HPLC-MS/MS测定大鼠血浆中杨芽黄素的含量

doi:10.3976/j.issn.1002-4026.2022.04.004

Abstract

Abstract:

The aim was to develop a method using high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) for the quantitative analysis of tectochrysin in rat plasma and evaluate the pharmacokinetics of tectochrysin in rats after intragastric administration. First, plasma samples were precipitated using acetonitrile reagent, and a 5 μL supernatant was injected into system for analysis. Phellopterin was used as the internal standard to monitor the concentrations of tectochrysin in rat plasma via HPLC-MS/MS. Zorbax Eclipse Plus C₁₈ column (2.1 mm × 50 mm, 1.8 μm) was used, and the mobile phase was composed of an aqueous solution (60:40, V/V) of 0.1% acetonitrile and formic acid (isocratic elution). Electrospray ionization was used in the positive ion mode, and selected reaction monitoring was performed at mass-to-charge ratio (m/z) 268.8→225.9 (tectochrysin) and m/z 300.9→233.0 (internal standard). The calibration curve was linear in the range of 0.5~200 ng/mL. The relative standard deviation of inter and intraday precisions was 3.28%~13.12%, and relative error of accuracy was -6.38%~7.60%. Matrix effect was 98.43%~103.57%, and extraction recovery rate was 93.36%~96.31%. The assay was found to be useful for determining the concentration and evaluating the pharmacokinetics of tectochrysin in rat plasma.

Key words: tectochrysin, HPLC-MS/MS, rat plasma, matrix effect, pharmacokinetics

CLC Number:

R285

DENG Zhi-Peng, LI Jing, ZHAO Pan, SONG Jia. Determining the concentration of tectochrysin in rat plasma using HPLC-MS/MS[J].Shandong Science, 2022, 35(4): 21-27.

Figures/Tables 8

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Fig.2

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Table 1

Table 2

Determination of the recovery rate and ME of tectochrysin in rat plasma using LC-MS/MS (n=6)"

样品质量浓度/ (ng·mL^-1)	提取回收率/%		基质效应/%
样品质量浓度/ (ng·mL^-1)	$x ¯$ ±s	δ_RSD	$x ¯$ ±s	δ_RSD
1.5	95.9±5.4	5.6	103.6±3.3	3.2
15.0	96.3±2.7	2.8	98.4±5.5	5.6
180.0	93.4±6.6	7.1	100.0±6.1	6.1

Table 2

Table 3

Fig.4

Table 4

References 12

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样品质量浓度/ (ng·mL^-1)	$\bar{x}$±s/ (ng·mL^-1)		δ_RSD/%		E_R/%
样品质量浓度/ (ng·mL^-1)	日内	日间	日内	日间	日内	日间
1.5	1.4±0.1	1.5±0.1	4.8	9.2	-6.8	1.6
15.0	16.1±2.1	14.4±1.1	13.1	7.4	7.6	-3.9
180.0	184.7±17.8	171.3±5.6	9.6	3.3	2.6	-4.8

处置条件	1.5 ng/mL		15 ng/mL		180 ng/mL
处置条件	δ_RSD	E_R	δ_RSD	E_R	δ_RSD	E_R
室温放置8 h	6.3	3.2	5.3	-7.5	1.1	-9.7
-20 ℃长期冷冻30 d	1.6	-3.4	6.6	-2.4	1.0	-5.1
反复冻融3次	5.8	8.4	12.0	-13.8	2.1	3.5
进样室放置6 h	0.9	2.6	10.2	-9.5	3.0	6.1

参数	单位	杨芽黄素给药量
参数	单位	5 mg/kg	10 mg/kg	20 mg/kg
AUC_0~_t	μg/(L·h)	42.6±14.6	96.1±45.4	187.1±87.6
AUC_0~∞	μg/(L·h)	44.9±14.8	98.8±44.8	191.3±86.6
MRT	h	2.5±0.7	3.8±1.6	3.8±1.3
t_1/2	h	2.4±0.8	3.7±2.1	3.4±1.1
C_max	μg/(L·h)	25.4±5.5	41.4±9.7	74.5±15.4
T_max	h	0.33±0.12	0.37±0.14	0.30±0.14

Determining the concentration of tectochrysin in rat plasma using HPLC-MS/MS

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