Rapid microbiological method for analysis of clindamycin-based product potency and its equivalence to UV and HPLC methods

IF 2.5 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of microbiological methods Pub Date : 2026-01-01 Epub Date: 2025-11-21 DOI:10.1016/j.mimet.2025.107338

Isadora Alves Lustosa, Ana Carolina Kogawa

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引用次数: 0

Abstract

Clindamycin is an antimicrobial sold in capsules, injectable solution, or gel for topical use. Despite its wide use, current official compendia and literature lack microbiological methods to assess its potency, relying mainly on physicochemical assays such as high-performance liquid chromatography (HPLC). These techniques, however, do not evaluate the biological activity of the drug. In this context, the present study proposes, for the first time, aimed to develop and validate a rapid microbiological method to determine the potency of clindamycin in capsule form and to compare its results with those obtained from UV and HPLC methods. The method employed Escherichia coli ATCC 8739, BHI broth as the growth medium, and a hydroalcoholic diluent (ethanol: water, 1:1 v/v) for solution preparation. Concentrations tested were 8, 16, and 32 μg mL⁻¹, with incubation at 35 ± 2 °C for 4 h under agitation (70 rpm), followed by absorbance measurement at 530 nm. The method exhibited linearity (r > 0.99), selectivity, precision (RSD 3.16 %, 3.57 %, 3.14 %), accuracy (average recovery of 99.47 %), and robustness under most tested conditions, as percentage of inoculum, brand of BHI broth, and volume of BHI broth used in the tube. Statistical comparison confirmed the equivalence of the proposed method with UV and HPLC (F_cal 0.68 < F_tab 5.14). This validated microbiological method offers a fast, reliable, and biologically relevant alternative for routine potency determination of clindamycin capsules, addressing a major analytical gap in the quality control of antimicrobial products.

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克林霉素产品效价的快速微生物学分析方法及其与紫外和高效液相色谱法的等效性。

克林霉素是一种抗菌剂，以胶囊、注射溶液或凝胶形式出售，用于局部使用。尽管其用途广泛，但目前的官方药典和文献缺乏微生物学方法来评估其效力，主要依赖于高效液相色谱（HPLC）等理化分析。然而，这些技术并不能评价药物的生物活性。在此背景下，本研究首次提出建立并验证一种快速测定克林霉素胶囊效价的微生物学方法，并将其结果与紫外法和高效液相色谱法进行比较。该方法以大肠杆菌ATCC 8739、BHI肉汤为生长培养基，以乙醇：水1:1 v/v的水醇稀释液配制溶液。测试浓度为8、16和32 μg mL-1，在35 ± 2 °C下孵育4 h，搅拌（70 rpm）， 530 nm处测定吸光度。在接种量、菌种、培养液体积等条件下，该方法均具有良好的线性（r > 0.99）、选择性、精密度（RSD 3.16 %、3.57 %、3.14 %）、准确度（平均回收率99.47 %）和稳健性。统计比较证实了该方法与UV和HPLC的等效性（Fcal 0.68 tab 5.14）。该方法为克林霉素胶囊的常规效价测定提供了一种快速、可靠、具有生物学相关性的替代方法，解决了抗菌产品质量控制的主要分析空白。

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来源期刊

Journal of microbiological methods 生物-生化研究方法

CiteScore

4.30

自引率

4.50%

发文量

151

审稿时长

29 days

期刊介绍： The Journal of Microbiological Methods publishes scholarly and original articles, notes and review articles. These articles must include novel and/or state-of-the-art methods, or significant improvements to existing methods. Novel and innovative applications of current methods that are validated and useful will also be published. JMM strives for scholarship, innovation and excellence. This demands scientific rigour, the best available methods and technologies, correctly replicated experiments/tests, the inclusion of proper controls, calibrations, and the correct statistical analysis. The presentation of the data must support the interpretation of the method/approach. All aspects of microbiology are covered, except virology. These include agricultural microbiology, applied and environmental microbiology, bioassays, bioinformatics, biotechnology, biochemical microbiology, clinical microbiology, diagnostics, food monitoring and quality control microbiology, microbial genetics and genomics, geomicrobiology, microbiome methods regardless of habitat, high through-put sequencing methods and analysis, microbial pathogenesis and host responses, metabolomics, metagenomics, metaproteomics, microbial ecology and diversity, microbial physiology, microbial ultra-structure, microscopic and imaging methods, molecular microbiology, mycology, novel mathematical microbiology and modelling, parasitology, plant-microbe interactions, protein markers/profiles, proteomics, pyrosequencing, public health microbiology, radioisotopes applied to microbiology, robotics applied to microbiological methods,rumen microbiology, microbiological methods for space missions and extreme environments, sampling methods and samplers, soil and sediment microbiology, transcriptomics, veterinary microbiology, sero-diagnostics and typing/identification.