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Article history:
Received 11 July 2014
Received in revised form 17 October 2014
Accepted 31 October 2014
Available online 8 November 2014
Keywords:
HPLC
Chromatographic simulation
ACD/Labs
Loratadine
Physico-chemical prediction
a b s t r a c t
Development of a robust HPLC method for pharmaceutical analysis can be very challenging and timeconsuming. In our laboratory, we have developed a new workow leveraging ACD/Labs software tools
to improve the performance of HPLC method development. First, we established ACD-based analytical
method databases that can be searched by chemical structure similarity. By taking advantage of the
existing knowledge of HPLC methods archived in the databases, one can nd a good starting point for
HPLC method development, or even reuse an existing method as is for a new project. Second, we used
the software to predict compound physicochemical properties before running actual experiments to
help select appropriate method conditions for targeted screening experiments. Finally, after selecting
stationary and mobile phases, we used modeling software to simulate chromatographic separations for
optimized temperature and gradient program. The optimized new method was then uploaded to internal
databases as knowledge available to assist future method development efforts. Routine implementation
of such standardized workows has the potential to reduce the number of experiments required for
method development and facilitate systematic and efcient development of faster, greener and more
robust methods leading to greater productivity. In this article, we used Loratadine method development
as an example to demonstrate efcient method development using this new workow.
2014 Elsevier B.V. All rights reserved.
1. Introduction
Analytical testing and control strategy plays a critical role
during the entire life cycle of the drug development process
in the pharmaceutical industry. HPLC is the major work horse
that has been used for all aspects of pharmaceutical analysis
including assay, dissolution analysis, impurity prole, forced degradation studies, process control, and drug metabolism studies
[14]. Given the time constraints and limited resources in an
R&D laboratory, it is imperative to develop robust HPLC methods
quickly to support the drug development process. Many different
approaches to improve the performance of HPLC method development have been reported [58]. Often, a column screening
system is used to nd a promising combination of mobile
phase/stationary phase which meets desired criteria, and the separation is subsequently optimized using a software tool such
as DryLab [712] or Chromsword [5,6,1316]. These software
tools allow the scientist to model chromatographic separations
based upon retention data from a limited number of scouting
Corresponding author. Tel.: +1 732 594 4515; fax: +1 732 594 3887.
E-mail address: jinjian.zheng@merck.com (J. Zheng).
http://dx.doi.org/10.1016/j.jpba.2014.10.032
0731-7085/ 2014 Elsevier B.V. All rights reserved.
50
L. Wang et al. / Journal of Pharmaceutical and Biomedical Analysis 104 (2015) 4954
L. Wang et al. / Journal of Pharmaceutical and Biomedical Analysis 104 (2015) 4954
51
Fig. 2. Structures of Loratadine (compound J) and its related compounds. Compound P is pseudoephedrine that is commonly used in combination with Loratadine. Compound
F is a pseudoephedrine related compound.
Fig. 3. Structures of compounds with similar structures to Loratadine found in the databases including protriptyline, nortriptyline, amitriptyline, doxepin, imipramine,
desipramine, trimipramine, and nordoxepin.
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L. Wang et al. / Journal of Pharmaceutical and Biomedical Analysis 104 (2015) 4954
Table 1
Summary of methods for similar structures from application database.
Compounds
Column
Mobile phase
Comments
Protriptyline, nortriptyline,
doxepin, imipramine,
amitriptyline
Protriptyline, nortriptyline,
doxepine, imipramine,
amitriptyline, trimipramine
Water/acetonitrile/TEA-phosphate, pH 6.9
Doxepine, trimipramine,
amitriptyline, imipramine,
nordoxepin, nortriptyline,
desipramine, protriptyline
Ascentis ES Cyano
Doxepine, protriptyline,
imipramine, nortriptyline,
amitriptyline, trimipramine
Hypersil Gold
Nordoxepin, protriptyline,
nortriptyline, imipramine,
amitriptyline
ZirChrom-PBD
Desmethyldoxepin, protriptyline,
esipramine, nortriptyline,
doxepine, imipramine,
amitriptyline, timipramine
Pursuit C18
Water/methanol/acetonitrile/potassium phosphate,
pH 7.0
Doxepine, imipramine,
nortriptyline, amitriptyline,
trimipramine
Nortriptyline, imipramine,
amitriptyline, clomipramine
XBridge C18
solution to 5.74 min for 0.05 mg/mL solution, and 5.81 min for
0.005 mg/mL solution (the lower the concentration, the longer
the retention time). Therefore, it is difcult to identify peaks
by RRT (relative retention time) as the retention time of active
pharmaceutical ingredient (API) varies with concentration. Clearly,
choice of appropriate buffer pH is critical to achieve a good peak
shape, highlighting the value of rst understanding basic analyte
physico-chemical properties prior to conducting generalized, broad
parameter screens to avoid collection of extensive data of limited
value.
L. Wang et al. / Journal of Pharmaceutical and Biomedical Analysis 104 (2015) 4954
53
We have presented an effective workow for method development employing a combination of hardware, software, focused
experimentation, and application of personal and organizational
knowledge. The new workow consists of three parts. First,
we established ACD-based HPLC method databases that can be
searched by chemical structure similarity to take advantage of the
existing knowledge of HPLC methods archived in the databases.
Second, we used software to predict compound physico-chemical
properties before running actual experiments to help select appropriate method conditions for targeted screening experiments. It is
worth noting that for Loratadine method development, screening
of stationary phase and mobile phases were not performed as all
impurities were well characterized, and a good starting condition
was found from the database. For complex samples where there
are lots of unknown impurities, targeted screening may be necessary to nd a promising starting condition. Finally, we use the
54
L. Wang et al. / Journal of Pharmaceutical and Biomedical Analysis 104 (2015) 4954
modeling software to simulate chromatographic separations, followed by targeted experimental verication in the laboratory, to
rapidly develop fast and robust HPLC methods. The optimized new
method was then added to the databases to assist future method
development. Such workow brings the benet of reduction in the
number of experiments required for method development. Faster,
greener and more robust methods are developed in a systematic and efcient manner, leading to productivity gains in overall
method development.
Acknowledgements
The authors would like to thank Thom Loughlin, Yong Liu,
Xiaodong Bu, and Patrick Chin for their support on Merck internal structure-searchable databases; and thank Mary Rogowski and
Karim Kassam from ACD/Labs for their support.
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