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Research Article


Validation method for determining the concentration of norfloxacin-tylosin combination in broiler chicken tissue using high-performance liquid chromatography

Cahyo Wibisono, Agustina Dwi Wijayanti, Dyah Ayu Widiasih, Ida Tjahajati, Ika Nindya Irianti, Kanya Gabriella Sasri Pamudya, Ridha Avicena Ila Salsabila, Luh Made Sudimartini.


Abstract
Background:
The combination of norfloxacin and tylosin is an alternative to prevent antibiotic residue and resistance. Measurement of the levels of the combination of norfloxacin-tylosin in tissue using high-performance liquid chromatography (HPLC) to obtain parameters for further research, such as pharmacokinetics and residue analysis. When broiler chickens are given norfloxacin-tylosin antibiotics, they make antibiotics more effective against bacterial illnesses. These results indicate that the right mix of antibiotics can be used as a single medicine to achieve synergistic effects. Also, reach the SDGs for no hunger, good health, and well-being.

Aim:
This study aimed to ascertain the correct validation method for the combination antibiotic norfloxacin-tylosin in broiler chicken tissues, specifically the liver, kidney, and muscle.

Methods:
HPLC consistently detected norfloxacin-tylosin in broiler chicken tissue. Mobile phase methanol/acetonitrile was 50:50 (v/v). The chromatographic equipment utilized a 250 mm Shimpack ODS C18 column with a four μm diameter. Solution detection happened at 1 mL/min flow rate, 30°C oven temperature, and 20 μL injection volume. Light had a wavelength of 279 nm. In Bogor, Indonesia, PT Mensana Aneka Satwa manufactured liquid antibiotics and combined them with broiler chicken tissue from 28-day-old antibiotic-free-fed chicks.

Results:
The tissue, liver, kidney, and muscle showed linearity values of R2=0.9999, R2=0.9992, and R2=0.9939 for norfloxacin and R2=0.9969, R2=0.9964, and R2=0.9938 for tylosin. Norfloxacin had liver, kidney, and muscle accuracy scores of 100.57%, 101.85%, and 105.41%, respectively, while tylosin had 103.89%, 95.83%, and 105.49%. Norfloxacin showed intraday precision values (%CV) of 0.77%, 4.18%, and 4% in the liver, kidney, and muscle, while tylosin had 0.74%, 1.84%, and 1%. Tylosin retained 3.325-3.358 minutes and norfloxacin 3.083-3.092. The LOD values for norfloxacin in the liver, kidney, and muscle were 1.57, 5.10, and 1.4 μg/mL, respectively. Tylosin levels in liver, kidney, and muscle were 1.72, 1.15, and 1.51 μg/mL, while norfloxacin had LOQ values of 5.24, 17.01, and 4.98 μg/mL.

Conclusion:
According to the European Medicines Agency Guideline, the mixed solution of norfloxacin and tylosin demonstrated favourable outcomes regarding sensitivity, specificity, linearity, accuracy, and precision.

Key words: Broiler tissue; HPLC; Norfloxacin-tylosin combination; Validation method.


 
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How to Cite this Article
Pubmed Style

Wibisono C, Wijayanti AD, Widiasih DA, Tjahajati I, Irianti IN, Pamudya KGS, Salsabila RAI, Sudimartini LM. Validation method for determining the concentration of norfloxacin-tylosin combination in broiler chicken tissue using high-performance liquid chromatography. Open Vet. J.. 2025; 15(9): 4153-4161. doi:10.5455/OVJ.2025.v15.i9.21

Web Style

Wibisono C, Wijayanti AD, Widiasih DA, Tjahajati I, Irianti IN, Pamudya KGS, Salsabila RAI, Sudimartini LM. Validation method for determining the concentration of norfloxacin-tylosin combination in broiler chicken tissue using high-performance liquid chromatography. https://www.openveterinaryjournal.com/?mno=260928 [Access: October 06, 2025]. doi:10.5455/OVJ.2025.v15.i9.21

AMA (American Medical Association) Style

Wibisono C, Wijayanti AD, Widiasih DA, Tjahajati I, Irianti IN, Pamudya KGS, Salsabila RAI, Sudimartini LM. Validation method for determining the concentration of norfloxacin-tylosin combination in broiler chicken tissue using high-performance liquid chromatography. Open Vet. J.. 2025; 15(9): 4153-4161. doi:10.5455/OVJ.2025.v15.i9.21


Vancouver/ICMJE Style

Wibisono C, Wijayanti AD, Widiasih DA, Tjahajati I, Irianti IN, Pamudya KGS, Salsabila RAI, Sudimartini LM. Validation method for determining the concentration of norfloxacin-tylosin combination in broiler chicken tissue using high-performance liquid chromatography. Open Vet. J.. (2025), [cited October 06, 2025]; 15(9): 4153-4161. doi:10.5455/OVJ.2025.v15.i9.21


Harvard Style

Wibisono, C., Wijayanti, . A. D., Widiasih, . D. A., Tjahajati, . I., Irianti, . I. N., Pamudya, . K. G. S., Salsabila, . R. A. I. & Sudimartini, . L. M. (2025) Validation method for determining the concentration of norfloxacin-tylosin combination in broiler chicken tissue using high-performance liquid chromatography. Open Vet. J., 15 (9), 4153-4161. doi:10.5455/OVJ.2025.v15.i9.21


Turabian Style

Wibisono, Cahyo, Agustina Dwi Wijayanti, Dyah Ayu Widiasih, Ida Tjahajati, Ika Nindya Irianti, Kanya Gabriella Sasri Pamudya, Ridha Avicena Ila Salsabila, and Luh Made Sudimartini. 2025. Validation method for determining the concentration of norfloxacin-tylosin combination in broiler chicken tissue using high-performance liquid chromatography. Open Veterinary Journal, 15 (9), 4153-4161. doi:10.5455/OVJ.2025.v15.i9.21


Chicago Style

Wibisono, Cahyo, Agustina Dwi Wijayanti, Dyah Ayu Widiasih, Ida Tjahajati, Ika Nindya Irianti, Kanya Gabriella Sasri Pamudya, Ridha Avicena Ila Salsabila, and Luh Made Sudimartini. "Validation method for determining the concentration of norfloxacin-tylosin combination in broiler chicken tissue using high-performance liquid chromatography." Open Veterinary Journal 15 (2025), 4153-4161. doi:10.5455/OVJ.2025.v15.i9.21


MLA (The Modern Language Association) Style

Wibisono, Cahyo, Agustina Dwi Wijayanti, Dyah Ayu Widiasih, Ida Tjahajati, Ika Nindya Irianti, Kanya Gabriella Sasri Pamudya, Ridha Avicena Ila Salsabila, and Luh Made Sudimartini. "Validation method for determining the concentration of norfloxacin-tylosin combination in broiler chicken tissue using high-performance liquid chromatography." Open Veterinary Journal 15.9 (2025), 4153-4161. Print. doi:10.5455/OVJ.2025.v15.i9.21


APA (American Psychological Association) Style

Wibisono, C., Wijayanti, . A. D., Widiasih, . D. A., Tjahajati, . I., Irianti, . I. N., Pamudya, . K. G. S., Salsabila, . R. A. I. & Sudimartini, . L. M. (2025) Validation method for determining the concentration of norfloxacin-tylosin combination in broiler chicken tissue using high-performance liquid chromatography. Open Veterinary Journal, 15 (9), 4153-4161. doi:10.5455/OVJ.2025.v15.i9.21


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