Determination of 15 lipid regulators in fish meat by QuEChERS-ultra performance liquid chromatography-quadrupole/ electrostatic field orbitrap mass spectrometry

doi:10.3724/SP.J.1123.2019.10011

Abstract

Abstract:

An analytical method was established for the determination of 15 lipid regulators in fish meat by QuEChERS combined with ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap mass spectrometry (UPLC-Q/Orbitrap-MS). The samples were purified by optimized QuEChERS methods. The amounts of the adsorbent materials (primary secondary amine (PSA) (20, 60, 100, 140 and 180 mg), C18 (40, 100, 160, 220 and 280 mg) and sodium acetate (0.2, 0.6, 1.0, 1.4 and 1.8 g)) were optimized by the response surface method to obtain the best purification effect. The target compounds were separated on an XBridge-C18 column (100 mm×2.1 mm, 3.5 μm) using acetonitrile-0.1% (v/v) formic acid aqueous solution (containing 1.5 mmol/L ammonium acetate) as the mobile phases by a gradient elution program. Qualitative and quantitative analysis of the target compounds were performed in the full scan and secondary mass spectrometry scan (dd-MS²) modes with positive and negative ionization. The target compounds showed good linear relationships in their respective ranges, with correlation coefficients (R²) greater than 0.99. The limits of detection (LOD, S/N=3) and limits of quantification (LOQ, S/N=10) were in the range of 0.2-1.0 μg/kg and 0.3-3.1 μg/kg, respectively. The average recoveries were 76.4%-116.0% at LOQ, 2-fold LOQ, and 10-fold LOQ levels. The intra-day relative standard deviations (RSDs) were 1.0%-7.9%, and the inter-day RSDs were 1.7%-18.4%. The method is simple, sensitive and accurate, and it is suitable for the determination and quantification of lipid regulators in fish meat.

Key words: ultra performance liquid chromatography (UPLC), quadrupole/electrostatic field orbitrap mass spectrometry (Q/Orbitrap-MS), QuEChERS, lipid regulators, fish meat

WANG Jia, LING Yun, DENG Yamei, ZHANG Yujia, YANG Xusheng, LIU Lixia, LIU Xiaomin, LI Donghui, ZHANG Feng. Determination of 15 lipid regulators in fish meat by QuEChERS-ultra performance liquid chromatography-quadrupole/ electrostatic field orbitrap mass spectrometry[J]. Chinese Journal of Chromatography, 2020, 38(6): 655-662.

Figures/Tables 8

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Compound	t_R/min	Ionization mode	Theoretical mass (m/z)	Measured mass (m/z)	Mass accuracy (10^-6)	Fragment ions (m/z)
t_R: retention time; * quantitative ion.
Pravastatin (普伐他汀)	4.54	ESI^-	423.23583	423.23535^*	0.05	303.15775, 321.16788
Pitavastatin (匹伐他汀)	5.87	ESI^-	420.15666	420.15686^*	0.11	290.13287, 358.15805
Rosuvastatin (罗苏伐他汀)	6.98	ESI^-	480.15661	480.15683^*	0.04	418.15710, 340.18063
Clofibric acid (氯贝酸)	7.60	ESI^-	213.03231	213.03041^*	0.32	85.02734, 126.99335
Bezafibrate (苯扎贝特)	7.77	ESI^-	360.09812	360.09802^*	0.13	274.06216, 154.00383
Ezetimibe (依折麦布)	8.87	ESI⁺	410.15423	410.15420^*	0.07	133.06421, 286.10233
Ciprofibrate (环丙贝特)	9.82	ESI^-	287.02272	287.02246^*	0.14	85.02718, 232.92409
Fluvastatin (氟伐他汀)	9.90	ESI^-	410.17331	410.17343^*	0.04	322.15799, 348.17361
Atorvastatin (阿托伐他汀)	9.98	ESI^-	557.24122	557.24115^*	0.05	278.13361, 397.16815
Cilastatin (西立伐他汀)	11.10	ESI^-	458.23182	458.23169^*	0.15	376.22552, 85.02734
Mevastatin (美伐他汀)	11.90	ESI⁺	391.24379	391.24368^*	0.03	185.13120, 159.11566
Gemfibrozil (吉非贝齐)	12.00	ESI^-	249.14862	249.14842^*	0.12	121.06360
Lovastatin (洛伐他汀)	12.90	ESI⁺	405.26155	405.26141^*	0.09	199.14668, 285.18289
Clinofibrate (克利贝特)	13.50	ESI^-	467.24091	467.24048^*	0.14	367.18869, 267.13693
Fenofibrate (菲诺贝特)	15.20	ESI⁺	361.11751	361.11771^*	0.28	233.03506, 138.99382

