One Analysis, One Column, Less than 9 Minutes for Over 60 Multiclass Antibiotics

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Featured Application: Multiclass Veterinary Antibiotics on Raptor C8 by LC- One Analysis, One Column, Less than 9 Minutes for Over 0 Multiclass Antibiotics Highly efficient peak separation and fast analysis times. Versatility and global applicability for antibiotic residue analysis capable of individual class panel optimization for quantitation: - Macrolide, Lincosamide, and Streptogramin (Figure ) - Amphenicol and Tetracycline (Figure ) - Quinolone (Figure ) - Penicillin, Cephalosporin, and Tetracycline (Figure 4) - Sulfonamide (Figure 5) (For Ionophore, use on Raptor Biphenyl. [Figure ]) The use of antibiotics on food-producing animals is a public health and safety concern due to the potential of generating drug-resistant bacteria. Many countries in the European Union and Canada have banned the use of antibiotics for nontherapeutic purposes, and the United States is implementing a policy to reduce the use of medically important antibiotics for growth promotion. To regulate the proper use of veterinary antibiotics, the U.S. FDA has set maximum residue limits (MRL) for a variety of animal tissue and food products ( CFR Part 55). A sensitive, efficient, and reliable analytical method for different classes of antibiotics is necessary to meet this regulation, and the Raptor C8 LC column is the ideal choice.. Desacetyl cephapirin 0.70 50 8.0.9 4.. Sulfanilamide 0.85 00 7.98 9.07 75.. Amoxicillin 0.9 00.4 49.0 08.07 4. Cephapirin. 50 44.7 9.08 4.4 5. Tildipirosin.8 00 74.59 5.45 04.5. Desfuroyl ceftiofur cysteine disulfide.40 00 549. 8.0.00 7. Lincomycin.50 50 407. 59. 89.8 8. Sulfadiazine.57 0 5.8 5.04 9.08 9. Cefquinome.7 00 59.9 4.0 5. 0. Ampicillin.78 50 50.9 0.07 0.0. Sulfathiazole.79 0 5. 5.0 9.08. Marbofloxacin.8 0.0 7. 0.0. Cefalexin.8 00 48.0 58.05 74.05 4. Sulfapyridine.8 0 50. 5.0 9.08 5. Norfloxacin.9 0 0. 7.0.. Ofloxacin.98 0. 8.0.5 7. Sulfamerazine.00 0 5.08 5.0 9.08 8. Cefalonium.0 00 459. 7.0.0 9. Oxytetracycline.0 5 4.7 4.5 44. 0. Ciprofloxacin.04 0.8 88. 45.5. Cefacetrile.09 00.07 58.08 78.0. Tulathromycin A. 00 80.5 577.4 40.. Tetracycline. 5 445.8 54.07 47. 4. Danofloxacin. 0 58. 40. 4. 5. Enrofloxacin. 0 0.9. 45.. Orbifloxacin.5 0 9. 5.7. 7. Thiamphenicol*.8 00 54. 90.04 84.98 8. Sulfamethazine.9 0 79. 8.08 4.08 9. Sulfamethizole.5 0 7.7 5.0 08.0 0. Sulfamethoxypyridazine.5 0 8.4 5.0.07. Sarafloxacin.59 0 8.0 4.0 8.5. Difloxacin.5 0 400. 5.7 99.. Cefazolin.75 00 455.0.0 95.09 4. Spiramycin.9 00 84.4 540. 99.48 5. Pirlimycin.05 0 4..8 7.. Chlortetracycline.08 5 479.7 54.07 7.0 7. Sulfachlorpyridazine. 0 85.05 5.0 08.09 8. Gamithromycin.8 00 777. 9.5 0.45 9. Sulfadoxine.9 0.7 5.0 08.09 40. Sulfamethoxazole.4 0 54.8 55.98 47.0 4. Cefoperazone.5 00 4. 4.07 48.0 4. Florfenicol*.55 00 5.0.0 84.98 4. Sulfaethoxypyridazine.5 0 95.7 7.07 5.0 44. Tilmicosin.4 00 89.7 9.50 5.4 45. Sulfisoxazole.7 0 8.4 5.0.0 4. Oxolinic acid.94 5.0 44.0 5.9 47. Chloramphenicol* 4.00 00. 5.99 57.04 48. Ceftiofur 4. 50 54.4 4.08 5.4 49. Erythromycin 4. 5 74.4 57.40 558.8 50. Sulfadimethoxine 4. 0.7 5.09 08.09 5. Sulfaquinoxaline 4.4 0 0.8 5.04 08.0 5. Tylosin** 4.7 00 9. 77.49 598. 5. Penicillin G 5.07 00 5.8 7.07 0.07 54. Flumequine 5.4 5.5 44. 0.0 55. Penicillin V 5.5 00 5.0 0.0 4.07 5. Oxacillin 5.99 00 40.5 0.05 4.0 57. Virginiamycin M. 50 5.4 508. 55.0 58. Tylvalosin.9 50 04.7 84.4 40.9 59. Cloxacillin.9 00 4.5 77.0 0.05 0. Nafcillin.0 5 45.9 99.09 7.0. Dicloxacillin.9 00 470. 0.05.0 *Acquired in negative ion mode See chromatogram and conditions on page www.restek.com

