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      A simple high performance liquid chromatography method for determination of rebamipide in rat urine

      research-article
      Author(s): , *
      Publication date (Electronic):
      Journal: MethodsX
      Publisher: Elsevier
      Keywords: Rebamipide, HPLC, Extraction, Fluorescence, Urine, Ofloxacin

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          Abstract

          Rebamipide is a mucoprotective agent commonly used to prevent nonsteriodal anti-inflammatory drug-induced gastrointenstinal side effects [1]. Human plasma and urine analysis of rebamipide utilizing high performance liquid chromatography (HPLC) have been reported [2]. Recently, we reported on the plasma levels of rebamipide in presense or absence of celecoxib or diclofenac in rats [3] using a modified HPLC method of detection developed by Jeoung et al. [4]. To tailor the method towards use in urinary rebamipide extraction and analysis, the following modifications were made:

          • •

            To compensate for high concentrations of rebamipide found in urine, a new rebamipide stock solution was prepared with a final concentration of 50,000 ng/mL.

          • •

            Rat urine calibration standards were obtained within the range of 50–1000 ng/mL and 1000–50,000 ng/mL.

          • •

            Plasma samples were replaced with urine samples.

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          Renal adverse effects of nonsteroidal anti-inflammatory drugs.

          Fakhreddin Jamali, Sam Harirforoosh (2009)
          NSAIDs depress prostaglandins synthesis through inhibition of COX-1 that is involved in maintaining cell integrity and COX-2 that, although presents particularly in the kidneys, is overexpressed in response to inflammation. Both the beneficial and side effects of NSAIDs are, therefore, through their inhibition of COX enzymes. Introduction of COX-2-selective inhibitors has improved the safety profile of the drugs with regard to their most common side effect which occurs at the gastrointestinal level but has not rendered them less cardio-nephrotoxic. Renal side effects of NSAIDs are rare, sometimes transient and often reversible upon drug withdrawal. The incident rate and the severity of the renal side effect, however, increase in patients with risk factors such as those with diabetes, heart failure, renal dysfunction and in the elderly. The side effects range from electrolyte retention and reduce glomerular filtration to nephritic syndrome and chronic renal failure. These effects are shared among NSAIDs with evidence of dose and exposure dependency. There is no known predictor for the nephrotoxicity. However, a relationship has been found between high plasma concentration and the renal adverse effect of NSAIDs. The usefulness of therapeutic drug monitoring in patients with risk factors needs to be explored.
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            Rebamipide: overview of its mechanisms of action and efficacy in mucosal protection and ulcer healing.

            A Tarnawski, T Yoshikawa, Tetsuo Arakawa … (1998)
            Rebamipide, a gastroprotective drug, is a compound selected from over 500 amino acid analogs of 2(1H)-quinolinone tested for gastroprotective action and for efficacy to heal experimental gastric ulcers. This drug stimulates prostaglandin generation in gastric mucosa and improves not only the speed but also the quality of ulcer healing. In addition, it protects the gastric mucosa against acute injury caused by various noxious and ulcerogenic factors. Based on these experimental results, rebamipide had been subsequently tested in several clinical trials and approved in Japan for therapeutic use in patients with gastric ulcers and patients with acute gastritis. The main purpose of developing this type of drug was to improve the quality of ulcer healing, especially in that antisecretory drugs lack this advantage. In a preliminary clinical study, rebamipide improved the quality of gastric ulcer healing and reduced future ulcer recurrence. A number of basic research studies have been performed to clarify the mechanisms of rebamipide's action. These studies demonstrated unique properties of rebamipide and convincingly showed that it increases gastric mucus glycoprotein components, stimulates migration and proliferation of wounded epithelial cell monolayers, increases expression of epidermal growth factor and its receptor in normal and ulcerated gastric mucosa, and scavenges active oxygen radicals. The drug also attenuates the activity of neutrophils and the production of inflammatory cytokines stimulated by NSAIDs and/or H. pylori. Therefore, rebamipide can contribute to the management of patients who are taking NSAIDs or are infected with H. pylori. The inhibition of immunoinflammatory responses by rebamipide in H. pylori-infected patients may prevent development of gastritis, peptic ulcer disease, its recurrence, and possibly gastric cancer. Moreover, rebamipide may enhance eradication of H. pylori-infection using standard eradication therapy. Further studies are needed to clarify these possible advantages of rebamipide.
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              Rebamipide inhibits indomethacin-induced small intestinal injury: possible involvement of intestinal microbiota modulation by upregulation of α-defensin 5.

