Accepted Manuscript

Current and future impact of 3D printing on the separation sciences

Umme Kalsoom, Pavel N. Nesterenko, Brett Paull

PII: S0165-9936(18)30008-6
DOI: 10.1016/j.trac.2018.06.006
Reference: TRAC 15172

To appear in: Trends in Analytical Chemistry

Received Date: 9 January 2018

Revised Date: 12 June 2018
Accepted Date: 12 June 2018

Please cite this article as: U. Kalsoom, P.N. Nesterenko, B. Paull, Current and future impact
of 3D printing on the separation sciences, Trends in Analytical Chemistry (2018), doi: 10.1016/
j.trac.2018.06.006.

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 ACCEPTED MANUSCRIPT

1 Current and future impact of 3D printing on the separation sciences

2 Umme Kalsoom,a,b Pavel N. Nesterenko,a,b and Brett Paull a, b,*
(a)
3 Australian Centre for Research on Separation Science (ACROSS), School of Natural Sciences,
4 College of Science and Technology, University of Tasmania, Hobart, 7001, Australia.bret
(b)
5 ARC Centre of Excellence for Electromaterials Science (ACES), School of Natural Sciences, College

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6 of Science and Technology, University of Tasmania, Hobart, 7001, Australia.

7 Corresponding author: Brett Paull

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8 E-mail: brett.paull@utas.edu.au; Fax: +61 3 6226 2858; Tel: +61 3 6226 6680

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10 Abstract

11 The potential of 3D printing to transform the field of separation science is becoming clear, based
12 upon an increasing capacity to create highly customised devices, materials and structures, with

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13 complex geometries. The constantly improving print resolution and increasing variety of available
14 print materials, including functional and composite materials, mean devices can be printed today,
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15 which would be extremely challenging to achieve using traditional manufacturing techniques. This
16 review covers the majority of 3D printed devices to-date designed for use within the separation
17 sciences, categorised under application within pre-separation, separation, and post-separation
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18 stages of analysis. It describes the impact 3D printing is having on the field, both current and future,
19 recent achievements and challenges, and improvements required to reach its maximum potential as
20 a transformative technology.
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21 Key words
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22 3D printing; separation science; chromatography; separation devices; analytical sciences.

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24 1. Introduction to 3D printing

25 Additive manufacturing, more commonly known as 3D printing, continues to impact areas of

26 manufacturing and prototyping, through provision of a quick and low cost alternative to the more
27 traditional manufacturing techniques, such as moulding, forming and computer numerical control
28 machining [1]. In 3D printing, 3D models (computer aided design (CAD) drawing) are fabricated
29 layer-by-layer (LBL) until the whole object is complete. In addition to LBL, continuous liquid interface
30 printing (CLIP), and particle replication in non-wetting templates (PRINT) approaches to 3D printing
31 have also been developed [2, 3]. The various technologies for 3D printing include, fused deposition

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32 modelling (FDM) [4], stereolithography (SLA) [5], selective laser melting or sintering (SLM) [6], and
33 inkjet and polyjet printing [7, 8]. The precise principles, advantages, limitations and applications of
34 each of these techniques is beyond the scope of this review, however details of these and similar

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35 technologies can be readily found elsewhere [8]. In addition, a detailed review on the various
36 materials available for use with 3D printers, their properties, advantages and disadvantages has also
37 recently been published [9].

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38 Since its introduction in the early 1980s [10], 3D printing has seen widespread application within a
39 range of areas, including the environmental sciences, biological sciences, medical device

