目的: 建立HPLC法检测盐酸普拉克索缓释片中有关物质。方法: 分别采用2种色谱条件进行有关物质测定检测。条件一：采用HC-5 C18（250 mm×4.6 mm,5 μm）色谱柱,以甲酸铵缓冲液（pH 5.0）-甲醇（99 ∶ 1）为流动相A,以甲酸铵缓冲液（pH 5.0）-甲醇（44 ∶ 56）为流动相B,梯度洗脱,流速1.0 mL · min-1,检测波长240、262、326 nm,柱温40 ℃,进样体积100 μL;条件二：采用Inertsil ODS-3（150 mm×3.0 mm,3 μm）色谱柱,以磷酸盐缓冲液（含0.5%辛烷磺酸钠,pH 3.0）-乙腈（90 ∶ 10）为流动相A,以磷酸盐缓冲液（含0.5%辛烷磺酸钠,pH 3.0）-乙腈（72 ∶ 28）为流动相B,梯度洗脱,流速0.7 mL · min-1,检测波长分别为240、262与326 nm,柱温40 ℃,进样体积100 μL。结果: 条件一能分离检测已知杂质12个,条件二能分离另外的已知杂质。盐酸普拉克索及其13个杂质的线性关系优异,各杂质的回收率（n=9）在95.0%~105.0%。最低检出限为2.0~13.0 ng · mL-1。强制性降解实验进一步验证了本文方法能够有效指示不同条件下的降解产物。采用条件一,检测出4批盐酸普拉克索缓释片中杂质1含量为0.07%~0.09%,杂质2含量为0%~0.01%,杂质11含量为0.04%,总杂质含量为0.11%~0.14%;采用条件二,检测出4批盐酸普拉克索缓释片中杂质2含量为0.01%~0.02%,杂质6的含量为0%~0.06%,杂质12的含量为0%~0.02%,未知总杂质的含量为0%~0.09%,总杂质含量为0.01%~0.16%。结论: 本研究建立的HPLC方法不仅优化了样品处理流程,而且显著提高了检测的准确性和可靠性,重现性良好,适用于盐酸普拉克索缓释片的质量控制。
Objective: To establish an HPLC method for the determination of related substances in pramipexole dihydrochloride sustained-release tablets. Methods: Two chromatographic methods were used to determine the related substances. Condition 1: HC-5 C18 (250 mm×4.6 mm, 5 μm) column was used, with ammonium formate buffer (pH 5.0)-methanol (99 ∶ 1) as mobile phase A and ammonium formate buffer (pH 5.0)-methanol (44 ∶ 56) as mobile phase B, the gradient elution was performedat a flow rate of 1.0 mL · min-1,the detection wavelengths were 240, 262 and 326 nm respectively, the column temperature was 40 ℃, and the injection volume was 100 μL. Condition 2: Inertsil ODS-3 (150 mm×3.0 mm, 3 μm) column was used, with phosphate buffer (containing 0.5% sodium octane sulfonate, pH 3.0)-acetonitrile (90 ∶ 10) as mobile phase A, and phosphate buffer (containing 0.5% sodium octane sulfonate, pH 3.0)-acetonitrile (72 ∶ 28) as mobile phase B, the gradient elution was performedat a flow rate of 0.7 mL · min-1, the detection wavelengths were 240, 262 and 326 nm respectively, the column temperature was 40 ℃, and the injection volume was 100 μL. Results: 12 known impurities were separated and detected under condition 1, while other known impurities were separated under condition 2. An excellent linear relationship was established between pramipexole dihydrochloride and its 14 impurities, and the recovery rate of each impurity (n=9) was determined to be 95.0%-105.0%. The minimum detection limit was found to be 2.0-13.0 ng · mL-1. The forced degradation experiment further confirmed that the degradation products under different conditions could be effectively indicated by this method. The results of testing four batches of pramipexole dihydrochloride sustained-release tablets showed that. Under condition 1, impurity 1 was present at 0.07%-0.09%, impurity 2 at 0%-0.01%, and impurity 11 at 0.04%, with total impurities at 0.11%-0.14%. Under condition 2, impurity 2 was found at 0.01%-0.02%, Impurity 6 at 0%-0.06%, impurity 12 at 0%-0.02%, and unknown total impurities at 0%-0.09%, with total impurities at 0.01%-0.16%. Conclusion: The HPLC method established in this study not only optimizes the sample preparation process, but also significantly improves the accuracy and reliability of detection, with good reproducibility, and making it suitable for the quality control of pramipexole dihydrochloride sustained-release tablets.
