A recent survey that was conducted by LCGC in late 2011 showed that since 2008, hydrophilic interaction liquid chromatography (HILIC) has more than doubled its application.
HILIC is finding more and more applications for polar compounds that are either unretained or weakly retained on reversed-phase
chromatographic columns. The technique also has been used for the separation of biomolecules. This article reviews the newer
stationary phases intended for HILIC and studies performed to categorize columns for optimized separations based on their
Hydrophilic interaction liquid chromatography (HILIC) has become an established chromatographic technique in widespread use
in most application areas. This great interest has lead to an accompanying increase in the number of new stationary phases
explored, some of which have become commercially available. For the user, this multiplication of phases is beneficial, but
it also presents some challenges. Where to start? What column to choose? How can one get optimal selectivity for a specific
application? Therefore, approaches to classify different HILIC stationary phases into categories have emerged. This article
will summarize the progress in this direction since a previous article was published on this topic in the 2008 supplement
Majors recently reported in an LCGC reader survey (2) that the use of HILIC mode had doubled since the year 2007. This means that roughly 25% of chromatographers
are using HILIC today. I had the pleasure to be responsible for the market introduction of the column brand ZIC-HILIC in January
2002, which was immediately well received by researchers at AstraZeneca. At that time, there were about 70 papers on HILIC
in the scientific literature published since the first paper by Alpert in 1990 (3). Back then few chromatographers had a complete
picture of the retention mechanism of HILIC or which column to use, but there was a multitude of unsolved application problems
that were delaying research in several branches of science. Most important, the orthogonal selectivity of HILIC to reversed-phase
chromatography was driving the users interest.
Later on, in April 2008 when the previous review in this series was written by McCalley (1) he had to consider in total 350
HILIC papers. Today, this number has risen to almost 1300 papers, with more than 300 new contributions only last year (see
Figure 1). Now, scientific literature has become a valuable source of separation applications using HILIC. The question of
whether separation is possible can often be answered based on the published work already done. More focus can be given to
how it is done and to other aspects of analysis such as sample preparation, throughput, and detection. Readers are referred
to special journal issues on HILIC (4–6), to a textbook with various chapters addressing successful applications (7), and
to reviews on different application areas such as pharmaceutical analysis (8), proteomics (9), metabonomic and metabolomic
analysis (10), glycomics (11), and bioanalysis (12).
It is clear that the improved understanding of HILIC and growing interest for selectivity, in general, also has led to better
sample handling by complementary pretreatment protocols and, in many methods, to an overall faster throughput.