目的: 建立HPLC双波长法同时测定复方羟丙茶碱去氯羟嗪胶囊中二羟丙茶碱、盐酸去氯羟嗪、盐酸溴己新、消旋山莨菪碱、盐酸克伦特罗5个活性成分的含量。方法: 采用Zorbax Eclipse XDB-C18色谱柱(250 mm×4.6 mm, 5 μm),以0.01 mol·L-1磷酸二氢钾溶液(加离液盐:0.05 mol·L-1的高氯酸钠作为添加剂,磷酸调pH至2.5)为流动相A,乙腈为流动相B,梯度洗脱,流速1.0 mL·min-1,采用240 nm(二羟丙茶碱、盐酸去氯羟嗪、盐酸溴己新)和210 nm(消旋山莨菪碱、盐酸克伦特罗)的双波长作为检测波长,进样量20 μL,柱温35 ℃。结果: 二羟丙茶碱、盐酸去氯羟嗪、盐酸溴己新、消旋山莨菪碱、盐酸克伦特罗5个成分在2种波长下均达到完全分离;拖尾因子均<1.5;各成分质量浓度在相应范围内与峰面积呈良好线性关系;重复性RSD(n=6)分别为0.25%、0.50%、0.18%、0.77%、1.8%;各成分低、中、高3个加标水平下平均回收率(n=3)分别为100.7%~101.0%、100.0%~101.5%、100.4%~101.5%、99.8%~102.4%、97.6%~101.9%。3批样品中5个成分的含量分别为标示量的100.6%~101.4%、100.0%~100.9%、98.7%~98.9%、97.9%~98.2%、91.5%~92.4%。结论: 建立的方法可同时测定复方羟丙茶碱去氯羟嗪胶囊中全部5个活性成分,且方法简便准确,完善了其质量控制与评价体系;由于这5个成分也是其他止咳平喘类复方制剂中的常见活性成分,因此该方法也为其他复方制剂同时测定其中多个成分提供了借鉴。
Objective: To establish a double-wavelength HPLC method for simultaneous determination of all 5 active ingredients in diprophylline, bromhexine hydrochloride, decloxizine hydrochloride, raceanisodamine and clenbuterol hydrochloride capsules. Methods: The method was performed on a Zorbax Eclipse XDB-C18 column(250 mm×4.6 mm, 5 μm). The mobile phases consisted of A (0.01 mol·L-1 potassium dihydrogen phosphate solution with the chaotropic salt: 0.05 mol·L-1 sodium perchlorate as additive, adjusted to pH 2.5 with phosphoric acid) and B (acetonitrile) in gradient mode at a flow rate of 1.0 mL·min-1. The detective wavelength was set at 240 nm for diprophylline, bromhexine hydrochloride and decloxizine hydrochloride while 210 nm for raceanisodamine and clenbuterol hydrochloride. The injection volume was 20 μL and the column temperature was 35 ℃. Results: The five components of diprophylline, bromhexine hydrochloride, decloxizine hydrochloride, raceanisodamine and clenbuterol hydrochloride were separated well at the both wavelength with the tailing factor below 1.5 and indicated good linearity between peak areas and concentration. RSD of the repeatability tests (n=6) were 0.25%, 0.50%, 0.18%, 0.77% and 1.8% while the average recoveries (n=3) at the low, medium and high spiked levels were 100.7%-101.0%, 100.0%-101.5%, 100.4%-101.5%, 99.8%-102.4% and 97.6%-101.9% respectively. The contents of the five components in three batches of samples were 100.6%-101.4%, 100.0%-100.9%, 98.7%-98.9%, 97.9%-98.2% and 91.5%-92.4% of the labeled amount respectively. Conclusion: This method proved simple and accurate for simultaneous determination of all five active ingredients, which can be used for quality control and evaluation of this preparation. The method also provides an example to simultaneous determination for the relative active ingredients in other preparations since the five components were common in cough medicine.
