Study for the determination of high-risk quinolone antibiotics residues and its matrix effect in cultured aquatic products by UPLC-MS/MS based on response surface analysis

doi:10.3969/j.issn.1004-1524.20240352

Acta Agriculturae Zhejiangensis ›› 2025, Vol. 37 ›› Issue (3): 689-700.DOI: 10.3969/j.issn.1004-1524.20240352

• Quality and Safety of Agricultural Products • Previous Articles Next Articles

Study for the determination of high-risk quinolone antibiotics residues and its matrix effect in cultured aquatic products by UPLC-MS/MS based on response surface analysis

XU Huibin¹^,²^,³(), ZHU Jie¹^,²^,³^,^*(), ZHOU Chaosheng¹^,²^,³, HU yuan¹^,²^,³, LU Rongmao¹^,²^,³

1. Zhejiang Mariculture Research Institute, Wenzhou 325005, Zhejiang, China
2. Zhejiang Key Laboratory of Exploitation and Preservation of Coastal Bio-Resource, Wenzhou 325005, Zhejiang, China
3. Zhejiang Key Laboratory of Coastal Biological Germplasm Resources Conservation and Utilization, Wenzhou 325005, Zhejiang, China

Received:2024-04-16 Online:2025-03-25 Published:2025-04-02

Abstract

Abstract:

In order to establish an general, efficient and accurate residue analysis method for high risk quinolone antibiotics in cultured aquatic products, such as Larimichthys crocea, Carassius auratus, Pelodiscus sinensis, etc, all the samples were extracted using ultrasonic and vibrating extraction coupling, and then cleaned by liquid-liquid extraction and determined by UPLC-MS/MS. The optimal pretreatment conditions were determined by combining hydrochloric acid concentration in extraction reagent, oscillation time, ultrasonic time, nitrogen blowing pressure, temperature with Box-Behnken response surface analysis. The matrix effects of the target objects were evaluated in this study, and the compensation method of matrix effects in Larimichthys crocea, Carassius auratus, Pelodiscus sinensis, Penaeus vannemei, Scylla serrata and its gonad, and Sinonovacula constricta was discussed by isotope internal standard and matrix matching curve. The results showed that the average recovery rate of 6 target substances was 82.20%-118.00%. The linear correlation coefficients were above 0.997 in spiked range of 0.2-20 ng·mL^-1. The limits of detection and limits of quantitation were 0.5 μg·kg^-1 and 1.0 μg·kg^-1, respectively. It showed that the method met the requirements for the analysis of antibiotic residues in cultured aquatic products. To varying degrees, the matrix of Penaeus vannamei, Scylla serrata gonad and Larimichthys crocea inhibited the 6 target substances. The matrix matching curve and internal standard method could compensate the matrix effect effectively.

Key words: response surface, UPLC-MS/MS, cultured aquatic product, high risk, quinolone, matrix effects evaluation

CLC Number:

S859.84

XU Huibin, ZHU Jie, ZHOU Chaosheng, HU yuan, LU Rongmao. Study for the determination of high-risk quinolone antibiotics residues and its matrix effect in cultured aquatic products by UPLC-MS/MS based on response surface analysis[J]. Acta Agriculturae Zhejiangensis, 2025, 37(3): 689-700.

Figures/Tables 14

References 14

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时间t/min	A/%	B/%
0.30	5.0	95.0
5.40	50.0	50.0
5.80	50.0	50.0
6.00	5.0	95.0

时间t/min	A/%	B/%
0.30	5.0	95.0
5.40	50.0	50.0
5.80	50.0	50.0
6.00	5.0	95.0

待测化合物 Target	母离子 Parents	子离子 Daughter	锥孔电压 Cone/V	碰撞能量 Collision/eV
氧氟沙星	362	318^*	32	26
Ofloxacin		261		18
洛美沙星	352	265^*	32	22
Lomefloxacin		334		19
培氟沙星	334	290^*	32	24
Pefloxacin		233		17
诺氟沙星	320	302^*	32	22
Norfloxacin		233		20
恩诺沙星	360	316^*	32	24
Enrofloxacin		245		18
环丙沙星	332	314^*	32	35
Ciprofloxacin		231		22

