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Published in final edited form as:

J Sep Sci. 2020 May ; 43(9-10): 1623–1627. doi:10.1002/jssc.201901276.

Portable Capillary Liquid Chromatography for Pharmaceutical

and Illicit Drug Analysis
Samuel W. Foster1, Xiaofeng Xie2, Michelle Pham1, Paul A. Peaden2, Leena M. Patil3, Luke
T. Tolley2, Paul B. Farnsworth3, H. Dennis Tolley4, Milton L. Lee2,3, James P. Grinias1,*
1Department of Chemistry & Biochemistry, Rowan University, Glassboro, NJ 08028
2Axcend Corporation, Provo, UT 84604
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3Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602

4Department of Statistics, Brigham Young University, Provo, UT 84602

Abstract
A newly developed portable capillary liquid chromatograph was investigated for the separation of
various pharmaceutical and illicit drug compounds. The system consists of two high-pressure
syringe pumps capable of delivering capillary-scale flow rates at pressures up to 10,000 psi.
Capillary LC columns packed with sub-2 μm particles are housed in cartridges that can be inserted
into the system and easily connected through high-pressure fluidic contact points by simply
applying a specific, pre-determined torque rather than using standard fittings and less precise
sealing protocols. Several over-the-counter analgesic drug separations are demonstrated, along
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with a simple on-line measurement of tablet dissolution. Twenty illicit drug compounds were also
separated across six targeted drug panels. The results described in this study demonstrate the
capability of this compact LC instrument to address several important drug-related applications
while simplifying system operation, and greatly reducing solvent usage and waste generation
essential for on-site analysis.

Keywords
capillary liquid chromatography; portable; pharmaceuticals; illicit drugs; dissolution monitoring

1. Introduction
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Liquid chromatography (LC) continues to be one of the most important techniques for the
chemical analysis of pharmaceutical, forensic, food and beverage, environmental,
biomedical, and clinical samples. Although commercial LC instrumentation has gradually
improved over the past five decades, except for the pronounced development of ultrahigh

*
Corresponding Author: James P. Grinias, Rowan University, 201 Mullica Hill Rd., Glassboro, NJ 08028, grinias@rowan.edu, Phone:
+1-856-256-5461, Fax: +1-856-256-4478.
Conflict of Interest Statement
XX, PAP, MLL, LTT, PBF and HDT are associated (either as employees or consultants) with the company (Axcend Corporation) that
is developing the technology described in this manuscript for commercialization.
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pressure liquid chromatography (UHPLC) since 2004 [1], the general approach to the
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technique has remained fundamentally the same over this time period. In contrast to gas
chromatography (GC), the use of capillary-scale columns has not dominated the LC field
during its maturation. Rather, capillary LC has been primarily utilized in ‘omics’
applications because of its enhanced sensitivity when interfaced to mass spectrometry (MS)
as compared to analytical-scale columns operated at much higher flow rates [2–4]. The key
advantages of capillary LC include (1) reduced flow rate, which makes the method greener
compared to analytical-scale LC, and (2) improved heat dissipation, which eliminates the
effects of viscous friction when coupled with lower flow rates [5,6]. However, concerns
about column robustness and reduced sensitivity using optical detection modes have limited
the use of capillary LC much beyond the aforementioned ‘omics’ studies [7,8].

In this Highlights article, a novel approach to capillary LC instrument design is introduced

to alleviate some of these concerns. To realize this new design, advances in small, high-
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pressure LC pumps [9–11] and on-column UV-absorbance detection (LC-UV) [12,13] have
been integrated into a compact instrument footprint that is field-portable [14]. Column
robustness issues and extra-column band broadening related to poor connections in capillary
systems are reduced through the use of a novel cartridge design that applies appropriate
torque for fluidic connections at pressures up to 10,000 psi. This is in contrast to other
portable LC systems that employ more traditional LC column integration and lower pressure
limits [15–18]. This first report demonstrating the use of the new, compact LC system
addresses several common applications in the pharmaceutical and forensic fields.

