| Research Article |
Open Access |
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| High Performance Liquid Chromatographic Method for the Determination of Clobetasol in Rat Plasma and its Application to Skin Penetration |
| Prashant B. Musmade1, Praful B. Deshpande1, Swapnil J. Dengle1, Kranti B. Talole1, Shriram M. Pathak1, Adiga MNS2, Krishnamurthy Bhat1, Nayanabhirama Udupa1 and K.Bairy2* |
| 1Dept. of Pharmaceutical Quality Assurance, Manipal College of Pharmaceutical Sciences, Manipal-576104 |
| 2Dept. of Pharmacology, Kasturba Medical College, Manipal-576104 |
| *Corresponding author: |
Dr. K. Laxminarayana Bairy,
Dept. of Pharmacology, Kasturba Medical College,
Manipal University, Manipal-576104, India,
Tel: +91 820 2922365,
Fax: +91 820 2922083,
E-mail: kl.bairy@manipal.edu. |
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| Received December 30, 2009; Accepted March 13, 2010; Published March 13, 2010 |
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| Citation: Musmade PB, Deshpande PB, Pathak SM, Adiga MNS, Bhat MK, et al. (2010) High Performance Liquid Chromatographic Method for the Determination of Clobetasol in Rat Plasma and its Application to Skin Penetration.J Bioanal Biomed 2: 001-007. doi: 10.4172/1948-593X.1000013 |
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| Copyright: © 2010 Musmade PB, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
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| Abstract |
| A simple and sensitive high performance liquid chromatography (HPLC) method was developed for quantification of clobetasol (CLB) in rat plasma. Aripiprazole was used as an internal standard (IS). The present method uses protein precipitation with acetonitrile, followed by the liquidliquid extraction with methyl tertiary butyl ether for extraction of the CLB from the matrix. Separation was carried out using Variance C18 (250.0 × 4.6 mm, 5 μm particle size) column and the effluent was monitored by an ultra violet (UV) detector at 240 nm. The mobile phase used was acetonitrile: phosphate buffer (pH 7.0; 25.0mM containing 0.2 %v/v triethylamine) (65: 35 % v/v) at a flow rate of 1.0 mL min-1. The present method was validated as per the United States Food and Drug Administration (USFDA) guideline. This method was linear over the range of 25.0- 1000.0 ng/ml with regression coefficient greater than 0.99. The mean recovery of CLB and IS were 70.8±1.9 and83.32±1.55 % respectively. The method was found to be precise, accurate and specific. It was applied for the estimation of CLB in plasma after topical application of various zinc salt in rats to study the penetration of CLB through rat skin. |
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