Shafaei Armaghan, Aisha Abdalrahim F A, Siddiqui Mohammad Jamshed Ahmad, Ismail Zhari
Department of Pharmaceutical Chemistry, School of Pharmaceutical Sciences, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia.
Department of Kulliyyah of Pharmacy, International Islamic University Malaysia, Indera Mahkota Campus, Jalan Sultan Ahmad Shah, 25200 Kuantan, Pahang, Malaysia.
Pharmacognosy Res. 2015 Jan-Mar;7(1):32-7. doi: 10.4103/0974-8490.147196.
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.
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.
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 × 4.6 mm, 5 μm) were used. The flow rate was set at 1 ml/min and detection was carried out at 338 nm with 30 min separation time.
Good linearity for L-citrulline and L-arginine was obtained in the range 0.1-1000 μg/ml at R (2) ≥ 0.998. The limit of detection and limit of quantification values for both L-citrulline and L-arginine were 1 and 5 μg/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%.
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.
三角叶榕是广泛分布于东南亚多个国家的本土植物之一。先前的研究表明，三角叶榕叶具有抗伤害感受、伤口愈合和抗氧化特性。这些有益作用归因于多酚、氨基酸和黄酮类等一级和二级代谢产物的存在。
旨在开发一种带有紫外检测的反相高效液相色谱法，该方法涉及用邻苯二甲醛进行柱前衍生化，用于同时分析三角叶榕叶提取物中的两种氨基酸L-瓜氨酸和L-精氨酸。
采用等度洗脱程序，流动相为甲醇:乙腈:水（体积比45:45:10，溶剂A）和0.1 M pH 7.5的磷酸盐缓冲液（溶剂B），在Zorbax Eclipse C18 SB-Aq柱（250×4.6 mm，5 µm）上以A:B体积比80:20进行洗脱。流速设定为1 ml/min，检测波长为338 nm，分离时间为30 min。
L-瓜氨酸和L-精氨酸在0.1 - 1000 μg/ml范围内具有良好的线性关系，R²≥0.998。L-瓜氨酸和L-精氨酸的检测限和定量限分别为1和5 μg/ml。平均回收率在94.94 - 101.95%范围内，相对标准偏差（%RSD）小于3%。日内和日间精密度在96.36 - 102.43%范围内，RSD小于2%。
所开发方法的所有验证参数表明，该方法对于三角叶榕常规分析中同时测定L-瓜氨酸和L-精氨酸是可靠且高效的。
