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Journal of Pharmaceutical Research International, ISSN: 2456-9119, ISSN: 2231-2919 (past),Vol.: 21, Issue.: 4


Determination of Sodium (2-(2, 6-dichloroanilino) phenyl) Acetic Acid in Human Plasma by Rapid and Sensitive HPLC Method and UV-Spectrophotometry: Its Comparative Evaluation


Saeed-ul-Hassan1, Javaria Chishti2, Kashif Barkat1*, Hammad Yousaf1 and Asif Mahmood1

1Faculty of Pharmacy, University of Lahore, Lahore -54000, Punjab, Pakistan.

2University College of Pharmacy, Punjab University, Lahore, Pakistan.

Article Information
(1) Rahul S. Khupse, Pharmaceutical Sciences, University of Findlay, USA.
(2) Syed A. A. Rizvi, Department of Pharmaceutical Sciences, College of Pharmacy, Nova Southeastern University, USA.
(1) Mohammed Gamal Mahmoud, Aljouf University, Saudia Arabia.
(2) Haroon Rahim, Sarhad University of Science and Information Technology, Pakistan.
(3) Ana Carolina Kogawa, Universidade Estadual Paulista, Brazil.
Complete Peer review History: http://www.sciencedomain.org/review-history/23570


Aim: The objectives of current study were to develop and validate a simple, rapid, and sensitive HPLC method for determination of sodium (2-(2, 6- dichloroanilino) phenyl) acetic acid (SDPAA) in human plasma according to US-FDA guidelines and determination of SDPAA by UV- Spectrophotometry.

Methodology: In case of HPLC method, the mobile phase composed a mixture of acetonitrile (ACN) and phosphate buffer (pH 6.8) in a ratio of 40:60 ml (pH 3.5). A Shimadzu HPLC machine (HPLC 10 ATVP) with a column Chromolith-R high resolution RP–18 end caped and a column length of 100mm to 4.6 mm, with a UV detector was used. The peak was observed at wavelength of 281 nm. The sample was injected at a flow rate of 1.5 ml per min. The solvent run time was 8 minutes and the average retention time of the six sample observed was 5.66 minutes. In case of UV-spectrophotometric method, Shimadzu UV-1800 spectrophotometer was used. Mixture of phosphate buffer of pH 6.8:ACN was selected as a solvent for determination of SDPAA at 281 nm. Beer law was obeyed in the range of 2-22 µg/ml. The results of both UV-spectrophotometric and HPLC methods in determination SDPAA, were compared.

Results: SDPAA was detected at 281nm and eluted at 5.669 min (without plasma) while the samples extracted from the plasma eluted at 5.667 min. The average % RSD was less than 2%. Accuracy was confirmed with the recovery studies and by three test assays. Accuracy was tested at three %age level that is within 95–99%. The linear range of 0.5-20 µg/ml (for HPLC) showed the regression coefficient (R2) 0.999 and linear range of 2-22 µg/ml (for UV method) showed the regression coefficient (R2) 0.998 which were in acceptable range.

Conclusion: The developed HPLC method was simple, rapid, reliable, specific and sensitive with ability to determine drug concentrations from human plasma.

Keywords :

SDPAA; HPLC method; human plasma and UV-spectrophotometry.

Full Article - PDF    Page 1-15

DOI : 10.9734/JPRI/2018/39301

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