APL Research Centre-II (A Division of Aurobindo Pharma Ltd.), Hyderabad, India.
Department of Organic Chemistry, AU College of Science and Technology, Visakhapatnam, India.
J Sep Sci. 2022 Oct;45(19):3714-3724. doi: 10.1002/jssc.202200363. Epub 2022 Sep 11.
A new stability indicating reverse phase HPLC method has been developed and validated as per International Conference on Harmonization guidelines for the determination of sacubitril-valsartan premix stereoisomers, namely, (2R)-valsartan, (2S,4S)-sacubitril, (2R,4S)-sacubitril, and (2R,4R)-sacubitril. Primarily, stability indicating separation study was done on reverse phase LC conditions; it was described by peak homogeneity of sacubitril-valsartan and its stereoisomers. Cellulose tris(4-methylbenzoate) packing column Chiralcel OJ-RH(150 mm × 4.6 mm), 5 μm provided better resolution than those of amylose based stationary phase's. Resolution between two arbitrary adjacent analyte was found to be more than 2.0 with 0.1% trifluoroacetic acid in water as mobile phase-A and mobile phase-B consisting of acetonitrile, methanol, and trifluoroacetic acid (90:10:0.1, v/v/v). Gradient elution was performed at a flow rate of 1.0 ml/min, column temperature 20°C, injection volume 10 μl, UV detection at 254 nm and run time was 52 min. The detector response linearity of stereoisomers found to be linear (R ≥ 0.9998), limit of detection (0.290 μg/ml, 0.122 μg/ml, 0.123 μg/ml, and 0.124 μg/ml), and limit of quantification (0.878 μg/ml, 0.370 μg/ml, 0.373 μg/ml, and 0.375 μg/ml), respectively. Percentage recovery was found to be 98-105. Finally, the proposed method is user friendly and can be used in bulk drugs analysis.
已按照国际人用药品注册技术协调会（ICH）指导原则开发并验证了一种新的反相高效液相色谱（HPLC）法，用于测定沙库比曲缬沙坦预混立体异构体，即（2R）-缬沙坦、（2S，4S）-沙库比曲、（2R，4S）-沙库比曲和（2R，4R）-沙库比曲。首先，在反相 LC 条件下进行了具有稳定性指示的分离研究；它通过沙库比曲缬沙坦及其立体异构体的峰均一性来描述。纤维素三（4-甲基苯甲酸酯）填充柱 Chiralcel OJ-RH（150mm×4.6mm，5μm）提供的分辨率优于基于直链淀粉的固定相。发现任意两个相邻分析物之间的分辨率均大于 2.0，以水为流动相 A，流动相 B 由乙腈、甲醇和三氟乙酸（90:10:0.1，v/v/v）组成。以 1.0ml/min 的流速、20°C 的柱温、10μl 的进样量、254nm 的 UV 检测进行梯度洗脱，运行时间为 52min。立体异构体的检测器响应线性度被发现为线性（R≥0.9998），检测限（0.290μg/ml、0.122μg/ml、0.123μg/ml 和 0.124μg/ml）和定量限（0.878μg/ml、0.370μg/ml、0.373μg/ml 和 0.375μg/ml）。回收率为 98-105%。最后，该方法易于使用，可用于原料药分析。
