Salave Sagar, Jain Sonali, Shah Ravi, Benival Derajram
National Institute of Pharmaceutical Education and Research (NIPER), Department of Pharmaceutics, Ahmedabad 382355, India.
National Institute of Pharmaceutical Education and Research (NIPER), Department of Pharmaceutical Analysis, Ahmedabad 382355, India.
J AOAC Int. 2022 Dec 22;106(1):40-48. doi: 10.1093/jaoacint/qsac096.
Teriparatide is a recombinant fragment of human parathyroid hormone, a potent osteoanabolic agent used for osteoporosis.
The present study endeavored to develop a simple, rapid, and reliable reverse phase-high performance liquid chromatography (RP-HPLC) method for the determination of teriparatide in pegylated lipid nanovesicles for rapid formulation development/optimization.
A rapid RP-HPLC-based analytical method was developed for the quantification of teriparatide in pegylated lipid nanovesicles. The method was optimized on a Waters XBridge C18 (4.6 × 150 mm, 10 μm) column with a mobile phase consisting of 0.1% formic acid in water and acetonitrile both in a linear gradient program. In the method, a short run time of 9 min was achieved at a flow rate of 1.0 mL/min with an injection volume of 50 µL at a detection wavelength of 210 nm. The developed method was validated according to the ICH Q2 (R2) guideline. The method was applied for the quantification of teriparatide in prepared pegylated lipid nanovesicles. Teriparatide encapsulated pegylated lipid nanovesicles were prepared by the ethanol injection method. Further, these vesicles were characterized for % entrapment efficiency (%EE), particle size, zeta potential, and morphology by Cryo-SEM.
The teriparatide was eluting at 4.8 min in the run. Further, for the method validation, the linear relationship between concentration and response was established over the concentration range of 50-250 µg/mL with the R2 > 0.999. The method sensitivity was shown with LOD and LOQ with the value of 100 ng/mL and 500 ng/mL, respectively. The method was found to be accurate and precise with the recovery ranging in 100 ± 2% and RSD <2%, respectively. Minor deliberate changes proved the robustness of the developed method.
These results indicated that the developed and validated method is accurate, precise, rapid, reliable, and fit for the quantification of teriparatide in different formulations.
The RP-HPLC method was developed and validated for the quantification of teriparatide from novel pegylated lipid nanovesicles.
特立帕肽是一种重组人甲状旁腺激素片段，是一种用于治疗骨质疏松症的强效促骨合成药物。
本研究旨在开发一种简单、快速且可靠的反相高效液相色谱法（RP-HPLC），用于测定聚乙二醇化脂质纳米囊泡中的特立帕肽，以实现快速的制剂开发/优化。
开发了一种基于RP-HPLC的快速分析方法，用于定量聚乙二醇化脂质纳米囊泡中的特立帕肽。该方法在Waters XBridge C18（4.6×150 mm，10μm）色谱柱上进行优化，流动相由0.1%甲酸水溶液和乙腈组成，采用线性梯度洗脱程序。该方法在流速为1.0 mL/min、进样量为50μL、检测波长为210 nm的条件下，实现了9分钟的短运行时间。所开发的方法按照ICH Q2（R2）指南进行了验证。该方法应用于测定制备的聚乙二醇化脂质纳米囊泡中的特立帕肽。采用乙醇注入法制备了包封特立帕肽的聚乙二醇化脂质纳米囊泡。此外，通过冷冻扫描电子显微镜对这些囊泡的包封率（%EE）、粒径、zeta电位和形态进行了表征。
特立帕肽在运行过程中于4.8分钟洗脱。此外，为进行方法验证，在50-250μg/mL的浓度范围内建立了浓度与响应之间的线性关系，R2>0.999。该方法的灵敏度通过检测限（LOD）和定量限（LOQ）表示，分别为100 ng/mL和500 ng/mL。该方法的回收率在100±2%范围内，相对标准偏差（RSD）<2%，结果准确且精密。微小的故意改变证明了所开发方法的稳健性。
这些结果表明，所开发并验证的方法准确、精密、快速、可靠，适用于不同制剂中特立帕肽的定量分析。
开发并验证了RP-HPLC法用于定量新型聚乙二醇化脂质纳米囊泡中的特立帕肽。