待测化合物 Target	母离子 Parents	子离子 Daughter	锥孔电压 Cone/V	碰撞能量 Collision/eV
氧氟沙星	362	318^*	32	26
Ofloxacin		261		18
洛美沙星	352	265^*	32	22
Lomefloxacin		334		19
培氟沙星	334	290^*	32	24
Pefloxacin		233		17
诺氟沙星	320	302^*	32	22
Norfloxacin		233		20
恩诺沙星	360	316^*	32	24
Enrofloxacin		245		18
环丙沙星	332	314^*	32	35
Ciprofloxacin		231		22

处理Treatment	OFL	LOM	NOR	PEF	ENR	CIP
乙腈ACN	55.00±7.35 b	41.33±1.41 c	21.17±2.09 c	65.00±17.44 c	74.57±20.55 b	27.27±1.62 c
2% HCl(1+1)-ACN	97.70±4.00 a	90.70±4.36 a	85.80±3.70 a	100.97±5.03 a	106.10±5.03 a	75.20±4.26 a
2% HCOOH-ACN	68.93±13.70 b	66.50±12.84 b	55.63±12.16 b	66.33±14.31 b	55.33±14.30 b	48.73±8.80 b

Study for the determination of high-risk quinolone antibiotics residues and its matrix effect in cultured aquatic products by UPLC-MS/MS based on response surface analysis

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Figures/Tables 14

References 14

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水平 Level	因素Factor
水平 Level	氮吹温度 Nitrogen blowing temperature/℃	盐酸浓度 Hydrochloric acid concentration/%	振荡时间 Oscillation time/min	超声时间 Ultrasound time/min
-1	40	0	0	0
0	50	1	5	15
1	70	2	10	30

轮次 Run	氮吹温度 Nitrogen blowing temperature/℃	盐酸浓度 Hydrochloric acid concentration/%	振荡时间 Oscillation time/min	超声时间 Ultrasound time/min	回收率 Recovery rate/%
1	-1	0	0	1	55.3
2	0	-1	0	1	94.8
3	-1	0	1	0	94.7
4	0	1	0	1	85.9
5	0	0	-1	1	88.9
6	1	0	-1	0	85.9
7	0	0	0	0	70.4
8	-1	-1	0	0	50.1
9	1	0	0	1	72.0
10	0	-1	-1	0	66.0
11	0	-1	1	0	112.8
12	1	-1	0	0	95.2
13	0	1	1	0	105.3
14	1	1	0	0	95.7
15	-1	0	0	-1	95.3
16	0	0	1	-1	104.0
17	0	1	-1	0	102.9
18	0	0	0	0	100.9
19	1	0	0	-1	94.1
20	0	0	1	1	86.3
21	0	-1	0	-1	90.7
22	0	0	0	0	90.7
23	0	1	0	-1	94.2
24	-1	1	0	0	46.1
25	0	0	0	0	83.1
26	0	0	0	0	75.9
27	0	0	-1	-1	78.5
28	-1	0	-1	0	73.8
29	1	0	1	0	80.3

方差来源 Source	平方和 SS	自由度 dF	均方 MS	F值 F value	P值(Prob >F) P value	显著性 Significance
模型	6 970.19	14	497.87	12.86	<0.000 1	**
A	383.70	1	383.70	9.910	0.007 1	**
B	2 353.71	1	2 353.71	60.79	<0.000 1	**
C	51.28	1	51.28	1.320	0.269 1	不显著Not significant
D	6.20	1	6.20	0.160	0.695 2	不显著Not significant
A²	1 337.37	1	1 337.37	34.540	<0.000 1	**
B²	2 930.16	1	2 930.16	75.680	<0.000 1	**
C²	0.58	1	0.58	0.015	0.904 0	不显著Not significant
D²	28.09	1	28.09	0.730	0.408 7	不显著Not significant
AB	3.24	1	3.24	0.084	0.776 6	不显著Not significant
AC	2.80	1	2.80	0.072	0.792 1	不显著Not significant
AD	32.11	1	32.11	0.830	0.377 9	不显著Not significant
BC	38.10	1	38.10	0.980	0.338 1	不显著Not significant
BD	121.18	1	121.18	3.130	0.098 6	不显著Not significant
CD	29.31	1	29.31	0.760	0.398 9	不显著Not significant
残项Residual	542.07	14	38.72
失拟项Lack of fit	451.23	10	45.12	1.990	0.265 4	不显著Not significant
纯误差Pure error	90.84	4	22.71
总和Sum	7 512.27	28	497.87	12.860

