Publicação
DEVELOPMENT AND VALIDATION OF AN HPLC METHOD FOR THE DETERMINATION OF EIGHT CEPHALOSPORINS
| Resumo: | Cephalosporins are a class of antibiotics effective against a wide range of bacterial infections, making them widely used in the pharmaceutical industry. Hikma, specifically Hikma Sintra fa- cilities, is a company involved in the production of these antibiotics. The objective of this work was to develop and validate an analytical method capable of detecting all Cephalosporins pro- duced at Hikma. This method is intended to monitor and verify the effectiveness of cross- contamination prevention measures within the facilities. The current work developed an HPLC (High Pressure Liquid Chromatography) method, through several iterations, that successfully detects all eight Cephalosporins. The main variable while developing the method was the per- centage of organic, ACN (acetonitrile) or MeOH (methanol), in the different mobile phases attempted: 100 mM ammonium acetate buffer, 40 mM phosphate buffer and 10 mM sodium acetate buffer. The final method uses a XBridge BEH C18 5μm column at 32ºC, with a mobile phase consisting of 40 mM phosphate buffer, MeOH and ACN (82:13:5), at a flow rate of 1.2 mL/min and a UV detector set to 240 nm. The method demonstrated linearity with a correlation factor above 0.98 for all Cephalosporins, and precision, as indicated by a %RSD below 10% for all Cephalosporins, across six injections. The LOQ (Limit of Quantitation) and LOD (Limit of Detection) were determined to be 0.08 μg/mL and 0.02 μg/mL, respectively. The validated range of the method spans from 0.08 μg/mL to 0.5 μg/mL. Additionally, the swab recovery test confirmed the method's accuracy, with recovery percentages exceeding 75% for all Cephalo- sporins across the tested concentration range. |
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| Autores principais: | Santos, Gonçalo Leite |
| Assunto: | HPLC Contaminant Cephalosporins Validation Development |
| Ano: | 2024 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade Nova de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório Institucional da UNL |
| Resumo: | Cephalosporins are a class of antibiotics effective against a wide range of bacterial infections, making them widely used in the pharmaceutical industry. Hikma, specifically Hikma Sintra fa- cilities, is a company involved in the production of these antibiotics. The objective of this work was to develop and validate an analytical method capable of detecting all Cephalosporins pro- duced at Hikma. This method is intended to monitor and verify the effectiveness of cross- contamination prevention measures within the facilities. The current work developed an HPLC (High Pressure Liquid Chromatography) method, through several iterations, that successfully detects all eight Cephalosporins. The main variable while developing the method was the per- centage of organic, ACN (acetonitrile) or MeOH (methanol), in the different mobile phases attempted: 100 mM ammonium acetate buffer, 40 mM phosphate buffer and 10 mM sodium acetate buffer. The final method uses a XBridge BEH C18 5μm column at 32ºC, with a mobile phase consisting of 40 mM phosphate buffer, MeOH and ACN (82:13:5), at a flow rate of 1.2 mL/min and a UV detector set to 240 nm. The method demonstrated linearity with a correlation factor above 0.98 for all Cephalosporins, and precision, as indicated by a %RSD below 10% for all Cephalosporins, across six injections. The LOQ (Limit of Quantitation) and LOD (Limit of Detection) were determined to be 0.08 μg/mL and 0.02 μg/mL, respectively. The validated range of the method spans from 0.08 μg/mL to 0.5 μg/mL. Additionally, the swab recovery test confirmed the method's accuracy, with recovery percentages exceeding 75% for all Cephalo- sporins across the tested concentration range. |
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