0.00 0.50.00.50.00.50.00.50 4.00 4.50 5.00 5.50.00.50 7.00 7.50 8.00 LC_FS050 Column Raptor C8 (cat.# 904A) (cat.# 904A05) Temp.: 5 C : 5 00 ng/ml Notes 4.50 0.4 5 5 7.00 0.4 45 55 7.0 0.4 90 0 9.00 0.4 90 0 /ESI- Scheduled. Positive and negative polarity data were collected simultaneously from a single injection.. Amphenicol compounds (chloramphenicol, thiamphenicol, and florfenicol) were detected with negative polarity.. The was scheduled at +/- 0 to 0 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. www.restek.com

Figure : Macrolide, Lincosamide, and Streptogramin Antibiotics on Raptor C8 by LC- 8 5 9 7 0 4 0.00 0.0 0.40 0.0 0.80.00.0.40.0.80.00.0.40.0.80.00.0.40 LC_FS050. Tildipirosin.0 00 74.59 5.45 04.5. Lincomycin. 50 407. 59. 89.8. Tulathromycin A.9 00 80.5 577.4 40. 4. Spiramycin.4 00 84.4 540. 99.48 5. Pirlymycin.49 0 4..8 7.. Gamithromycin.50 00 777. 9.5 0.45 7. Tilmicosin.58 00 89.7 9.50 5.4 8. Erythromycin.80 5 74.4 57.40 558.8 9. Tylosin.87 00 9. 77.49 598. 0. Tylvalosin.0 50 04.7 84.4 40.9. Virginiamycin M.45 50 5.4 508. 55.0 Column Raptor C8 (cat.# 904A) (cat.# 904A05) : 0 00 ng/ml.00 0.4 0 80.0 0.4 90 0 www.restek.com

Figure : Amphenicol and Tetracycline Antibiotics on Raptor C8 by LC- 5 4 0.00 0.0 0.40 0.0 0.80.00.0.40.0.80.00.0.40.0.80.00 LC_FS0504. Oxytetracycline.8 5 4.7 4.5 44.. Tetracycline.4 5 445.8 54.07 47.. Thiamphenicol*.48 00 54. 90.04 84.98 4. Chlortetracycline.5 5 479.7 54.07 7.0 5. Florfenicol*.8 00 5.0.0 84.98. Chloramphenicol*.95 00. 5.99 57.04 *Acquired in negative ion mode. Column Raptor C8 (cat.# 904A) (cat.# 904A05) : 5 00 ng/ml Notes.00 0.4 0 80.0 0.4 90 0 /ESI- Tetracyclines and amphenicols were analyzed with and ESI- mode, respectively. 4 www.restek.com

Figure : Quinolone Antibiotics on Raptor C8 by LC- 7 0 9 4 8 5 0.00 0.5 0.50 0.75.00.5.50.75.00.5.50.75.00.5.50.75 4.00 LC_FS0505. Marbofloxacin.8 0.0 7. 0.0. Norfloxacin. 0 0. 7.0.. Ofloxacin. 0. 8.0.5 4. Ciprofloxacin.5 0.8 88. 45.5 5. Danofloxacin.40 0 58. 40. 4.. Enrofloxacin.44 0 0.9. 45. 7. Orbifloxacin.47 0 9. 5.7. 8. Sarafloxacin.55 0 8.0 4.0 8.5 9. Difloxacin.57 0 400. 5.7 99. 0. Oxolinic acid. 5.0 44.0 5.9. Flumequine.78 5.5 44. 0.0 Column Raptor C8 (cat.# 904A) (cat.# 904A05) : 5 0 ng/ml.00 0.4 40 0.0 0.4 90 0 www.restek.com 5