[1] 国家药品标准: 化学药品地方标准上升国家标准.第5册[S]. 2002: 167
National Drug Standards: National Standards for Chemicals Promoted from Local Standards. Vol 5[S]. 2002: 167
[2] 茆玉国, 杨波. RP-HPLC法测定舒喘平胶囊中3个主要成分的含量[J].药学与临床研究, 2007, 15(2): 113
MAO YG, YANG B. Determination of diprophylline, bromhexine hydrochloride and decloxizine hydrochloride in Shuchuanping capsules by HPLC[J].Pharm Clin Res, 2007, 15(2): 113
[3] 周震宇. 复方羟丙茶碱去氯羟嗪胶囊中盐酸克伦特罗的含量测定[J].中国药品标准, 2017, 18(1): 25
ZHOU ZY. Determination of clenbuterol in diprophylline bromhexine hydrochloride decloxizine hydrochloride raceanisodamine and clenbuterol hydrochloride capsules[J].Drug Stand China, 2017, 18(1): 25
[4] 尹清, 李文浩, 徐宏祥, 等. HPLC法同时测定茶新那敏薄膜包衣片中三组分的含量[J].药物分析杂志, 2012, 32(10): 1861
YIN Q, LI WH, XU HX, et al. HPLC simultaneous determination of three kinds of ingredients in aminophylline bromhexine and chlorphenamine membrane coat tablets[J].Chin J Pharm Anal, 2012, 32(10): 1861
[5] 孙树周, 何燕莉, 陈泳恩, 等. 液相色谱-串联质谱法测定凉茶中非法添加的26种止咳平喘类化学药物[J].今日药学, 2021, 31(3): 195
SUN SZ, HE YL, CHEN YE, et al. Determination of 26 chemical drugs for the treatment of cough and asthma added illegally in herbal tea by high performance liquid chromatography-tandem mass spectrometry[J].Pharm Today, 2021, 31(3): 195
[6] 夏方亮, 王玉林, 王建强, 等. 止咳平喘类制剂中可能违禁添加的14种药物的UPLC-DAD法检测[J].中国医药工业杂志, 2018, 49(5): 651
XIA FL, WANG YL, WANG JQ, et al. Determination of fourteen illegally added drugs in anti-cough and antiasthmatic preparations by UPLC-DAD[J].Chin J Pharm, 2018, 49(5): 651
[7] 李奕婧, 李晓琳, 韩家忠, 等. HPLC-DAD-MS/MS检测止咳平喘类中药制剂中非法添加的12种化学药品[J].药学学报, 2017, 52(6): 959
LI YJ, LI XL, HAN JZ, et al. Screening for 12 mixed illegally cough-relieving chemical composition in anti-cough and antiasthmatic traditional Chinese medicines by HPLC-DAD-MS/MS[J].Acta Pharm Sin, 2017, 52(6): 959
[8] 王瑞芬, 郭海丽. HPLC同时测定喘舒片中的9种成分[J].华西药学杂志, 2021, 36(1): 69
WANG RF, GUO HL. Simultaneous determination of nine constituents in Chuanshu tablets by HPLC[J].West China J Pharm Sci, 2021, 36(1): 69
[9] 中华医学会呼吸病学分会哮喘学组. 支气管哮喘防治指南(2020年版)[J].中华结核和呼吸杂志, 2020, 43(12): 1023
Asthma group of Chinese Throacic Society. Guidelines for bronchial asthma prevent and management (2020 edition)[J].Chin J Tubere Respir Dis, 2020, 43(12): 1023
[10] 李文喆, 吴茜, 董雪峰, 等. 呼吸科门诊清热止咳平喘类中成药用药分析[J].中国医药导报, 2021, 18(20): 124
LI WZ, WU Q, DONG XF, et al. Analysis of Chinese patent medicine for clearing heat, relieving cough and relieving asthma in respiratory outpatient[J].China Med Her, 2021, 18(20): 124
[11] SHULYAK N, PIPONSKI M, KOVALENKO S, et al. Chaotropic salts impact in HPLC approaches for simultaneous analysis of hydrophilic and lipophilic drugs[J].J Sep Sci, 2021,44(15): 2908
[12] RMANDI M, MALENOVI A. Chaotropic chromatography method development for the determination of aripiprazole and its impurities following analytical quality by design principles[J].J Sep Sci, 2020, 43(16): 3242
[13] ZHONG JJ, LIAO NB, HE MF, et al. Development of an analytical method for urocanic acid isomers in fish based on reactive extraction cleanup and chaotropic chromatography techniques[J].J Chromatogr A, 2018, 1548: 44
[14] AHMAD I, BENNETT R, MAKEY D, et al. In silico method development for the reversed-phase liquid chromatography separation of proteins using chaotropic mobile phase modifiers[J].J Chromatogr B, 2021, 1173: 122587
[15] 中华人民共和国药典2020年版. 二部[S]. 2020: 1325
ChP 2020. Vol Ⅱ[S]. 2020: 1325
