Une seule analyse, une seule colonne, et moins de 9 minutes pour plus de 60 antibiotiques de différentes classes

• Excellente séparation des composés et temps d’analyse rapides.
• Polyvalente et applicable à tout type d’analyse de résidus d’antibiotiques — et optimisation possible pour la quantification individuelle de panels de différentes classes :
  • Macrolide, Lincosamide, et Streptogramine (Figure 1)
  • Amphénicol et Tétracycline (Figure 2)
  • Quinolone (Figure 3)
  • Pénicilline, Céphalosporine, et Tétracycline (Figure 4)
  • Sulfonamide (Figure 5) (pour les ionophores, utiliser la colonne Raptor Biphenyl. [Figure 6])

L’utilisation d’antibiotiques sur les animaux élevés pour la consommation humaine est une préoccupation de Santé Publique et de sécurité en raison de la génération potentielle de bactéries résistantes aux médicaments. De nombreux pays dans l’Union Européenne ainsi que le Canada ont interdit l’utilisation d’antibiotiques à des fins non thérapeutiques, et les États-Unis définissent une politique visant à réduire l’utilisation d’antibiotiques médicalement importants pour stimuler la croissance. Afin de réglementer l’usage approprié des antibiotiques vétérinaires, la FDA aux États-Unis a fixé des limites maximales pour les résidus (MRL – “Maximum Residue Limits”) pour différents tissus animaux et produits alimentaires (21 CFR Part 556). Une méthode analytique sensible, efficace et fiable pour différentes classes d’antibiotiques est donc nécessaire pour répondre à ces exigences réglementaires, et la colonne LC Restek Raptor C18 est la colonne idéale pour cela.

LC_FS0506

Peaks

	Peaks	tR (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
1.	Desacetyl cephapirin	0.70	150	382.03	111.92	124.21
2.	Sulfanilamide	0.85	200	172.98	93.07	75.23
3.	Amoxicillin	0.92	100	366.24	349.10	208.07
4.	Cephapirin	1.36	50	424.17	292.08	124.14
5.	Tildipirosin	1.38	200	734.59	561.45	204.15
6.	Desfuroyl ceftiofur cysteine disulfide	1.40	300	549.16	183.02	126.00
7.	Lincomycin	1.50	50	407.32	359.23	389.28
8.	Sulfadiazine	1.57	20	251.18	156.04	92.08
9.	Cefquinome	1.73	200	529.19	134.10	125.12
10.	Ampicillin	1.78	50	350.19	106.07	160.06
11.	Sulfathiazole	1.79	10	256.16	156.03	92.08
12.	Marbofloxacin	1.81	10	363.20	72.11	320.10
13.	Cefalexin	1.82	100	348.10	158.05	174.05
14.	Sulfapyridine	1.86	10	250.13	156.10	92.08
15.	Norfloxacin	1.96	20	320.23	276.20	233.13
16.	Ofloxacin	1.98	10	362.21	318.20	261.15
17.	Sulfamerazine	2.00	20	265.08	156.03	92.08
18.	Cefalonium	2.01	100	459.16	337.03	123.10
19.	Oxytetracycline	2.02	25	461.27	426.15	443.32
20.	Ciprofloxacin	2.04	20	332.18	288.22	245.15
21.	Cefacetrile	2.09	300	362.07	258.08	178.01
22.	Tulathromycin A	2.11	100	806.65	577.42	420.31
23.	Tetracycline	2.21	25	445.28	154.07	427.32
24.	Danofloxacin	2.23	20	358.22	340.16	314.21
25.	Enrofloxacin	2.32	10	360.29	316.22	245.13
26.	Orbifloxacin	2.35	10	396.22	352.17	226.12
27.	Thiamphenicol*	2.38	200	354.16	290.04	184.98
28.	Sulfamethazine	2.39	10	279.23	186.08	124.08
29.	Sulfamethizole	2.52	10	271.17	156.02	108.02
30.	Sulfamethoxypyridazine	2.56	10	281.14	156.03	126.07
31.	Sarafloxacin	2.59	10	386.20	342.20	368.15
32.	Difloxacin	2.65	10	400.23	356.17	299.13
33.	Cefazolin	2.75	100	455.10	323.06	295.09
34.	Spiramycin	2.96	200	843.64	540.36	699.48
35.	Pirlimycin	3.05	20	411.32	363.18	327.21
36.	Chlortetracycline	3.08	25	479.27	154.07	371.06
37.	Sulfachlorpyridazine	3.12	20	285.05	156.03	108.09
38.	Gamithromycin	3.28	100	777.63	619.52	601.45
39.	Sulfadoxine	3.39	10	311.17	156.03	108.09
40.	Sulfamethoxazole	3.46	20	254.18	155.98	147.06
41.	Cefoperazone	3.52	100	646.26	143.07	148.02
42.	Florfenicol*	3.55	200	356.10	336.02	184.98
43.	Sulfaethoxypyridazine	3.56	20	295.17	267.07	156.03
44.	Tilmicosin	3.64	100	869.72	696.50	522.42
45.	Sulfisoxazole	3.76	20	268.14	156.03	113.10
46.	Oxolinic acid	3.94	5	262.10	244.06	215.96
47.	Chloramphenicol*	4.00	200	321.16	151.99	257.04
48.	Ceftiofur	4.11	50	524.14	241.08	125.24
49.	Erythromycin	4.31	25	734.64	576.40	558.38
50.	Sulfadimethoxine	4.36	10	311.17	156.09	108.09
51.	Sulfaquinoxaline	4.42	20	301.18	156.04	108.02
52.	Tylosin**	4.67	100	916.62	772.49	598.36
53.	Penicillin G	5.07	100	335.18	176.07	160.07
54.	Flumequine	5.34	5	262.15	244.11	202.03
55.	Penicillin V	5.56	100	351.10	160.06	114.07
56.	Oxacillin	5.99	100	402.15	160.05	114.06
57.	Virginiamycin M1	6.13	50	526.43	508.31	355.10
58.	Tylvalosin	6.19	50	1042.71	814.46	640.39
59.	Cloxacillin	6.39	100	436.15	277.06	160.05
60.	Nafcillin	6.60	25	415.19	199.09	171.06
61.	Dicloxacillin	6.96	100	470.11	160.05	311.02