Figure 4: Penicillin, Cephalosporin, and Tetracycline Antibiotics on Raptor C8 by LC- 0 8 5 9 7 8 9 0 4 4 5 7 0.00 0.0 0.40 0.0 0.80.00.0.40.0.80.00.0.40.0.80.00.0.40 LC_FS0500. Desacetyl cephapirin 0.8 50 8.0.9 4.. Amoxicillin 0.89 00.4 49.0 08.07. Cephapirin.05 50 44.7 9.08 4.4 4. Desfuroyl ceftiofur.0 00 549. 8.0.00 cysteine disulfide 5. Cefquinome. 00 59.9 4.0 5.. Ampicillin.8 50 50.9 0.07 0.0 7. Cefalexin.9 00 48.0 58.05 74.05 8. Oxytetracycline. 50 4.7 4.5 44. 9. Cefalonium.9 00 459. 7.0.0 0. Tetracycline. 50 445.8 54.07 47.. Cefacetrile.7 00.07 58.08 78.0. Cefazolin.50 00 455.0.0 95.09. Chlortetracycline.54 50 479.7 54.07 7.0 4. Cefoperazone.9 00 4. 4.07 48.0 5. Ceftiofur.85 50 54.4 4.08 5.4. Penicillin G.8 00 5.8 7.07 0.07 7. Penicillin V. 00 5.0 0.0 4.07 8. Oxacillin.44 00 40.5 0.05 4.0 9. Cloxacillin.5 00 4.5 77.0 0.05 0. Nafcillin. 5 45.9 99.09 7.0. Dicloxacillin.7 00 470. 0.05.0 Column Raptor C8 (cat.# 904A) (cat.# 904A05) : 5 00 ng/ml.00 0.4 0 80.0 0.4 90 0 www.restek.com

Figure 5: Sulfonamide Antibiotics on Raptor C8 by LC- 4 4 7 8 9 5 0 5 0.00 0.5 0.50 0.75.00.5.50.75.00.5.50.75.00.5.50.75 4.00 LC_FS050. Sulfanilamide 0.8 00 7.98 9.07 75.. Sulfadiazine.45 0 5.8 5.04 9.08. Sulfathiazole.0 0 5. 5.0 9.08 4. Sulfapyridine.7 0 50. 5.0 9.08 5. Sulfamerazine.79 0 5.08 5.0 9.08. Sulfamethazine.07 0 79. 8.08 4.08 7. Sulfamethizole. 0 7.7 5.0 08.0 8. Sulfamethoxypyridazine. 0 8.4 5.0.07 9. Sulfachlorpyridazine.55 0 85.05 5.0 08.09 0. Sulfadoxine.75 0.7 5.0 08.09. Sulfamethoxazole.78 0 54.8 55.98 47.0. Sulfaethoxypyridazine.84 0 95.7 7.07 5.0. Sulfisoxazole.98 0 8.4 5.0.0 4. Sulfadimethoxine.7 0.7 5.09 08.09 5. Sulfaquinoxaline.40 0 0.8 5.04 08.0 Column Raptor C8 (cat.# 904A) (cat.# 904A05) : 0 00 ng/ml.50 0.4 0 40.5 0.4 90 0 www.restek.com 7

Figure : Ionophore Antibiotics on Raptor Biphenyl by LC- 4 5 0.00 0.5 0.50 0.75.00.5.50.75.00.5.50.75.00.5.50.75 4.00 LC_FS050. Lasalocid A.9 00.4 77.8 595.40. Monensin. 00 9.50 75.44 4.0. Salinomycin.9 00 77.57 4.4 5.9 4. Maduramicin.0 00 99.5 877.58 47. 5. Narasin.58 00 787.59 4.7 5.5 Column Raptor Biphenyl (cat.# 909A) Guard Column: Raptor Biphenyl EXP guard column cartridge 5 mm,. mm ID,.7 µm (cat.# 909A05) :methanol (0:90) : 00 ng/ml A: 0.5% Formic acid in water B: 0.5% Formic acid in acetonitrile 0.00 0.5 5 5.00 0.5 5 75.0 0.5 5 5 5.00 0.5 5 5 Questions about this or any other Restek product? Contact us or your local Restek representative (www.restek.com/contact-us). Restek patents and trademarks are the property of Restek Corporation. (See www.restek.com/patents-trademarks for full list.) Other trademarks in Restek literature or on its website are the property of their respective owners. Restek registered trademarks are registered in the U.S. and may also be registered in other countries. 08 Restek Corporation. All rights reserved. Printed in the U.S.A. www.restek.com Lit. Cat.# FSSS7A-UNV