Xiao Kang Ping, Xiong Yuan, Liu Fang Zhu, Rustum Abu M
Global Quality Services - Analytical Sciences, Schering-Plough Corporation, Union, NJ 07083, USA.
J Chromatogr A. 2007 Sep 7;1163(1-2):145-56. doi: 10.1016/j.chroma.2007.06.027. Epub 2007 Jun 22.
In this paper, we describe a strategy that can be used to efficiently develop a high-performance liquid chromatography (HPLC) separation of challenging pharmaceutical molecules. This strategy involves use of advanced chromatographic technologies, such as a computer-assisted chromatographic method development tool (ChromSword) and an automated column switching system (LC Spiderling). This process significantly enhances the probability of achieving adequate separations and can be a large time saver for bench analytical scientists. In our study, the ChromSword was used for mobile phase screening and separation optimization, and the LC Spiderling was used to identify the most appropriate HPLC columns. For proof of concept, the analytes employed in this study are the structural epimers betamethylepoxide and alphamethylepoxide (also known as 16-beta methyl epoxide and 16-alpha methyl epoxide). Both of these compounds are used in the synthesis of various active pharmaceutical ingredients that are part of the steroid pharmaceutical products. While these molecules are relatively large in size and contain various polar functional groups and non-polar cyclic carbon chains, their structures differ only in the orientation of one methyl group. To our knowledge, there is no reported HPLC separation of these two molecules. A simple gradient method was quickly developed on a 5 cm YMC Hydrosphere C(18) column that separated betamethylepoxide and alphamethylepoxide in 10 min with a resolution factor of 3.0. This high resolution provided a true baseline separation even when the concentration ratio between these two epimers was 10,000:1. Although outside of the scope of this paper, stability-indicating assay and impurity profile methods for betamethylepoxide and for alphamethylepoxide have also been developed by our group based on a similar method development strategy.
在本文中，我们描述了一种可用于高效开发对具有挑战性的药物分子进行高效液相色谱（HPLC）分离的策略。该策略涉及使用先进的色谱技术，如计算机辅助色谱方法开发工具（ChromSword）和自动柱切换系统（LC Spiderling）。这一过程显著提高了实现充分分离的概率，对于实验室分析科学家而言可节省大量时间。在我们的研究中，ChromSword用于流动相筛选和分离优化，而LC Spiderling用于识别最合适的HPLC色谱柱。为了验证概念，本研究中使用的分析物是结构差向异构体倍他米松环氧物和α-甲基环氧物（也称为16-β-甲基环氧物和16-α-甲基环氧物）。这两种化合物都用于合成各种活性药物成分，这些成分是类固醇药物产品的一部分。虽然这些分子尺寸相对较大，含有各种极性官能团和非极性环状碳链，但它们的结构仅在一个甲基的取向上有所不同。据我们所知，尚无关于这两种分子的HPLC分离报道。在一根5 cm的YMC Hydrosphere C(18)色谱柱上快速开发了一种简单的梯度方法，该方法在10分钟内分离了倍他米松环氧物和α-甲基环氧物，分离度为3.0。即使这两种差向异构体的浓度比为10000:1时，这种高分离度也实现了真正的基线分离。尽管超出了本文范围，但我们小组基于类似的方法开发策略，也开发了倍他米松环氧物和α-甲基环氧物的稳定性指示测定法和杂质剖析方法。