Compound	t_R/min	Ionization mode	Theoretical mass (m/z)	Measured mass (m/z)	Mass accuracy (10^-6)	Fragment ions (m/z)
t_R: retention time; * quantitative ion.
Pravastatin (普伐他汀)	4.54	ESI^-	423.23583	423.23535^*	0.05	303.15775, 321.16788
Pitavastatin (匹伐他汀)	5.87	ESI^-	420.15666	420.15686^*	0.11	290.13287, 358.15805
Rosuvastatin (罗苏伐他汀)	6.98	ESI^-	480.15661	480.15683^*	0.04	418.15710, 340.18063
Clofibric acid (氯贝酸)	7.60	ESI^-	213.03231	213.03041^*	0.32	85.02734, 126.99335
Bezafibrate (苯扎贝特)	7.77	ESI^-	360.09812	360.09802^*	0.13	274.06216, 154.00383
Ezetimibe (依折麦布)	8.87	ESI⁺	410.15423	410.15420^*	0.07	133.06421, 286.10233
Ciprofibrate (环丙贝特)	9.82	ESI^-	287.02272	287.02246^*	0.14	85.02718, 232.92409
Fluvastatin (氟伐他汀)	9.90	ESI^-	410.17331	410.17343^*	0.04	322.15799, 348.17361
Atorvastatin (阿托伐他汀)	9.98	ESI^-	557.24122	557.24115^*	0.05	278.13361, 397.16815
Cilastatin (西立伐他汀)	11.10	ESI^-	458.23182	458.23169^*	0.15	376.22552, 85.02734
Mevastatin (美伐他汀)	11.90	ESI⁺	391.24379	391.24368^*	0.03	185.13120, 159.11566
Gemfibrozil (吉非贝齐)	12.00	ESI^-	249.14862	249.14842^*	0.12	121.06360
Lovastatin (洛伐他汀)	12.90	ESI⁺	405.26155	405.26141^*	0.09	199.14668, 285.18289
Clinofibrate (克利贝特)	13.50	ESI^-	467.24091	467.24048^*	0.14	367.18869, 267.13693
Fenofibrate (菲诺贝特)	15.20	ESI⁺	361.11751	361.11771^*	0.28	233.03506, 138.99382

Compound	Liner range/(μg/L)	Linear equation	R²	LOD/(μg/kg)	LOQ/(μg/kg)
Y: peak area; X: mass concentration, μg/L.
Pravastatin	0.2-200	Y=7.77×10⁴X-3.64×10⁵	0.9995	0.2	3.1
Pitavastatin	0.2-200	Y=1.06×10⁵X-2.81×10⁵	0.9965	0.2	0.4
Rosuvastatin	0.2-200	Y=8.84×10⁵X+1.00×10⁷	0.9992	0.2	0.5
Clofibric acid	0.2-200	Y=2.08×10⁵X+5.19×10⁵	0.9936	0.2	0.3
Bezafibrate	0.2-200	Y=5.13×10⁶X-3.75×10⁴	0.9991	0.2	0.4
Ezetimibe	0.2-200	Y=1.01×10⁶X+4.96×10⁶	0.9974	0.2	0.4
Ciprofibrate	0.2-200	Y=1.73×10⁶X+5.7 0×10⁶	0.9988	0.2	2.3
Fluvastatin	1.0-200	Y=1.15×10⁶X-3.77×10⁴	0.9995	1.0	2.1
Atorvastatin	1.0-200	Y=1.13×10⁵X-5.02×10⁵	0.9979	1.0	2.1
Cilastatin	1.0-200	Y=9.58×10⁵X+9.78×10⁶	0.9957	1.0	1.2
Mevastatin	0.5-200	Y=2.06×10⁶X+3.53×10⁶	0.9994	0.5	2.1
Gemfibrozil	0.2-200	Y=2.03×10⁶X+2.55×10⁵	0.9977	0.2	2.1
Lovastatin	1.0-200	Y=9.15×10⁵X+1.56×10⁵	0.9996	1.0	3.0
Clinofibrate	0.2-200	Y=7.07×10⁴X-4.28×10⁵	0.9997	0.2	0.4
Fenofibrate	1.0-200	Y=8.70×10⁵X-1.99×10⁶	0.9978	1.0	2.3