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40 development, lab-on-a-chip technology, and many other varied fields of manufacturing and
41 engineering. This widespread application is based upon the capacity to rapidly fabricate almost any
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42 complex 3D structure or design, with relatively low cost and comparative ease. 3D printing has
43 already emerged as an impactful fabrication tool for use within the wider analytical sciences. There
44 are several recent reviews describing the application of 3D printing within this field, included those
45 specifically dedicated to the area of microfluidics [8, 11-13]. However, the latest large review by
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46 Spence et al. which focused upon the wider analytical sciences barely mentioned the separation
47 sciences [8]. Despite this, there is absolutely no doubt that the separation sciences currently are, and
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48 will be, significantly impacted by this technology, as already evidenced by a significant number of
49 publications in this area. Figure 1. shows the growing number of publications based upon devices
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50 and materials produced using 3D printing technology and applied in one or more modes of
51 separation science. Therefore, this timely and critical review will discuss this impact, and the future
52 prospect for 3D printing as a supporting technology for the production and development of
53 separation devices, focusing on devices/instruments for application in pre-separation processes,
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54 analytical separations, and post-separation phases of analysis. Furthermore, the advantages and
55 limitations of current 3D printing technology, and what can be done to maximise its future impact in
56 this area, will be discussed. For rapid reference, and to compliment Figure 1, Table 1 is provided as a
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57 summary of the 3D printed separation devices reported to-date, some advantages and/or obvious
58 limitations, and real or potential applications presented.
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59 2. Pre-separation
60 2.1. Sample injection and preparation devices

61 Sample preparation is a crucial step in most analyses, that typically involves one or more processes,
62 such as dilution, filtration, extraction, pre-concentration, purification, or derivatisation, prior to
63 separation and detection, typically to improve the selectivity and sensitivity of the method [14].
64 Over the past few years, there has been an increasing number of reports on the use of 3D printing to
65 produce sample injection and preparation devices for use within, or coupled to, separation methods.
66 For example, these have included injection systems [15], a 3D printed distillation column for
67 purification [16], a 3D printed extraction chamber [17], a 3D printed probe for ambient sampling
68 with low temperature plasma ionisation mass spectrometer (MS) analysis [18, 19], 3D printed solid

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69 phase extraction (SPE) sorbents and devices [15, 20-25], customised sample holders and trays for
70 HPLC-MS [26], and various holders for hyphenation of a sample preparation unit with coupled
71 separation instruments [27].

72 Clearly, the capacity of 3D printing to facilitate the realisation of customised complex fluidic
73 architectures, with a wide range of pre-print or post-print functional (and functionable) materials, is
74 an exciting prospect for the development of highly selective novel devices for sample extraction and
75 concentration. An excellent recent example of this is the development of a unique SPE pre-
76 concentrator cartridge, printed using a standard poly(acrylate) resin, without any additional

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77 modification, for the selective extraction of trace metal ions from seawater. Using 3D printing it was
78 possible to produce an ordered porous structure, based upon cuboid entities, distributed
79 throughout an extraction channel [20]. The novel design of the device included an extraction phase

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80 with a sufficient surface area and offered improved fluid mixing, which prevented blockage of the
81 flow channels between the cuboids (See Fig. 2). However, the limited printer resolution in this study
82 prevented the fabrication of spherical structures and restricted the designs to cuboids with a

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83 minimum print size of 0.4 mm × 0.4 mm × 0.2 mm (length × width × height). The extraction of metal
84 ions (Mn, Ni, Cu, Zn, Cd, Pb) from natural waters at pH 8 was performed utilising the ion-exchange
85 and complexing properties of carboxylic groups on the surface of the printed porous structure [20].

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86 The total surface area of the cuboids in the pre-concentrator was 4.9 cm2, with an adsorption
87 capacity of 0.14 µmol cm-2, providing recovery of each of the metal ions studied in the range 83 -
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88 109% from 1 mL samples. In a similar report, lipophilic interactions between a printed sorbent
89 material and small drug molecules (glimepride) were exploited, in this case within a solid phase
90 micro-extraction (SPME) based method. Again, no pre- or post-print chemical modification of the
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91 material was carried out, and the printed sorbent applied to extraction from waters samples [21].

