Wollmer Erik, Klein Sandra
Department of Pharmacy, Institute of Biopharmaceutics and Pharmaceutical Technology, University of Greifswald, 17489 Greifswald, Germany.
J Pharm Pharm Sci. 2017;20(0):258-269. doi: 10.18433/J3K923.
A variety of fixed-dose combination products is used in the therapy of Parkinson Disease. However, to date a proper analytical method applicable for comparative screening of different antiparkinson products was not available. The objective of the present work was thus to develop and validate an analytical method for the simultaneous quantification of levodopa, carbidopa, benserazide and entacapone. The method should be applicable for quantifying samples from drug release experiments with marketed products and prototype formulations performed under compendial and biorelevant test conditions.
A fast and robust method applicable for separation and quantification of the four compounds was developed and validated according to International Conference on Harmonization guidelines. Method validation covered applicability to a wide concentration range of all compounds and peak separation in complex sample matrices such as biorelevant dissolution media.
The compounds were successfully separated by using a gradient elution method on an endcapped LiChrospher 100 RP-18 (250 x 4.6 mm, 5 µm) column coupled with a LiChrospher 100 RP-18 precolumn (4 x 4 mm, 5 µm) at a column temperature of 35.0 °C and a flow rate of 1.50 mL/min. The injection volume was 30 µL and the detection wavelengths were 280 and 210 nm, respectively. For all drug/media combinations the method was linear (r2 > 0.999) for a concentration range corresponding to 1.25 - 125 % label claim (i.e. 200 mg levodopa/entacapone and 50 mg carbidopa/benserazide) released. All other validation parameters were in the specified limits over the same concentration range.
The new method allows for robust and fast separation of levodopa, carbidopa, benserazide and entacapone without any interference caused by excipients or ingredients of compendial and biorelevant dissolution media and thus presents a valuable tool in both formulation development and in vitro drug release screening of numerous fixed-dose combinations of antiparkinson drugs. This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page.
多种固定剂量复方制剂用于帕金森病的治疗。然而，迄今为止，尚无适用于不同抗帕金森产品比较筛选的合适分析方法。因此，本研究的目的是开发并验证一种同时定量左旋多巴、卡比多巴、苄丝肼和恩他卡朋的分析方法。该方法应适用于在药典和生物相关试验条件下对市售产品和原型制剂进行药物释放实验所得样品的定量分析。
根据国际协调会议指南，开发并验证了一种适用于分离和定量这四种化合物的快速且稳健的方法。方法验证涵盖了对所有化合物宽浓度范围的适用性以及在复杂样品基质（如生物相关溶出介质）中的峰分离情况。
采用梯度洗脱法，在封端的LiChrospher 100 RP - 18（250×4.6 mm，5 µm）色谱柱与LiChrospher 100 RP - 18预柱（4×4 mm，5 µm）联用的条件下，于柱温35.0℃、流速1.50 mL/min成功分离了这些化合物。进样体积为30 µL，检测波长分别为280和210 nm。对于所有药物/介质组合，在对应于标示量1.25 - 125%（即200 mg左旋多巴/恩他卡朋和50 mg卡比多巴/苄丝肼）释放的浓度范围内，该方法呈线性（r2 > 0.999）。在相同浓度范围内，所有其他验证参数均在规定限度内。
该新方法能够稳健且快速地分离左旋多巴、卡比多巴、苄丝肼和恩他卡朋，不受辅料或药典及生物相关溶出介质成分的干扰，因此在众多抗帕金森药物固定剂量组合的制剂开发和体外药物释放筛选中均是一种有价值的工具。本文接受发表后审查。注册读者（见“致读者”）可通过点击本期目录页面上的摘要进行评论。
