In the present study, pristine magnesium aluminium layered double hydroxide (Mg-Al LDH, sample A) nanoparticles were synthesized by simple co-precipitation technique followed by intercalation of methotrexate drug (MTX), their detailed physicochemical characterization and development of RP-HPLC method for estimation of MTX intercalated in Mg-Al LDH nanoparticles (sample B). The PXRD analysis showed the basal spacing of 003 plane corresponding to sample A and sample B to be 8.63 Å and 11.39 Å, respectively. Particle size distributions were measured to be 56-75 nm (D₅₀=66.15 nm) in case of sample A and 75-100 nm (D₅₀=87.41 nm) for sample B. Simple, accurate RP-HPLC method was developed for estimation of the MTX drug from sample B, on Agilent HPLC 1200 system with UV detector on L1 column. 0.02 M potassium dihydrogen phosphate solution (pH 3.0) and acetonitrile (85:15 v/v) was used as mobile phase in an isocratic elution mode at a flow rate of 1.0 mL/min at 35°C with a load of 10 μL at 307 nm. The retention time for methotrexate was at 4.44 min. The method was validated as per ICH Q2 (R1) guidelines. The method was found to be linear over a concentration range of 10-100 μg/mL.

在本研究中，通过简单的共沉淀技术合成了原始的镁铝层状双氢氧化物（Mg-Al LDH，样品 A）纳米粒子，然后插入甲氨蝶呤药物（MTX），详细的物理化学表征和 RP-HPLC 方法的发展用于估计插入到 Mg-Al LDH 纳米颗粒中的 MTX（样品 B）。PXRD 分析表明，对应于样品 A 和样品 B 的 003 平面的基础间距分别为 8.63 Å 和 11.39 Å。粒度分布经测定为56-75纳米（d ₅₀ = 66.15 nm）的样品A和75-100纳米的情况下（d ₅₀=87.41 nm) 样品 B。开发了简单、准确的 RP-HPLC 方法，用于在 L1 色谱柱上配备紫外检测器的 Agilent HPLC 1200 系统上估算样品 B 中的 MTX 药物。0.02 M 磷酸二氢钾溶液 (pH 3.0) 和乙腈 (85:15 v/v) 在等度洗脱模式下用作流动相，流速为 1.0 mL/min，35°C，上样量为 10 μL 307 纳米。甲氨蝶呤的保留时间为 4.44 分钟。该方法根据 ICH Q2 (R1) 指南进行了验证。发现该方法在 10-100 μg/mL 的浓度范围内呈线性。