is 99.06%.
4. Overview
These four investigations comprehensively examined the absorption stability of the detector, the accuracy of the pump flow rate, the accuracy of the injection valve and the injection needle, and the quality of the Comasil column.