*Acquired in negative ion mode

Conditions

Column	Raptor C18 (cat.# 9304A12)
Dimensions:	100 mm x 2.1 mm ID
Particle Size:	2.7 µm
Pore Size:	90 Å
Guard Column:	Raptor C18 EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9304A0252)
Temp.:	35 °C
Standard/Sample
Diluent:	Water
Conc.:	5–300 ng/mL
Inj. Vol.:	2 µL
Mobile Phase
A:	0.1% Formic acid in water
B:	0.1% Formic acid in acetonitrile
	Time (min) Flow (mL/min) %A %B 0.00 0.4 90 10 4.50 0.4 65 35 7.00 0.4 45 55 7.01 0.4 90 10 9.00 0.4 90 10

Detector	MS/MS
Ion Mode:	ESI+/ESI-
Mode:	Scheduled MRM
Instrument	UHPLC
Notes	1. Positive and negative polarity data were collected simultaneously from a single injection. 2. Amphenicol compounds (chloramphenicol, thiamphenicol, and florfenicol) were detected with negative polarity. 3. The MRM was scheduled at +/- 20 to 30 seconds for each analyte. 4. Multiclass antibiotics include penicillin, cephalosporin, tetracycline, sulfonamide, macrolide, lincosamide, streptogramin, amphenicol, and quinolone. **The retention time for Tylosin is noted in the peak list; however, it was not included in the chromatogram. Want even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at www.restek.com/inert.

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Figure 1 : Antibiotiques Macrolide, Lincosamide, et Streptogramine sur Raptor C18 par LC-MS/MS

LC_FS0502

Peaks

	Peaks	tR (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
1.	Tildipirosin	1.01	200	734.59	561.45	204.15
2.	Lincomycin	1.11	50	407.32	359.23	389.28
3.	Tulathromycin A	1.19	100	806.65	577.42	420.31
4.	Spiramycin	1.41	200	843.64	540.36	699.48
5.	Pirlymycin	1.49	20	411.32	363.18	327.21

	Peaks	tR (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
6.	Gamithromycin	1.50	100	777.63	619.52	601.45
7.	Tilmicosin	1.58	100	869.72	696.50	522.42
8.	Erythromycin	1.80	25	734.64	576.40	558.38
9.	Tylosin	1.87	100	916.62	772.49	598.36
10.	Tylvalosin	2.30	50	1042.71	814.46	640.39
11.	Virginiamycin M1	2.45	50	526.43	508.31	355.10

Conditions

Column	Raptor C18 (cat.# 9304A12)
Dimensions:	100 mm x 2.1 mm ID
Particle Size:	2.7 µm
Pore Size:	90 Å
Guard Column:	Raptor C18 EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9304A0252)
Temp.:	40 °C
Standard/Sample
Diluent:	Water
Conc.:	20–200 ng/mL
Inj. Vol.:	2 µL
Mobile Phase
A:	0.1% Formic acid in water
B:	0.1% Formic acid in acetonitrile
	Time (min) Flow (mL/min) %A %B 0.00 0.4 90 10 3.00 0.4 20 80 3.01 0.4 90 10 5.00 0.4 90 10

Detector	MS/MS
Ion Mode:	ESI+
Mode:	MRM
Instrument	UHPLC
Notes	Want even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at www.restek.com/inert.