92 In addition to direct 3D printing, the use of print technologies to produce substrates for
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93 immobilisation of selective sorbent particles has also been reported [25]. In a most recent example
94 of this, Middeleer et al., reported the immobilisation of molecularly imprinted polymers (MIPs), via
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95 polymer network crosslinking onto 3D printed poly-ε-caprolactone (PCL) scaffolds, and applied these
96 composite substrates to the SPE of small molecules [25]. Further to surface modification, 3D printing
97 also provides the opportunity for incorporation of functional materials e.g. nano-particles, into the
98 base polymer prior to printing, thus facilitating the fabrication of more selective extraction sorbents.
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99 For example, recently Su and Chen incorporated TiO2 nanoparticles into a print material to produce a
100 stacked cuboid composite sorbent. 3D printed column holders were also produced, containing the
101 fittings for connectors, within which the 3D printed composite packing was inserted and
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102 subsequently employed for sample pre-treatment, and the selective extraction of As and Se species
103 from water and urine samples, prior to ICP-MS analysis [28].
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104 A number of integrated 3D flow based systems have been reported to-date, which demonstrate the
105 significant potential of 3D printing technology in the move towards fully automated multifunctional
106 analytical systems. Calderilla et al. recently reported on the use of SLA 3D printing to fabricate a two-
107 piece modular device for the flow injection analysis (FIA) based determination of iron, where the
108 first unit contained a pre-SPE mixer/reactor (12.2 × 2.1 mm, L x W), and the second unit included a
109 SPE chelating disk support (truncated cone 1 × 7 × 3.7 mm, smaller I.D. × larger I.D. × H) and a post-
110 column reactor (same size as pre-SPE mixer). Both units had integrated connectors. These units were
111 connected to accommodate the SPE disk between them, containing 8 µm particles of an
112 iminodiacetic acid functionalised poly(styrene-divinylbenzene) resin (Empore Chelating disk,
113 Empore, USA). Sample oxidation with H2O2 to convert iron(II) to iron(III) prior to SPE for the
114 determination of total iron was performed in the first unit, and iron(III) pre-concentration on the

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115 chelating disk, elution and post-column reaction (PCR) with ammonium thiocyanate, prior to UV
116 detection, was conducted in the second unit [23]. This apparently robust 3D printed system was
117 used for the automated speciation of Fe using a multi-syringe FIA system. Integration of the 3D
118 printed devices within the automated FIA technique provided improved sensitivity, with an 85%
119 increase in peak height reported. The complete set-up containing the sorbent disk for the
120 automated speciation of iron is shown in Fig. 3. The same approach with minor modifications was
121 used later by the same group for the pre-concentration and analysis of Cr(VI) in water samples [24].
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123 In a similar study, SLA 3D printing was used for production of a FIA system, which included a column

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124 packed with TrisKem Pb (PB-B25-S, 50–100 μm, TrisKem, France), a neutral moderate polarity
125 poly(acrylate) resin coated with 4,4’(5’)-di-t-butylcyclohexano-18-crown-6, and developed for the
126 SPE of lead. A mixing coil for post-column reaction with 4-(2-pyridylazo)resorcinol (PAR) and a flow
127 cell for visible detection were also included. This printed platform was applied to the on-line

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128 determination of Pb in river water samples [29]. A 3D printed µFIA system with an integrated µSPE
129 chip and a flow injection device has also been reported, in this case developed for automatic
130 sampling within an online HPLC based method [15].

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131
132 Based on the above studies it can be observed that SLA has been the favoured printing technique
133 employed for the fabrication of flow systems consisting of small I.D. tubing/ mixing coils, which are

