Development and Validation of an RP-HPLC Method for the Quantitative Analysis of Triclosan in Human Urine

Triclosan (TCS), a synthesized chlorinated phenolic compound, is commonly utilized in consumable products as an antimicrobial agent. TCS has sparked widespread awareness because of its toxicity and possible negative effect on public health in recent years. In this study, a highly sensitive, fast, and cost-effective isocratic reversed-phase high-performance liquid chromatography (RP-HPLC) method coupled with solid-phase extraction for analysis of triclosan in human urine samples was developed. The method utilized methanol and water in a ratio of 90 : 10 as the mobile phase on a Phenomenex Luna 3 µm C18(2) 100 Å, 150 × 4.60 mm stationary phase, with a runtime of 5 minutes. The method showed good resolution of triclosan in the presence of the sample matrix. Validation of the method was performed according to the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH). Linearity was tested over a range of 0.00625 µg/mL to 6.4 µg/mL, as accuracy recorded a recovery of 89.25%, 91.0%, and 92.75%. Limits of detection (LOD) and quantification (LOQ) were obtained to be 0.0173 µg/mL and 0.0525 µg/mL, respectively. The method proved to be robust over a temperature range of 26°C, 30°C, and 35°C and a flow rate of 0.5 ml, 1.0 ml, and 1.5 ml. The developed method was employed to detect and quantify triclosan in 153 urine samples, comprising 60 samples from Ibadan, Nigeria, and 93 samples from Kumasi, Ghana. Triclosan was detected in a total of 52 samples with an average content of 0.054588 µg/ml. This method can therefore be used for the routine analysis of triclosan in urine samples

HPLC-HRESI-MSn Characterization of Polyphenolic Compounds in the Stem Bark of Chlorophora regia A. Chev (Moraceae)

Isolation and identification of secondary metabolites from medicinal plants could be tedious and time consuming. Therefore, any technique that could be used to confirm the identity of medicinal plant constituents without isolating them will save time and resources. Chlorophora similar to many genera in the Moraceae family have been demonstrated to be rich sources of polyphenolic compounds with important biological activities. The current study was designed to employ HRESI-MSn analyses to qualitatively examine isolated polyphenolic compounds from the stem bark of Chlorophora regia. Based on the HRESI-MSn data obtained, the fragmentation patterns of the compounds under study will be proposed and could be used in their identification in a matrix. Five polyphenolic compounds were successfully isolated and purified using various chromatographic techniques including column chromatography, thin-layer chromatography (TLC) and preparative HPLC. The structures of the isolated compounds were elucidated by in-depth analyses of their 1D and 2D NMR and mass spectroscopic data. HRESI-MS/MS was further used to characterize the isolated compounds. Five polyphenolic compounds including three Diels-Alder type adducts: sanggenon C, kuwanol E and chalcomoracin; two stilbene derivatives: chlorophorin and isochlorophorin were isolated from the stem bark. The tandem MS fragmentation patterns of the compounds in positive mode, were successfully proposed. The fragments obtained and proposed fragmentation patterns of the isolated compounds could be employed qualitatively in the identification of the studied polyphenolic compounds in a matrix

Quality Control and Anti-Inflammatory Activity of the Stem Bark of Chlorophora regia A. Chev. (Moraceae): Quality Control of Herbal Products

This study sought to develop a validated reverse-phase high-performance liquid chromatography method for the quality control of the stem bark ingredients and its finished products and investigate the synergistic anti-inflammatory activity of the phytochemical constituents of C. regia stem bark. Fractionation and isolation of biomarkers were carried out by column chromatography on silica gel and monitored by thin-layer chromatography. The isolated biomarkers were characterized based on their melting points and extensive analysis of their spectroscopic data (IR, 1D and 2D NMR). The chromatographic separation was investigated and developed for the analysis of the biomarkers using µBondapakTM C18 (3.9×300 mm, 5 µm) as stationary phase. The mobile phase composition of 0.1 % trifluoroacetic acid as solvent A, and methanol as solvent B with gradient elution was finally selected. The carrageenan-induced edema in a 7-day-old chick model was used to assess the anti-inflammatory activity of the stem bark extract and compared to diclofenac sodium as a reference drug. Two compounds were successfully isolated and identified, as Regiafuran A (1) and 3,5,7,4’-Tetrahydroxy-2’-methoxyflavonol (2). The compounds were employed as biomarkers in the RP-HPLC method development. The developed method was validated and was successfully used to quantify the amount of 1 and 2 in the stem bark to be 0.224% ± 0.056%w/w and 0.354% ± 0.041%w/w respectively. The crude extract showed considerable anti-inflammatory activity compared to the reference drug, diclofenac. The method demonstrated acceptable levels of accuracy, precision, specificity, and robustness hence can be successfully adopted for routine quality control and standardization of the stem bark of C. regia. The stem bark of the plant exhibited significant anti-inflammatory activity

