高效液相色谱法测定小鼠血浆中苯并三唑类紫外线吸收剂UV-327和UV-328

doi:10.19723/j.issn.1671-167X.2020.03.030

Abstract

Abstract:

Objective: To establish a high performance liquid chromatography (HPLC) method for the determination of ultraviolet (UV) absorbers UV-327 and UV-328 in mouse plasma.Methods: N-hexane-acetone (volume ratio 1 ∶1) was added to a mouse plasma sample as the extraction solvent for vortex extraction, and the supernatant was dried at 50 ℃ with nitrogen. Thereafter the residue was redissolved with methanol, centrifuged and filtered. The separation was performed on a Waters Symmetry^?C₁₈ column (250 mm×4.6 mm, 5 μm), and the concentrations of UV-327 and UV-328 in the mouse plasma were determined by HPLC with an UV detector. The elution was isocratic at a flow rate of 1.0 mL/min with a mobile phase composed of 100% methanol, and the UV detection wavelength was 340 nm. The retention time was used for qualitative analysis, and the internal standard method was used for quantitative analysis using UV-320 as the internal standard. Results: The calibration curves of UV-327 and UV-328 were linear with correlation coefficients of 0.999 7 over the concentration range of 0.05 to 10.0 mg/L. The limit of detection was 0.01 mg/L, and the limit of quantitation was 0.03 mg/L. The average recoveries at low, medium and high three concentrations (0.50, 1.00, 2.00 mg/L) in the mouse plasma were 91.7%-101.0% for UV-327, and 97.5%-103.9% for UV-328. The intra-day precisions (n=6) of UV-327 were 2.9%-6.6%, and 2.7%-7.4% for UV-328. The inter-day precisions (n=3) of UV-327 were 6.0%-9.3%, and 6.6%-8.6% for UV-328. The extraction recoveries of UV-327 were 98.8%-103.8%, and 99.8%-100.9% for UV-328. The measured relative deviations of UV-327 in the mouse plasma samples placed at room temperature for 6 hours and -40 ℃ for 15 days were 0.9%-3.5% and 7.4%-15.0%, and the measured relative deviations of UV-328 were 2.0%-4.3% and 2.1%-13.8%, respectively. The mouse plasma samples could be stored at room temperature for 6 hours at least and -40 ℃ for 15 days at three spiked concentration levels.Conclusion: The method was simple and fast with high accuracy, precision and sensitivity, and could be applied to the determination of UV-327 and UV-328 in mouse plasma.

Key words: High performance liquid chromatography, UV-327, UV-328, Mouse, Plasma

CLC Number:

R194.2

Mei-qing ZHU,Rong CUI. Determination of UV-327 and UV-328 in mouse plasma by high performance liquid chromatography[J].Journal of Peking University (Health Sciences), 2020, 52(3): 591-596.

Figures/Tables 5

Figure 1

Table 1

Accuracy and precision of UV-327 and UV-328 in mice plasma"

Analyte	Spiked concentration/(mg/L)	Measured concentration/(mg/L), $x ?$ ±s	Average recovery	Intra-day precision (RSD, n=6)	Inter-day precision (RSD, n=3)
UV-327	0.50	0.46±0.04	91.7%	6.6%	9.3%
	1.00	1.01±0.08	101.0%	4.3%	7.7%
	2.00	2.00±0.12	99.9%	2.9%	6.0%
UV-328	0.50	0.49±0.04	97.5%	7.4%	8.6%
	1.00	1.04±0.07	103.9%	3.6%	6.6%
	2.00	2.01±0.14	100.8%	2.7%	6.8%

Table 1

Table 2

Table 3

Stability test results of UV-327 and UV-328 in mice plasma (n=6)"

Analyte	Spiked concentration/ (mg/L)	Instant	Room temperature for 6 h		-40 ℃ for 15 d
Analyte	Spiked concentration/ (mg/L)	Measured concentration/ (mg/L), $x ?$ ±s	Measured concentration/ (mg/L), $x ?$ ±s	Relative deviation	Measured concentration/ (mg/L), $x ?$ ±s	Relative deviation
UV-327	0.50	0.45±0.01	0.45±0.01	0.9%	0.51±0.04	13.3%
	1.00	0.95±0.04	0.94±0.01	1.1%	0.88±0.02	7.4%
	2.00	2.00±0.04	1.93±0.17	3.5%	1.70±0.04	15.0%
UV-328	0.50	0.47±0.01	0.49±0.02	4.3%	0.46±0.01	2.1%
	1.00	1.02±0.02	1.04±0.04	2.0%	0.92±0.02	9.8%
	2.00	2.10±0.05	2.03±0.13	3.3%	1.81±0.05	13.8%

Table 3

Figure 2

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Determination of UV-327 and UV-328 in mouse plasma by high performance liquid chromatography

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