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Skin sample preparation by collagenase digestion for diclofenac quantification using LC-MS/MS after topical
Ramakrishna Nirogi1, Naga Surya Prakash Padala1, Rajesh Kumar Boggavarapu1
1Biopharmaceuticals Research, Suven Life Sciences Ltd, Serene Chambers, Road-5, Avenue-7, Banjara Hills, Hyderabad 500034, India.
Bioanalysis
|May 27, 2016
Summary
A new method using collagenase and bead beating efficiently prepares skin samples for drug analysis. This technique ensures uniform homogenates for reproducible pharmacokinetic studies of topical applications.
Area of Science:
- Pharmacology
- Bioanalytical Chemistry
- Dermatology
Background:
- Skin is a key site for topical drug pharmacokinetic evaluation.
- Effective sample preparation is crucial for accurate bioanalysis of drugs in skin.
- Current methods for preclinical dermatopharmacokinetics lack efficiency.
Purpose of the Study:
- To develop an efficient sample preparation method for skin bioanalysis.
- To enable accurate quantification of drugs in skin using LC-MS/MS.
Main Methods:
- Enzymatic treatment of skin samples with collagenase.
- Mechanical homogenization using a bead beater.
- Evaluation of the combined technique for sample preparation.
Main Results:
- The collagenase treatment followed by bead beating homogenization yielded uniform skin homogenates.
- This method demonstrated reproducibility for bioanalytical results.
- The optimized procedure is suitable for preclinical dermatopharmacokinetic studies.
Conclusions:
- A novel approach combining enzymatic and mechanical methods improves skin sample preparation.
- This technique facilitates efficient bioanalysis and drug quantification in skin.
- The developed method addresses the need for efficient preclinical dermatopharmacokinetic sample processing.