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Figure 2 : Antibiotiques Amphénicol et Tétracycline sur Raptor C18 par LC-MS/MS

LC_FS0504

Peaks

	Peaks	tR (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
1.	Oxytetracycline	1.28	25	461.27	426.15	443.32
2.	Tetracycline	1.34	25	445.28	154.07	427.32
3.	Thiamphenicol*	1.48	200	354.16	290.04	184.98
4.	Chlortetracycline	1.56	25	479.27	154.07	371.06
5.	Florfenicol*	1.86	200	356.10	336.02	184.98
6.	Chloramphenicol*	1.95	200	321.16	151.99	257.04

*Acquired in negative ion mode.

Conditions

Column	Raptor C18 (cat.# 9304A12)
Dimensions:	100 mm x 2.1 mm ID
Particle Size:	2.7 µm
Pore Size:	90 Å
Guard Column:	Raptor C18 EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9304A0252)
Temp.:	40 °C
Standard/Sample
Diluent:	Water
Conc.:	25–200 ng/mL
Inj. Vol.:	2 µL
Mobile Phase
A:	0.1% Formic acid in water
B:	0.1% Formic acid in acetonitrile
	Time (min) Flow (mL/min) %A %B 0.00 0.4 90 10 3.00 0.4 20 80 3.01 0.4 90 10 5.00 0.4 90 10

Detector	MS/MS
Ion Mode:	ESI+/ESI-
Mode:	MRM
Instrument	UHPLC
Notes	Tetracyclines and amphenicols were analyzed with ESI+ and ESI- mode, respectively. Want even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at www.restek.com/inert.

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Figure 3 : Antibiotiques Quinolone sur Raptor C18 par LC-MS/MS

LC_FS0505

Peaks

	Peaks	tR (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
1.	Marbofloxacin	1.28	10	363.20	72.11	320.10
2.	Norfloxacin	1.32	20	320.23	276.20	233.13
3.	Ofloxacin	1.32	10	362.21	318.20	261.15
4.	Ciprofloxacin	1.35	20	332.18	288.22	245.15
5.	Danofloxacin	1.40	20	358.22	340.16	314.21

	Peaks	tR (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
6.	Enrofloxacin	1.44	10	360.29	316.22	245.13
7.	Orbifloxacin	1.47	10	396.22	352.17	226.12
8.	Sarafloxacin	1.55	10	386.20	342.20	368.15
9.	Difloxacin	1.57	10	400.23	356.17	299.13
10.	Oxolinic acid	2.23	5	262.10	244.06	215.96
11.	Flumequine	2.78	5	262.15	244.11	202.03

Conditions

Column	Raptor C18 (cat.# 9304A12)
Dimensions:	100 mm x 2.1 mm ID
Particle Size:	2.7 µm
Pore Size:	90 Å
Guard Column:	Raptor C18 EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9304A0252)
Temp.:	40 °C
Standard/Sample
Diluent:	Water
Conc.:	5–20 ng/mL
Inj. Vol.:	2 µL
Mobile Phase
A:	0.1% Formic acid in water
B:	0.1% Formic acid in acetonitrile
	Time (min) Flow (mL/min) %A %B 0.00 0.4 90 10 3.00 0.4 40 60 3.01 0.4 90 10 5.00 0.4 90 10

Detector	MS/MS
Ion Mode:	ESI+
Mode:	MRM
Instrument	UHPLC
Notes	Want even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at www.restek.com/inert.

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Figure 4 : Antibiotiques Pénicilline, Céphalosporine et Tétracycline sur Raptor C18 par LC-MS/MS

LC_FS0500

Peaks

	Peaks	tR (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
1.	Desacetyl cephapirin	0.68	150	382.03	111.92	124.21
2.	Amoxicillin	0.89	100	366.24	349.10	208.07
3.	Cephapirin	1.05	50	424.17	292.08	124.14
4.	Desfuroyl ceftiofur cysteine disulfide	1.06	300	549.16	183.02	126.00
5.	Cefquinome	1.16	200	529.19	134.10	125.12
6.	Ampicillin	1.18	50	350.19	106.07	160.06
7.	Cefalexin	1.19	100	348.10	158.05	174.05
8.	Oxytetracycline	1.26	50	461.27	426.15	443.32
9.	Cefalonium	1.29	100	459.16	337.03	123.10
10.	Tetracycline	1.32	50	445.28	154.07	427.32

	Peaks	tR (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
11.	Cefacetrile	1.37	300	362.07	258.08	178.01
12.	Cefazolin	1.50	100	455.10	323.06	295.09
13.	Chlortetracycline	1.54	50	479.27	154.07	371.06
14.	Cefoperazone	1.69	100	646.26	143.07	148.02
15.	Ceftiofur	1.85	50	524.14	241.08	125.24
16.	Penicillin G	2.18	100	335.18	176.07	160.07
17.	Penicillin V	2.33	100	351.10	160.06	114.07
18.	Oxacillin	2.44	100	402.15	160.05	114.06
19.	Cloxacillin	2.56	100	436.15	277.06	160.05
20.	Nafcillin	2.63	25	415.19	199.09	171.06
21.	Dicloxacillin	2.76	100	470.11	160.05	311.02