              Kenji Watanabe, Toshio Watanabe, Masatsugu Shiba … (2013)
              Enterobacteria play important roles in the pathophysiology of small intestinal injuries induced by nonsteroidal anti-inflammatory drugs (NSAIDs). We investigated the effects of rebamipide, a gastrointestinal mucoprotective drug, on indomethacin-induced small intestinal injuries, intestinal microbiota, and expression levels of α-defensin 5, which is a Paneth cell-specific antimicrobial peptide and is important for the regulation of intestinal microbiota. Indomethacin (10mg/kg) was orally administered to mice after oral administration of rebamipide (100 or 300 mg/kg) or vehicle for 1 week, and the small intestinal injuries were assessed. After oral administration of rebamipide, the small intestinal contents were subjected to terminal restriction fragment length polymorphism (T-RFLP) analysis to assess the intestinal microbiota composition. Further, the expression levels of mRNA and protein for α-defensin 5 in the ileal tissue were determined by real-time reverse transcription-polymerase chain reaction and western blotting analysis, respectively. Rebamipide inhibited indomethacin-induced small intestinal injuries and T-RFLP analysis showed that rebamipide increased the percentage of Lactobacillales and decreased the percentage of Bacteroides and Clostridium than that in vehicle-treated controls. The mice that were treated with rebamipide showed an increase in α-defensin 5 mRNA expression and protein levels in the ileal tissue compared to vehicle-treated control mice. Indomethacin reduced expression of α-defensin 5 mRNA in ileal tissue, while rebamipide reversed expression of α-defensin 5 mRNA. In conclusion, our study results suggest that rebamipide inhibits indomethacin-induced small intestinal injuries, possibly by modulating microbiota in the small intestine by upregulation of α-defensin 5. Copyright © 2013 Elsevier B.V. All rights reserved.
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                Author and article information

                Contributors
                Sam Harirforoosh
                Journal
                Journal ID (nlm-ta): MethodsX
                Journal ID (iso-abbrev): MethodsX
                Title: MethodsX
                Publisher: Elsevier
                ISSN (Electronic): 2215-0161
                Publication date PMC-release: 30 June 2014
                Publication date Collection: 2014
                Publication date (Electronic): 30 June 2014
                Volume: 1
                Pages: 49-55
                Affiliations
                [0005]Department of Pharmaceutical Sciences, Gatton College of Pharmacy, East Tennessee State University, Johnson City, TN 37614, United States
                Author notes
                [* ]Corresponding author at: Department of Pharmaceutical Sciences, Gatton College of Pharmacy, East Tennessee State University, Box 70594, Johnson City, TN 37614-1708, United States. Tel.: +1 423 439 2074; fax: +1 423 439 6350. harirfor@ 123456etsu.edu
                Article
                Publisher ID: S2215-0161(14)20017-3
                DOI: 10.1016/j.mex.2014.06.002
                PMC ID: 4472849
                SO-VID: 12f21482-bff8-432f-869b-6361cd593cc8
                Copyright © © 2014 The Authors
                License:

                This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/).

                History
                Date received : 18 March 2014
                Date accepted : 5 June 2014
                Categories
                Subject: Article

                Keywords: rebamipide,hplc,extraction,fluorescence,urine,ofloxacin
                Data availability:
                Keywords: rebamipide, hplc, extraction, fluorescence, urine, ofloxacin

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