Acetic acid‐induced colitis modulating potential of total crude alkaloidal extract of Picralima nitida seeds in rats

Abstract Purpose The total crude alkaloidal extract of Picralima nitida seeds (PNE) is known to possess anti‐inflammatory activity among other therapeutic benefits although its benefits in colitis has not been investigated. The current study therefore seeks to investigate the anti‐colitis potential of PNE using acetic acid‐induced colitis model in rats. Methods Sprague Dawley rats were treated with oral 500 mg/kg sulphasalazine or 30, 100, and 300 mg/kg of PNE daily for 8 days with induction of colitis on the fourth day with acetic acid. Rats were killed 24 h after the last treatment and whole blood was obtained from the jugular vein for hematological analysis and biochemical assays. Colons were extirpated for assessment of macroscopic and histological damage to the colon. Results Treatment with PNE protected against colonic injury induced with acetic acid by decreasing mucosal ulceration, epithelial erosion, inflammatory cell infiltration, and colonic edema. Thus, PNE preserved mucosal architecture and suppressed goblet cells depletion. Moreover, treatment with PNE was associated with improved hematological parameters and reductions in the expression of serum tumor necrosis factor‐alpha, interleukin‐1β, and p38 mitogen‐activated protein kinase. Also, PNE treatment exerted antioxidant effects by reducing nitric oxide production and increasing glutathione levels. In addition, PNE inhibited colonic lipid peroxidation by decreasing myeloperoxidase activity and malondialdehyde production. Conclusion It can be concluded that PNE attenuates intestinal oxidative and inflammatory damages following intrarectal acetic acid challenge. Thus, demonstrates potential for use in chronic intestinal inflammatory diseases such as ulcerative colitis

Development and Validation of an RP-HPLC Method for the Quantitative Analysis of Triclosan in Human Urine

Funder: National Institute for Health Research; doi: http://dx.doi.org/10.13039/501100000272Triclosan (TCS), a synthesized chlorinated phenolic compound, is commonly utilized in consumable products as an antimicrobial agent. TCS has sparked widespread awareness because of its toxicity and possible negative effect on public health in recent years. In this study, a highly sensitive, fast, and cost-effective isocratic reversed-phase high-performance liquid chromatography (RP-HPLC) method coupled with solid-phase extraction for analysis of triclosan in human urine samples was developed. The method utilized methanol and water in a ratio of 90 : 10 as the mobile phase on a Phenomenex Luna 3 µm C18(2) 100 Å, 150 × 4.60 mm stationary phase, with a runtime of 5 minutes. The method showed good resolution of triclosan in the presence of the sample matrix. Validation of the method was performed according to the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH). Linearity was tested over a range of 0.00625 µg/mL to 6.4 µg/mL, as accuracy recorded a recovery of 89.25%, 91.0%, and 92.75%. Limits of detection (LOD) and quantification (LOQ) were obtained to be 0.0173 µg/mL and 0.0525 µg/mL, respectively. The method proved to be robust over a temperature range of 26°C, 30°C, and 35°C and a flow rate of 0.5 ml, 1.0 ml, and 1.5 ml. The developed method was employed to detect and quantify triclosan in 153 urine samples, comprising 60 samples from Ibadan, Nigeria, and 93 samples from Kumasi, Ghana. Triclosan was detected in a total of 52 samples with an average content of 0.054588 µg/ml. This method can therefore be used for the routine analysis of triclosan in urine samples