[1] PIERCEY MF.Pharmacology of pramipexole,a dopamine D3-preferring agonist useful in treating Parkinson’s disease[J].Clin Neuropharmacol,1998,21(3):141
[2] 陈生弟,陈海波.中国帕金森病治疗指南(第四版)[J].中华神经科杂志,2020,53(12):973
CHEN SD,CHEN HB.Chinese guidelines for the treatment of Parkinson’s disease(fourth edition)[J].Chin J Neurol,2020,53(12):973
[3] ARMSTRONG MJ,OKUN MS.Diagnosis and treatment of Parkinson disease:a review[J].JAMA,2020,323(6):548
[4] SCHAPIRA AH,BARONE P,HAUSER RA,et al.Extended-release pramipexole in advanced Parkinson disease:a randomized controlled trial[J].Neurology,2011,77(8):767
[5] MIZUNO Y,YAMAMOTO M,KUNO S,et al.Efficacy and safety of extended-versus immediate-release pramipexole in Japanese patients with advanced and L-dopa-undertreated Parkinson disease:a double-blind,randomized trial[J].Clin Neuropharmacol,2012,35(4):174
[6] 马义鹏,胡为民,梁蔚骏,等.普拉克索缓释片治疗原发性帕金森病非运动症状的临床观察[J].中国药物与临床,2017,17(8):1191
MA YP,HU WM,LIANG WJ,et al.Clinical observation of pramipexole extended release tablets intreating non-motor symptoms of primary Parkinson’s disease[J].Chin Rem Clin,2017,17(8):1191
[7] BENNETT JP Jr,PIERCEY MF.Pramipexole—a new dopamine agonist for the treatment of Parkinson’s disease[J].J Neurol Sci,1999,163(1):25
[8] 托马斯 · 弗里德尔,沃尔弗拉姆 · 艾森赖克.包含普拉克索或其药用盐的延长释放片剂:CN101005831B[P].2010-11-10
FRIEDEL T,EISENREICH W.Extended Release Tablets Containing Pramipexole or Its Medicinal Salts:CN101005831B[P].2010-11-10
[9] MITRA MS,DATTA K,HUTCHINSON R,et al.Harmonized 3Rs-based non-mutagenic impurity qualification study designs developed using the results of an IQ consortium survey[J].Regul Toxicol Pharmacol,2021,122:104895
[10] PAWAR SM,KHATAL LD,GABHE SY,et al.Establishment of inherent stability of pramipexole and development of validated stability indicating LC-UV and LC-MS method[J].J Pharm Anal,2013,3(2):9
[11] 李晶,石森林,杨继武.加校正因子的主成分自身对照法测定盐酸普拉克索的有关物质[J].中国现代应用药学,2015,32(11):1366
LI J,SHI SL,YANG JW.Determination of related substances of pramipexole hydrochloride by correction factor[J].Chin J Mod Appl Pharm,2015,32(11):1366
[12] 于峰,张曼红,刘建洋.HPLC法测定盐酸普拉克索的有关物质[J].山东化工,2020,49(11):67
YU F,ZHANG MH,LIU JY.Determination of related substances of pramipexole hydrochloride by HPLC[J].Shandong Chem Ind,2020,49(11):67
[13] 周益芬,林楠,宋光华,等.HPLC测定盐酸普拉克索缓释片中的有关物质[J].华西药学杂志,2015,30(2):245
ZHOU YF,LIN N,SONG GH,et al.Determination of related substances in pramipexole dihydrochloridesustained-release tablets by HPLC[J].West China J Pharm Sci,2015,30(2):245
[14] 王建科,周灵利,卢柳璇,等.HPLC测定盐酸普拉克索缓释片中的有关物质[J].海峡药学,2020,32(9):81
WANG JK,ZHOU LL,LU LX,et al.Determination of related substances in pramipexole dihydrochloride sustained-release tablets by HPLC[J].Strait Pharm J,2020,32(9):81
[15] 国家药典委员会编.国家药品标准工作手册:国家药品标准技术规范[M].第4版.北京:中国医药科技出版社,2013
Chinese Pharmacopoeia Commission.Work Book of National Drug Standards:Technical Specifications of National Drug Standards[M].4th Ed.Beijing:China Medical Science Press,2013
[16] BAKSHI M,SINGH S.Development of validated stability-indicating assay methods—critical review[J].J Pharm Biomed Anal,2002,28(6):1011
[17] 戈文兰,张根元.盐酸普拉克索缓释微丸胶囊的制备及有关物质检测[J].中国药房,2016,27(31):4433
GE WL,ZHANG GY.Preparation of pramipexole dihydrochloride sustained-release pellets capsules and related substances determination[J].China Pharm,2016,27(31):4433
[18] 李学林,王磊,刘辉,等.盐酸普拉克索缓释片有关物质的检测方法与流程:中国,CN202110249500.7[P].2021-05-28
LI XL,WANG L,LIU H,et al.Detection Method and Process of Related Substances of Pramipexole Hydrochloride Sustained-Release Tablets:China,CN202110249500.7[P].2021-05-28
