{"title":"HPLC-UV定量人血浆中CDK4/6抑制剂的新方法的建立与验证","authors":"Rakesh U. Shelke, Dinesh D. Rishipathak","doi":"10.2478/cipms-2023-0014","DOIUrl":null,"url":null,"abstract":"Abstract A sensitive and accurate high performance liquid chromatography method utilizing ultraviolet/visible light detection (HPLC-UV) for the quantification of Ribociclib in Spiked Human Plasma by HPLC-UV was developed and validated. Ribociclib (RCB) and the internal standard (IS), Trifluridine, were first extracted from plasma samples by a simple Protein Precipitation extraction using Acetonitrile. Plasma concentration of RCB and internal standard were then analyzed by applying reversed phase chromatography using Orochem orosil C18 (4.6 mm × 250 mm, 5 μ) and elution with a isocratic mobile phase consisting of 10 mM phosphate buffer – Acetonitrile (60:40, v/v) adjusted to pH 3.0 at a flow rate of 1.0 ml/min. Detection of RCB and the IS was subsequently done at a wavelength of 260 nm. The limit of quantification was 10 ng/ml. The calibration curve was linear (R2>0.998) over the concentration range of 10-1000 ng/ml. For human plasma, the accuracy and precision were within ±15% and ≤15%, respectively, for all concentrations, except for the lower limit of quantification, where they were within ≤20%.","PeriodicalId":11071,"journal":{"name":"Current Issues in Pharmacy and Medical Sciences","volume":"36 1","pages":"81 - 86"},"PeriodicalIF":0.4000,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"Development and validation of a new bioanalytical method for quantification of CDK4/6 inhibitor in Spiked Human Plasma by HPLC-UV\",\"authors\":\"Rakesh U. Shelke, Dinesh D. Rishipathak\",\"doi\":\"10.2478/cipms-2023-0014\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Abstract A sensitive and accurate high performance liquid chromatography method utilizing ultraviolet/visible light detection (HPLC-UV) for the quantification of Ribociclib in Spiked Human Plasma by HPLC-UV was developed and validated. Ribociclib (RCB) and the internal standard (IS), Trifluridine, were first extracted from plasma samples by a simple Protein Precipitation extraction using Acetonitrile. Plasma concentration of RCB and internal standard were then analyzed by applying reversed phase chromatography using Orochem orosil C18 (4.6 mm × 250 mm, 5 μ) and elution with a isocratic mobile phase consisting of 10 mM phosphate buffer – Acetonitrile (60:40, v/v) adjusted to pH 3.0 at a flow rate of 1.0 ml/min. Detection of RCB and the IS was subsequently done at a wavelength of 260 nm. The limit of quantification was 10 ng/ml. The calibration curve was linear (R2>0.998) over the concentration range of 10-1000 ng/ml. For human plasma, the accuracy and precision were within ±15% and ≤15%, respectively, for all concentrations, except for the lower limit of quantification, where they were within ≤20%.\",\"PeriodicalId\":11071,\"journal\":{\"name\":\"Current Issues in Pharmacy and Medical Sciences\",\"volume\":\"36 1\",\"pages\":\"81 - 86\"},\"PeriodicalIF\":0.4000,\"publicationDate\":\"2023-06-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current Issues in Pharmacy and Medical Sciences\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.2478/cipms-2023-0014\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"MEDICINE, GENERAL & INTERNAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Issues in Pharmacy and Medical Sciences","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.2478/cipms-2023-0014","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"MEDICINE, GENERAL & INTERNAL","Score":null,"Total":0}
引用次数: 1
摘要
摘要:建立了一种高效液相色谱紫外/可见光检测(HPLC-UV)定量加标人血浆中核糖西尼(Ribociclib)的方法。首先用乙腈进行简单的蛋白沉淀萃取,从血浆样品中提取核糖环尼(RCB)和内标物Trifluridine。采用Orochem orosil C18 (4.6 mm × 250 mm, 5 μ)反相色谱法分析血浆中RCB和内标物的浓度,以10 mm磷酸盐缓冲液-乙腈(60:40,v/v)为等温流动相,流速为1.0 ml/min,洗脱至pH 3.0。随后在260 nm波长下对RCB和IS进行检测。定量限为10 ng/ml。在10 ~ 1000 ng/ml的浓度范围内,校准曲线呈线性(R2>0.998)。对于人血浆,除定量下限在≤20%以内外,所有浓度的准确度和精密度分别在±15%和≤15%以内。
Development and validation of a new bioanalytical method for quantification of CDK4/6 inhibitor in Spiked Human Plasma by HPLC-UV
Abstract A sensitive and accurate high performance liquid chromatography method utilizing ultraviolet/visible light detection (HPLC-UV) for the quantification of Ribociclib in Spiked Human Plasma by HPLC-UV was developed and validated. Ribociclib (RCB) and the internal standard (IS), Trifluridine, were first extracted from plasma samples by a simple Protein Precipitation extraction using Acetonitrile. Plasma concentration of RCB and internal standard were then analyzed by applying reversed phase chromatography using Orochem orosil C18 (4.6 mm × 250 mm, 5 μ) and elution with a isocratic mobile phase consisting of 10 mM phosphate buffer – Acetonitrile (60:40, v/v) adjusted to pH 3.0 at a flow rate of 1.0 ml/min. Detection of RCB and the IS was subsequently done at a wavelength of 260 nm. The limit of quantification was 10 ng/ml. The calibration curve was linear (R2>0.998) over the concentration range of 10-1000 ng/ml. For human plasma, the accuracy and precision were within ±15% and ≤15%, respectively, for all concentrations, except for the lower limit of quantification, where they were within ≤20%.