Comparative application of biological and ninhydrin- derivatized spectrophotometric assays in the evaluation and validation of amikacin sulfate injection
Abstract Instrumental techniques are preferred over bioassay methods for antibiotic quantification mainly due to speed and ability to quantify metabolites in biological samples; however, the potency and biological activity of these drugs cannot be assessed. Two methods - agar well diffusion (bio-ass...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Universidade de São Paulo
2023-01-01
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| Series: | Brazilian Journal of Pharmaceutical Sciences |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-82502022000100872&tlng=en |
| _version_ | 1797957107787497472 |
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| author | Edebi Nicholas Vaikosen Samuel Ogheneruona Origbo Diepreye Ere Prosper Odaderia |
| author_facet | Edebi Nicholas Vaikosen Samuel Ogheneruona Origbo Diepreye Ere Prosper Odaderia |
| author_sort | Edebi Nicholas Vaikosen |
| collection | DOAJ |
| description | Abstract Instrumental techniques are preferred over bioassay methods for antibiotic quantification mainly due to speed and ability to quantify metabolites in biological samples; however, the potency and biological activity of these drugs cannot be assessed. Two methods - agar well diffusion (bio-assay) and spectrophotometric methods were used to evaluate amikacin sulfate injection. Agar plates were inoculated with S. aureus inoculum; zones of inhibition from its susceptibility to amikacin were obtained, while spectrophotometric absorption at 650 nm of ninhydrin- derivatized amikacin in phosphate buffer (pH 8) was measured. Methods performance showed linearity from 1 - 16 μgmL-1 (bioassay, r = 0.9994) and 10-50 μgmL-1 (spectrophotometric, r = 0.9998). Molar absorptivity was 2.595 x 104 Lmol-1cm-1. Limits of detection and quantification were 1.07 and 3.24 μgmL-1 respectively for bioassay method, while corresponding values for spectrophotometric method were 0.98 and 2.97 μg mL-1. Relative standard deviations were ≤ 2.0% for both methods, with recoveries from 95.93 - 100.25%. Amikacin in brands ranged from 97.53 ± 2.68 to 100.84 ± 1.82%, student’s t-test was ≤ 2.78 (n = 4) with respect to label claim for both methods. Experimental paired t-test (t = 2.07; n = 4) and F-test (F = 3.94; n = 4) values indicated no significant difference between both methods, hence comparable and can jointly be used in quality control assessment of antibiotics. |
| first_indexed | 2024-04-10T23:59:06Z |
| format | Article |
| id | doaj.art-91c9b45abc464af4b1cd2647b5fcd926 |
| institution | Directory Open Access Journal |
| issn | 2175-9790 |
| language | English |
| last_indexed | 2024-04-10T23:59:06Z |
| publishDate | 2023-01-01 |
| publisher | Universidade de São Paulo |
| record_format | Article |
| series | Brazilian Journal of Pharmaceutical Sciences |
| spelling | doaj.art-91c9b45abc464af4b1cd2647b5fcd9262023-01-10T07:35:50ZengUniversidade de São PauloBrazilian Journal of Pharmaceutical Sciences2175-97902023-01-015810.1590/s2175-97902022e201185Comparative application of biological and ninhydrin- derivatized spectrophotometric assays in the evaluation and validation of amikacin sulfate injectionEdebi Nicholas Vaikosenhttps://orcid.org/0000-0002-2413-6127Samuel Ogheneruona OrigboDiepreye EreProsper OdaderiaAbstract Instrumental techniques are preferred over bioassay methods for antibiotic quantification mainly due to speed and ability to quantify metabolites in biological samples; however, the potency and biological activity of these drugs cannot be assessed. Two methods - agar well diffusion (bio-assay) and spectrophotometric methods were used to evaluate amikacin sulfate injection. Agar plates were inoculated with S. aureus inoculum; zones of inhibition from its susceptibility to amikacin were obtained, while spectrophotometric absorption at 650 nm of ninhydrin- derivatized amikacin in phosphate buffer (pH 8) was measured. Methods performance showed linearity from 1 - 16 μgmL-1 (bioassay, r = 0.9994) and 10-50 μgmL-1 (spectrophotometric, r = 0.9998). Molar absorptivity was 2.595 x 104 Lmol-1cm-1. Limits of detection and quantification were 1.07 and 3.24 μgmL-1 respectively for bioassay method, while corresponding values for spectrophotometric method were 0.98 and 2.97 μg mL-1. Relative standard deviations were ≤ 2.0% for both methods, with recoveries from 95.93 - 100.25%. Amikacin in brands ranged from 97.53 ± 2.68 to 100.84 ± 1.82%, student’s t-test was ≤ 2.78 (n = 4) with respect to label claim for both methods. Experimental paired t-test (t = 2.07; n = 4) and F-test (F = 3.94; n = 4) values indicated no significant difference between both methods, hence comparable and can jointly be used in quality control assessment of antibiotics.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-82502022000100872&tlng=enBiological activityPotencyDerivatizatonAminoglycosidesMolar absorptivityNinhydrin |
| spellingShingle | Edebi Nicholas Vaikosen Samuel Ogheneruona Origbo Diepreye Ere Prosper Odaderia Comparative application of biological and ninhydrin- derivatized spectrophotometric assays in the evaluation and validation of amikacin sulfate injection Brazilian Journal of Pharmaceutical Sciences Biological activity Potency Derivatizaton Aminoglycosides Molar absorptivity Ninhydrin |
| title | Comparative application of biological and ninhydrin- derivatized spectrophotometric assays in the evaluation and validation of amikacin sulfate injection |
| title_full | Comparative application of biological and ninhydrin- derivatized spectrophotometric assays in the evaluation and validation of amikacin sulfate injection |
| title_fullStr | Comparative application of biological and ninhydrin- derivatized spectrophotometric assays in the evaluation and validation of amikacin sulfate injection |
| title_full_unstemmed | Comparative application of biological and ninhydrin- derivatized spectrophotometric assays in the evaluation and validation of amikacin sulfate injection |
| title_short | Comparative application of biological and ninhydrin- derivatized spectrophotometric assays in the evaluation and validation of amikacin sulfate injection |
| title_sort | comparative application of biological and ninhydrin derivatized spectrophotometric assays in the evaluation and validation of amikacin sulfate injection |
| topic | Biological activity Potency Derivatizaton Aminoglycosides Molar absorptivity Ninhydrin |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-82502022000100872&tlng=en |
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