@article {510, title = {High-performance Thin-layer Chromatography Method Development, Validation, and Simultaneous Quantification of Four Compounds Identified in Standardized Extracts of Orthosiphon stamineus}, journal = {Pharmacognosy Research}, volume = {8}, year = {2016}, month = {August 2016}, pages = {238-243}, type = {Original Article}, chapter = {238}, abstract = {

Context: Orthosiphon stamineus is a medicinal herb widely grown in Southeast Asia and tropical countries. It has been used traditionally as a diuretic, abdominal pain, kidney and bladder inflammation, gout, and hypertension. Aims: This study aims to develop and validate the high-performance thin layer chromatography (HPTLC) method for quantification of rosmarinic acid (RA), 3{\textquoteright}-hydroxy-5,6,7,4{\textquoteright}-tetramethoxyflavone (TMF), sinensitin (SIN) and eupatorin (EUP) found in ethanol, 50\% ethanol and water extract of O. stamineus leaves. Materials and Methods: HPTLC method was conducted using an HPTLC system with a developed mobile phase system of toluene: ethyl acetate: formic acid (3:7:0.1) performed on precoated silica gel 60 F254 TLC plates. The method was validated based on linearity, accuracy, precision, limit of detection, limit of quantification (LOQ), and specificity, respectively. The detection of spots was observed at ultraviolet 254 nm and 366 nm. Results: The linearity of RA, TMF, SIN, and EUP were obtained between 10 and 100 ng/spot with high correlation coefficient value (R2) of more than 0.986. The limit of detection was found to be 122.47 {\textpm} 3.95 (RA), 43.38 {\textpm} 0.79 (SIN), 17.26 {\textpm} 1.16 (TMF), and 46.80 {\textpm} 1.33 ng/spot (EUP), respectively. Whereas the LOQ was found to be 376.44 {\textpm} 6.70 (RA), 131.45 {\textpm} 2.39 (SIN), 52.30 {\textpm} 2.01 (TMF), and 141.82 {\textpm} 1.58 ng/spot (EUP), respectively. Conclusion: The proposed method showed good linearity, precision, accuracy, and high sensitivity. Hence, it may be applied in a routine quantification of RA, SIN, TMF, and EUP found in ethanol, 50\% of ethanol and water extract of O. stamineus leaves.

}, keywords = {Eupatorin, High-performance thin-layer chromatography, Orthosiphon stamineus, Rosmarinic acid, Sinensitin, Validation}, doi = {10.4103/0974-8490.188872}, author = {Suzana Hashim and Hooi Kheng Beh and Mohamad Shahrul Ridzuan Hamil and Zhari Ismail and Amin Malik Shah Abdul Majid} } @article {393, title = {Analysis of L-citrulline and L-arginine in Ficus deltoidea leaf extracts by reverse phase high performance liquid chromatography}, journal = {Pharmacognosy Research}, volume = {7}, year = {2015}, month = {December 2014}, pages = {32-37}, type = {Original Article}, chapter = {32}, abstract = {

Background:\ Ficus deltoidea (FD) is one of the native plants widely distributed in several countries in Southeast Asia. Previous studies have shown that FD leaf possess antinociceptive, wound healing and antioxidant properties. These beneficial effects have been attributed to the presence of primary and secondary metabolites such as polyphenols, amino acids and flavonoids. Objective: The aim was to develop a reverse phase high-performance liquid chromatography method with ultraviolet detection that involves precolumn derivatisation with O-phthaladehyde for simultaneous analysis of two amino acids L-citrulline and L-arginine in FD leaf extracts. Materials and Methods: An isocratic elution program consisting of methanol: acetonitrile: Water at 45:45:10 v/v (solvent A) and 0.1 M phosphate buffer pH 7.5 (solvent B) at A: B v/v ratio of 80:20 on Zorbax Eclipse C18 SB-Aq column (250 {\texttimes} 4.6 mm, 5 {\textmu}m) were used. The flow rate was set at 1 ml/min and detection was carried out at 338 nm with 30 min separation time. Results: Good linearity for L-citrulline and L-arginine was obtained in the range 0.1-1000 {\textmu}g/ml at R2 >= 0.998. The limit of detection and limit of quantification values for both L-citrulline and L-arginine were 1 and 5 {\textmu}g/ml, respectively. The average of recoveries was in the range 94.94-101.95\%, with relative standard deviation (\%RSD) less than 3\%. Intra- and inter-day precision was in the range 96.36-102.43\% with RSD less than 2\%. Conclusion: All validation parameters of the developed method indicate the method is reliable and efficient for simultaneous determination of L-citrulline and L-arginine for routine analysis of FD.

}, keywords = {Ficus deltoidea, L-arginine, L-citrulline, Reverse phase high-performance liquid chromatography.}, doi = {10.4103/0974-8490.147196}, author = {Armaghan Shafaei and Abdalrahim F. A. Aisha and Mohammad Jamshed Ahmad Siddiqui and Zhari Ismail} } @article {384, title = {A novel reverse phase high-performance liquid chromatography method for standardization of Orthosiphon stamineus leaf extracts}, journal = {Pharmacognosy Research}, volume = {7}, year = {2015}, month = {December 2014}, pages = {23-31}, type = {Original Article}, chapter = {23}, abstract = {

Background: Orthosiphon stamineus Benth. (Lamiaceae) is a traditional medicinal plant which has been used in treating various ailments such as kidney diseases, bladder inflammation, arthritis and diabetes. The leaves contain high concentration of phenolic compounds, thus, rosmarinic acid (RA), 3{\textquoteright}-hydroxy-5, 6, 7, 4{\textquoteright}-tetramethoxyflavone (TMF), sinensetin (SIN) and eupatorin (EUP) were chosen as a marker compounds for standardization of various O. stamineus leaf extracts. Objectives: The aim was to develop and validate a new high-performance liquid chromatography (HPLC) method for quantification of 4 marker compounds (RA, TMF, SIN, EUP) in various O. stamineus leaf extracts. Materials and Methods: The method was developed and validated using RP-HPLC-diode-array detection at 320 nm for accuracy, precision and limits of detection and was applied for quantification of it markers in five different extracts prepared in solvents with increasing polarity, using a gradient mobile phase 0.1\% formic acid: Acetonitrile at a flow rate of 1 ml/min on reverse phase acclaim polar advantage II C18 column (3 {\textmu}m, 3 {\texttimes} 150 mm) with 18 min separation time. Results: The developed method provided satisfactory precision, and the accuracy of this method was in the range of 90.2\% to 105.5\%. All of 4 compounds showed good linearity at R2 \> 0.999. Conclusion: The developed method is a simple, cost effective with shorter run time (18 min) in comparison to previous methods (30 min) and utilization of environmental-friendly solvents system. Therefore, this method has the potential to replace currently used methods in the routine standardization work of O. stamineus extracts, raw materials and its commercial products.

}, keywords = {3{\textquoteright}-hydroxy-5, 4{\textquoteright}-tetramethoxyflavone, 6, 7, Eupatorin, Orthosiphon stamineus, Rosmarinic acid, Sinensetin.}, doi = {10.4103/0974-8490.147195}, author = {Noor Hafizoh Saidan and Abdalrahim F.A. Aisha and Mohd Shahrul Ridzuan Hamil and Amin Malik Shah Abdul Majid and Zhari Ismail} }