Chaudhari Vishal Sharad, Borkar Roshan M, Murty Upadhyayula Suryanarayana, Banerjee Subham
Department of Pharmaceutics, National Institute of Pharmaceutical Education and Research (NIPER)-Guwahati, Changsari, 781101, Assam, India.
Department of Pharmaceutical Analysis, NIPER-Guwahati, Changsari, 781101, Assam, India.
J Pharm Biomed Anal. 2020 Jul 15;186:113325. doi: 10.1016/j.jpba.2020.113325. Epub 2020 Apr 29.
Quercetin and piperine are often used as an add-on therapy for various diseases, however both drug exhibits poor aqueous solubility and photosensitivity issue. Therefore, the aim of the present study is to improve the pharmaceutical challenges by incorporating both the drugs in nanostructured lipid carriers (NLCs) and to develop a sensitive, selective, accurate and precise reverse-phase high performance liquid chromatography (RP-HPLC) method for the simultaneous analysis of both drugs in NLCs. Effective chromatographic separation of quercetin and piperine was achieved on Hypersil gold C-18 column and mobile phase consisting of a mixture of acetonitrile and HPLC grade water (pH 2.6, adjusted with 2%v/v glacial acetic acid) in an isocratic elution mode. The flow rate of the mobile phase was 1 mL/min, column temperature at 35 ± 0.2 °C and the injection volume was 20 μL. The retention time for quercetin and piperine were found to be at 2.80 min and 10.36 min, respectively and detected at an isobestic wavelength of 346 nm using a photodiode array (PDA) detector. The method was found to be specific for the simultaneous analysis of quercetin and piperine in presence of NLCs matrix, accurate (>90%) and precise (%RSD < 2%). The validated RP-HPLC method effectively utilised to determine the percentage drug entrapment efficiency cum percentage drug loading of quercetin and piperine in NLCs enriched formulations along with the secondary estimation of in vitro cumulative percentage drug release study. The results were found to be reliable, hence the validated RP-HPLC method could be further used for the simultaneous detection and quantification of both these drugs in other lipid-based nano-formulations such as solid-lipid nanoparticles, polymer-lipid hybrid nanoparticles, lipid drug conjugates, etc. in in vitro and in vivo.
槲皮素和胡椒碱常被用作各种疾病的辅助治疗药物，然而这两种药物都存在水溶性差和光敏感性问题。因此，本研究的目的是通过将这两种药物载入纳米结构脂质载体（NLCs）来改善药学方面的挑战，并开发一种灵敏、选择性好、准确且精密的反相高效液相色谱（RP-HPLC）方法，用于同时分析NLCs中的这两种药物。在Hypersil gold C-18柱上实现了槲皮素和胡椒碱的有效色谱分离，流动相由乙腈和HPLC级水（pH 2.6，用2%v/v冰醋酸调节）的混合物组成，采用等度洗脱模式。流动相流速为1 mL/min，柱温为35±0.2℃，进样体积为20μL。槲皮素和胡椒碱的保留时间分别为2.80分钟和10.36分钟，使用光电二极管阵列（PDA）检测器在346 nm的等吸收波长处进行检测。该方法被发现对于在NLCs基质存在下同时分析槲皮素和胡椒碱具有特异性，准确（>90%）且精密（%RSD<2%）。经过验证的RP-HPLC方法有效地用于测定富含NLCs的制剂中槲皮素和胡椒碱的药物包封率及药物负载率百分比，以及体外累积药物释放研究的二次评估。结果被发现是可靠的，因此经过验证的RP-HPLC方法可进一步用于在体外和体内同时检测和定量其他基于脂质的纳米制剂（如固体脂质纳米粒、聚合物-脂质杂化纳米粒、脂质药物缀合物等）中的这两种药物。