2. Discussion
2.1 Description of the Compact Capillary LC Platform
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An overall schematic of the instrument design of the portable capillary LC platform is

shown in Figure 1. The system is 20.1 cm (height) × 23.1 cm (width) × 32.0 cm (depth) and
weighs 7.82 kg (a size comparison to other portable LC systems can be found in the
Supporting Information). Two high-pressure syringe pumps capable of delivering flow rates
of 0.8–50 μL/min at pressures up to 10,000 psi are operated in tandem and connected to a
mixing valve so that a single mobile phase with combined solvents from two reservoirs is
delivered to the injection valve (either in isocratic or gradient operating modes). The injector
contains an internal sample loop with 40 nL volume (this value can be decreased to as low as
4 nL with a different internal loop or increased up to 700 nL with a slightly modified valve),
which follows a mobile phase mixing valve and connects to the cartridge through a short
section of small-diameter (i.e., 75 μm i.d.) tubing. The capillary LC column (fused silica
with a UV-transparent outer coating packed with sub-2 μm stationary phase particles) is
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mounted inside a cartridge that also contains an on-capillary UV-absorbance detector with a
light-emitting diode (LED) light source [13,19]; the inlet of the capillary forms a high-
pressure fluidic connection to the tubing from the injector when torque is applied to the
cartridge mechanism from a calibrated screw knob on the front of the instrument. In this
study, the cartridges contained 100 mm × 150 μm i.d. capillary columns packed with sub-2
μm C18 fully porous particles. Applications that are commonly performed with traditional

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LC instrumentation were translated to the capillary scale and performed using the integrated,
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compact platform.

2.2 Use of the Compact Capillary LC Platform for Pharmaceutical Separations

One key area that regularly employs LC separations coupled to UV-absorbance detection is
the analysis of small molecule drugs in the pharmaceutical industry. In Figures 2a–c,
separations of common over-the-counter (OTC) analgesic drugs are demonstrated based on
methods described in the United States Pharmacopeia-National Formulary (USP-NF) [20–
22]. These techniques were not directly adapted from previously published monographs
using guidelines established in USP Chapter 621 [23,24], but rather developed using faster
gradient LC approaches that are more common in the pharmaceutical industry for method
screening. Ongoing investigations are focused on determining the full linear range of these
separations and determining the applicability of the technique at trace impurity levels
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(0.05% of the active pharmaceutical ingredient, or API). In general, on-column detection

provides shorter path-lengths than typical flow cells, which can decrease sensitivity;
however, this approach was used to avoid significant extra-column band broadening.

Beyond standard API and impurity testing, dissolution and process monitoring studies are
also critical aspects of pharmaceutical analysis. By coupling the capillary LC platform
directly to a solution vessel, these types of measurements can be made on-line, which
eliminates the need for manual sampling at various time intervals. This is demonstrated in
Figure 2d, which shows a recirculating liquid-handling pump coupled to the LC injector that
took samples intermittently from a stirred vial containing an analgesic tablet composed of
acetaminophen, aspirin, and caffeine. As the tablet slowly dissolved over a period of 11
hours, the peak areas of the components grew as expected (Figures 2e and 2f). Over the full
series of 50 chromatograms in this study, an overall retention time range of 3 s over the
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course of the experiment was observed for the caffeine peak, with an overall relative
standard deviation for retention time under 1%. Such dissolution testing often requires
manual or on-line sampling of larger volumes when traditional analytical-scale LC columns
are used. In this capillary LC example, the relatively large number of injections (50) required
2 μL of sample over the course of the experiment (40 nL per run); this total volume is less
than 10% of the volume of a single injection using sequential injection chromatography for a
similar off-line experiment [25]. Thus, even for experiments conducted over longer time
periods, the overall sample removed can be minimal. In the future, novel sampling
approaches can be devised to provide improved coupling to process streams for on-line
process monitoring using the portable capillary LC instrument.

2.3 Use of the Compact Capillary LC Platform for Forensic Separations

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In addition to the advantage of a smaller footprint in laboratory settings for direct sampling
from an adjacent vessel, another advantage of a portable LC system with an internal battery
supply is that it can be operated remotely and utilized for field applications. This capability
would be especially advantageous for the analysis of illicit drug compounds, both for on-site
seized drug identification and point-of-care screening for substance abuse in clinical
settings. Figure 3 depicts six relevant drug panels, including benzodiazepines, cannabinoids,
stimulants (cocaine and metabolites), and opioids and opioid-related compounds. One

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challenge for this application area is the vast array of molar absorptivity values that exist for
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different drug classes, which can lead to lower sensitivity for compounds that do not absorb
strongly at a given LED wavelength, for example as shown in Figures 3e and 3f in which
lower signals are obtained even though the chemical concentrations are much higher than
several of the other analytes shown. As previously reported, one solution to this issue is to
combine multiple on-column LED-UV detectors that emit light at different wavelengths to
provide absorbance values with higher signals for specific compounds [13]. Thus far, 255
nm and 275 nm have primarily been used, although other LED wavelengths can also be
considered [26].