Compound	Liner range/(μg/L)	Linear equation	R²	LOD/(μg/kg)	LOQ/(μg/kg)
Y: peak area; X: mass concentration, μg/L.
Pravastatin	0.2-200	Y=7.77×10⁴X-3.64×10⁵	0.9995	0.2	3.1
Pitavastatin	0.2-200	Y=1.06×10⁵X-2.81×10⁵	0.9965	0.2	0.4
Rosuvastatin	0.2-200	Y=8.84×10⁵X+1.00×10⁷	0.9992	0.2	0.5
Clofibric acid	0.2-200	Y=2.08×10⁵X+5.19×10⁵	0.9936	0.2	0.3
Bezafibrate	0.2-200	Y=5.13×10⁶X-3.75×10⁴	0.9991	0.2	0.4
Ezetimibe	0.2-200	Y=1.01×10⁶X+4.96×10⁶	0.9974	0.2	0.4
Ciprofibrate	0.2-200	Y=1.73×10⁶X+5.7 0×10⁶	0.9988	0.2	2.3
Fluvastatin	1.0-200	Y=1.15×10⁶X-3.77×10⁴	0.9995	1.0	2.1
Atorvastatin	1.0-200	Y=1.13×10⁵X-5.02×10⁵	0.9979	1.0	2.1
Cilastatin	1.0-200	Y=9.58×10⁵X+9.78×10⁶	0.9957	1.0	1.2
Mevastatin	0.5-200	Y=2.06×10⁶X+3.53×10⁶	0.9994	0.5	2.1
Gemfibrozil	0.2-200	Y=2.03×10⁶X+2.55×10⁵	0.9977	0.2	2.1
Lovastatin	1.0-200	Y=9.15×10⁵X+1.56×10⁵	0.9996	1.0	3.0
Clinofibrate	0.2-200	Y=7.07×10⁴X-4.28×10⁵	0.9997	0.2	0.4
Fenofibrate	1.0-200	Y=8.70×10⁵X-1.99×10⁶	0.9978	1.0	2.3

Compound	Spiked level/ (μg/kg)	Average recovery/%	Intra RSD/%	Inter RSD/%
Pravastatin	3.1	78.2	1.6	10.9
	6.2	78.3	1.5	6.6
	31.0	76.4	7.9	16.7
Pitavastatin	0.4	90.2	4.6	9.3
	0.8	92.2	3.7	9.9
	4.0	93.4	4.7	2.0
Rosuvastatin	0.5	100.3	1.5	14.9
	1.0	103.6	1.5	16.8
	5.0	102.7	1.4	10.1
Clofibric acid	0.3	101.2	3.0	18.4
	0.6	100.7	2.9	12.7
	3.0	101.5	1.1	11.3
Bezafibrate	0.4	116.7	2.0	16.4
	0.8	115.6	3.3	17.6
	4.0	112.9	5.6	2.5
Ezetimibe	0.4	95.1	2.0	10.8
	0.8	96.4	4.1	13.1
	4.0	91.0	4.3	4.9
Fenofibrate	2.3	77.9	3.1	9.2
	4.6	79.3	1.4	7.2
	23.0	94.8	1.2	9.7
Fluvastatin	2.1	89.5	1.8	1.7
	4.2	87.6	2.7	5.1
	21.0	90.1	2.3	2.8
Atorvastatin	2.1	77.9	1.0	10.5
	4.2	80.2	3.5	10.6
	21.0	77.6	5.8	2.3
Cilastatin	1.2	103.4	6.2	7.4
	2.4	104.4	2.3	9.1
	12.0	104.0	4.0	6.6
Mevastatin	2.1	90.2	2.3	7.4
	4.2	92.2	1.9	7.6
	21.0	91.4	2.1	2.7
Gemfibrozil	2.1	92.9	5.4	17.8
	4.2	94.2	1.6	11.0
	21.0	93.2	7.9	8.0
Lovastatin	3.0	100.5	4.6	1.9
	6.0	105.9	3.2	3.7
	30.0	107.1	3.2	10.2
Clinofibrate	0.4	91.8	3.5	6.8
	0.8	89.8	3.3	6.1
	4.0	88.9	1.5	5.3
Ciprofibrate	2.3	94.6	1.1	6.9
	4.6	95.5	1.8	11.8
	23.0	82.1	4.2	8.2