标号 No.	基质类型 Varieties	氧氟沙星 OFL	洛美沙星 LOM	诺氟沙星 NOR	培氟沙星 PEF	恩诺沙星 ENR	环丙沙星 CIP
1	缢蛏Sinonovacula constricta	0.99	0.99	0.93	1.15	1.09	0.92
2	中华鳖Trionyx sinensis	0.99	0.97	0.96	1.13	1.04	0.94
3	石蛙Quasipaa spinosa	0.93	0.94	0.93	1.06	1.00	0.90
4	南美白对虾1 No. 1 of Litopenaeus Vannamei	0.59	0.66	0.71	0.67	0.60	0.71
	南美白对虾2 No.2 of Litopenaeus Vannamei	0.75	0.69	0.65	0.78	0.75	0.57
5	大黄鱼Larimichthys crocea	0.79	0.80	0.79	0.83	0.83	0.78
6	鲫鱼Carassius auratus	0.88	0.86	0.85	1.01	0.94	0.84
7	青蟹Scylla serrata	1.03	0.96	0.84	1.06	1.02	0.78
8	青蟹性腺Gonad of Scylla serrata	0.71	0.67	0.64	0.82	0.73	0.58

目标分析物 Target	加标量 Addition amount/ (μg·kg^-1)	回收率Recovery rate/%
目标分析物 Target	加标量 Addition amount/ (μg·kg^-1)	溶剂曲线计算 Solvent curve	基质曲线矫正 Matrix curve	内标矫正 Internal standard correction
氧氟沙星OFL	5.0	43.9	78.0	78.9
洛美沙星LOM	5.0	39.2	75.6	80.2
诺氟沙星NOR	5.0	42.7	68.1	84.8
培氟沙星PEF	5.0	59.5	73.6	83.4
恩诺沙星ENR	5.0	47.8	64.9	85.1
环丙沙星CIP	5.0	41.0	57.1	87.1

水产品 Aquatic products	加标量 Addition amount/ (μg·kg^-1)	氧氟沙星OLF		洛美沙星LOM		诺氟沙星NOR		培氟沙星PEF		恩诺沙星ENR		环丙沙星CIP
水产品 Aquatic products	加标量 Addition amount/ (μg·kg^-1)	回收率 Recovery rate	偏差 RSD	回收率 Recovery rate	偏差 RSD	回收率 Recovery rate	偏差 RSD	回收率 Recovery rate	偏差 RSD	回收率 Recovery rate	偏差 RSD	回收率 Recovery rate	偏差 RSD
大黄鱼	1	89.17	3.38	94.67	5.08	110.00	7.32	112.00	6.76	104.50	4.56	88.33	5.58
Larimichthys	5	84.77	2.55	94.37	6.01	93.73	2.02	86.10	1.57	95.50	4.77	89.83	1.65
crocea	10	84.73	9.13	97.47	9.47	100.27	5.58	93.97	3.18	92.23	1.07	83.17	2.75
	100	94.02	1.47	90.67	2.25	94.24	4.08	99.59	0.20	96.71	2.75	106.39	2.22
中华鳖	1	117.00	4.27	113.50	6.06	101.17	4.95	111.00	2.25	118.00	1.12	95.33	7.68
Trionyx	5	90.83	3.26	102.30	5.65	104.63	3.45	92.97	3.25	91.57	3.12	90.57	2.07
sinensis	10	92.90	1.56	105.43	2.35	105.93	2.10	95.30	2.38	90.50	0.92	93.37	0.90
	100	93.91	0.87	91.03	1.56	99.12	2.55	100.49	3.09	96.39	1.38	102.62	0.76
石蛙	1	87.83	7.21	83.83	3.60	98.33	9.54	98.17	2.06	104.33	2.73	100.00	3.50
Quasipaa	5	93.30	1.04	82.60	3.80	96.00	2.86	97.97	3.37	88.13	1.54	84.67	1.85
spinosa	10	92.53	1.45	104.10	1.21	107.37	2.32	95.93	1.29	82.20	1.08	87.63	2.75
	100	96.04	1.95	90.35	1.00	103.43	2.14	101.88	1.11	101.42	0.79	114.65	1.47

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