Conditions

Column	Raptor C18 (cat.# 9304A12)
Dimensions:	100 mm x 2.1 mm ID
Particle Size:	2.7 µm
Pore Size:	90 Å
Guard Column:	Raptor C18 EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9304A0252)
Temp.:	40 °C
Standard/Sample
Diluent:	Water
Conc.:	25–300 ng/mL
Inj. Vol.:	2 µL
Mobile Phase
A:	0.1% Formic acid in water
B:	0.1% Formic acid in acetonitrile
	Time (min) Flow (mL/min) %A %B 0.00 0.4 90 10 3.00 0.4 20 80 3.01 0.4 90 10 5.00 0.4 90 10

Detector	MS/MS
Ion Mode:	ESI+
Mode:	MRM
Instrument	UHPLC
Notes	Want even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at www.restek.com/inert.

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Figure 5 : Antibiotiques Sulfonamide sur Raptor C18 par LC-MS/MS

LC_FS0501

Peaks

	Peaks	tR (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
1.	Sulfanilamide	0.83	200	172.98	93.07	75.23
2.	Sulfadiazine	1.45	20	251.18	156.04	92.08
3.	Sulfathiazole	1.60	10	256.16	156.03	92.08
4.	Sulfapyridine	1.67	10	250.13	156.10	92.08
5.	Sulfamerazine	1.79	20	265.08	156.03	92.08
6.	Sulfamethazine	2.07	10	279.23	186.08	124.08
7.	Sulfamethizole	2.11	10	271.17	156.02	108.02

	Peaks	tR (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
8.	Sulfamethoxypyridazine	2.16	10	281.14	156.03	126.07
9.	Sulfachlorpyridazine	2.55	20	285.05	156.03	108.09
10.	Sulfadoxine	2.75	10	311.17	156.03	108.09
11.	Sulfamethoxazole	2.78	20	254.18	155.98	147.06
12.	Sulfaethoxypyridazine	2.84	20	295.17	267.07	156.03
13.	Sulfisoxazole	2.98	20	268.14	156.03	113.10
14.	Sulfadimethoxine	3.37	10	311.17	156.09	108.09
15.	Sulfaquinoxaline	3.40	20	301.18	156.04	108.02

Conditions

Column	Raptor C18 (cat.# 9304A12)
Dimensions:	100 mm x 2.1 mm ID
Particle Size:	2.7 µm
Pore Size:	90 Å
Guard Column:	Raptor C18 EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9304A0252)
Temp.:	40 °C
Standard/Sample
Diluent:	Water
Conc.:	10–200 ng/mL
Inj. Vol.:	2 µL
Mobile Phase
A:	0.1% Formic acid in water
B:	0.1% Formic acid in acetonitrile
	Time (min) Flow (mL/min) %A %B 0.00 0.4 90 10 3.50 0.4 60 40 3.51 0.4 90 10 5.00 0.4 90 10

Detector	MS/MS
Ion Mode:	ESI+
Mode:	MRM
Instrument	UHPLC

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Figure 6 : Antibiotiques Ionophores sur Raptor Biphenyl par LC-MS/MS

LC_FS0503

Peaks

	Peaks	tR (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
1.	Lasalocid A	1.92	100	613.42	377.28	595.40
2.	Monensin	2.12	100	693.50	675.44	461.30
3.	Salinomycin	2.19	100	773.57	431.24	531.39
4.	Maduramicin	2.30	100	939.65	877.58	473.31
5.	Narasin	2.58	100	787.59	431.27	531.35

Conditions

Column	Raptor Biphenyl (cat.# 9309A12)
Dimensions:	100 mm x 2.1 mm ID
Particle Size:	2.7 µm
Pore Size:	90 Å
Guard Column:	Raptor Biphenyl EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9309A0252)
Temp.:	40 °C
Standard/Sample
Diluent:	Water:methanol (10:90)
Conc.:	100 ng/mL
Inj. Vol.:	2 µL
Mobile Phase
A:	0.5% Formic acid in water
B:	0.5% Formic acid in acetonitrile
	Time (min) Flow (mL/min) %A %B 0.00 0.5 35 65 3.00 0.5 25 75 3.01 0.5 35 65 5.00 0.5 35 65

Detector	MS/MS
Ion Mode:	ESI+
Mode:	MRM
Instrument	UHPLC
Notes	Want even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at www.restek.com/inert.

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FSSS2276-FR

Version: A