When combining multiple LED-UV detectors in tandem at the column outlet in the format
described above, an additional advantage is improved identification of target analytes
through the use of dual-wavelength absorbance ratios for the detected compounds [27–30].
In this arrangement, both concentration and path length for each separated compound are
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identical at the adjacent detection points, but the molar absorptivity coefficients are different,
depending on the wavelength of each LED. A ratio of two absorptivity coefficients can
provide an analyte signature that provides more reliable identification than just retention
time. Early results for a 4-compound test mixture using a prototype version of the detector
with dual-wavelength absorbances at 255 nm and 275 nm were recently demonstrated [13];
continuing work is focused on using this approach with other wavelengths for the drug
compounds described in this study.

3. Conclusions
The use of an integrated, portable capillary LC instrument for the analysis of pharmaceutical
and illicit drug compounds was demonstrated for the first time. As shown in the data
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presented, applications that employ LC-UV methodology can be transferred to the new
platform with flow rates that are 100–1,000 times lower than analytical-scale columns while
maintaining similar separation performance. This “greener” approach to LC separations
significantly reduces chemical usage and waste, a key driver toward more sustainable
analytical techniques [31]. As this technology matures, it is likely that its use will expand for
drug analysis and into other application areas that utilize LC-UV, and will be especially
useful in field applications such as environmental testing. Work is ongoing to further
develop methods utilizing this platform, as well as to further expand its capabilities into
other chromatographic modes for even wider use.

Supplementary Material
Refer to Web version on PubMed Central for supplementary material.
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Acknowledgements
Funding for this project was provided by the National Institutes of Health through award R41 DA045382. The
content is solely the responsibility of the authors and does not necessarily represent the official views of the
National Institutes of Health. Ray West and Greg Ward (Axcend Corporation) are acknowledged for technical
assistance. Glenn Kresge and Alexis Zimmer (Rowan University) are acknowledged for helpful conversations
related to this work.

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Figure 1.
3D diagram of the complete portable LC instrument with removable capillary column
cartridge is shown in panel (A). An internal drawing depicting various essential components
in the system is shown in panel (B). The fluidic connections that are formed when the
cartridge is inserted into the instrument and torque is applied are highlighted in panel (C).
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Figure 2.
Pharmaceutical applications of the portable LC instrument. Analytical methods for over-the-
counter analgesic drugs naproxen, ibuprofen, and acetylsalicylic acid (i.e. aspirin) are shown
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in panels (A-C). Panel (D) depicts the use of a M50 pump to circulate a tablet dissolution
vessel through the injection port for on-line repeated analysis (general flow path shown with
white dashed arrows). Selected chromatograms after 10 runs (black trace), 20 runs (blue
trace), 30 runs (green trace), 40 runs (red trace), and 50 runs (orange trace) are shown in
panel (E), with acetaminophen, caffeine, and aspirin peaks resulting from tablet dissolution.
In panel (F), the calculated area for the caffeine peak is shown for all 50 runs (~13 min cycle
time per run). Further experimental details can be found in the Supporting Information.
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Figure 3.
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Chromatograms for various analytical panels of illicit drugs. Panel (A), benzodiazepines: (1)
bromazepam, (2) nitrazepam, (3) diazepam, (4) lorazepam; Panel (B), cannabinoids: (5)
cannabidiolic acid, (6) cannabidiol, (7) cannabinol, (8) Δ9-tetrahydrocannabinol, (9) Δ9-
tetrahydrocannabinolic acid; Panel (C), methadone and metabolite: (10) 2-ethylidene-1,5-
dimethyl-3,3-diphenylpyrrolidine (EDDP) perchlorate, (11) methadone; Panel (D), cocaine
and metabolites: (12) benzoylecgonine, (13) cocaine, (14) cocaethylene; Panel (E), opioid
drugs: (15) oxycodone, (16) hydrocodone, (17) codeine; Panel (F), heroin and metabolites:
(18) morphine, (19) 6-acetylmorphine, (20) heroin. Further experimental details can be
found in the Supporting Information.
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