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134 difficult to produce using alternative 3D printers, due to surface roughness, blocked channels and
135 generally poorer print resolution. Mattio et al. recently reported upon the clogging of the narrow
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136 channels (mixing coil of a flow system) produced using polyjet printing, due to incomplete removal
137 of water soluble support material. The same system was printed using SLA without clogging,
138 however, an uneven surface and the presence of micro-cracks from shrinkage and deformation was
139 observed, a common issue with SLA, which resulted in leaking of the reagents and poor detection
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140 (due to deformation of optical path length). Digital light processing (DLP) was found to be best
141 option for printing the flow system with mixing coil, however, polyjet printing showed better
142 performance than DLP for manufacturing of the flow system without the coil [30].
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144 2.2. Reactors
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145 3D printing is establishing itself as a highly versatile manufacturing tool for the development of
146 bespoke fluidic devices, which can be utilised with commercial flow systems, and/or spectroscopic
147 and chromatographic instruments, to facilitate online analysis and optimisation of chemical
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148 reactions. Exploiting the design flexibility of 3D printing, Capel et al. were able to manufacture tailor-
149 made reactors using SLA, which could be coupled to existing laboratory instruments, specifically
150 HPLC. However, poor thermal conductivity of the poly(acrylate) based printing material used in
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151 initial studies resulted in non-uniform temperature distribution and excessive delays from heating
152 and cooling of the reactors. To overcome this limitation, stainless steel reactors capable of working
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153 at elevated temperatures and pressures have recently been fabricated using SLM. For example, a
154 multifunctional thermally compatible flow reactor in stainless steel was fabricated using SLM and
155 coupled to an HPLC instrument, to develop a fully automated flow system capable of performing the
156 entire reaction, analysis and optimisation [31]. Gutmann et al. also manufactured a stainless steel
157 flow reactor with similarly high thermal, chemical and mechanical resistance [32]. Other examples of
158 such work include 3D printed reaction vessels [33], reaction cells for online monitoring using MS [34,
159 35] and fluorescence detection [36], reaction-ware for purification [37], an interface to integrate the
160 enzyme reactors with LC-MS [38], and a platform for enzymatic isolation prior to identification using
161 LC-MS [39]. These made-to-order reaction devices (RD) provide new opportunities for system
162 automation and therefore minimise sample handling, and generally simplify equipment
163 requirements.

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164 3. Separation
165 3.1. Columns and stationary phases

166 The ability of 3D printing technique to produce complex tailor-made designs with greater control
167 and precision (within a 3D space), makes it a technique of choice for developing miniaturised
168 analytical instruments and potentially integrated chromatography platforms. Specifically,
169 miniaturisation of chromatographic systems, including the production of long separation columns
170 within smaller coiled footprints, can be realised via 3D printing. For example, Lucklum et al. used SLA
171 based printing to develop miniature planar, and stacked spiral 3D GC columns, for application within

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172 a portable ethylene sensor system. Using a high resolution 3D printer, they were able to design 45
173 cm long stacked spiral columns with small footprints, and highly precise round cross-sections. The
174 researchers’ claimed the printed columns provided advantages in relation to improved packing

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175 density for the stationary phase, and consequently an improved baseline resolution of the peaks in a
176 relatively short run time (∼15 min), in comparison to a 50 cm long planar silicon column with
177 rectangular geometry [40]. Additionally, the integration of previously separate components (e.g. gas-

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178 tight connectors) simplified the instrument design and prevented any connection issues typically
179 observed with portable instruments. The same group also developed a GC system, with a 3D printed
180 column, that was used for the sensitive (ppb level) detection of ethylene gas [41]. Poly(acrylate) was

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181 employed for this work, though the experiments were conducted at a maximum temperature of 65
182 °C, as all polymeric printing materials generally exhibit limited thermal stability [9, 42, 43], which
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183 make them unsuitable for typical temperature conditions used for GC.

184 The capacity of 3D printing to produce tailor-made geometrically complex and defined materials,
185 provides new opportunities for the optimisation of column structures and stationary phases, which
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186 would be difficult, if not impossible, to achieve using traditional manufacturing techniques. For
187 example, Fee and co-workers utilised the capability of 3D printing technology to design porous
188 columns with highly precise internal pore geometries, which provided great control over packing
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189 morphology and structure. The freedom to orientate individual ‘units’ (representing a typical
190 stationary phased particle) within these printed porous beds, provided opportunities for the
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191 production of stationary phase beds centred upon stationary phase units other than typical spheres,
192 with homogenous porosity all through the bed. Three phase geometries, including simple cubic
193 arrangement (SC), parallel channels (PC) and herringbone channels (HC) (Fig. 4) were printed and
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194 characterised. The shape of the packing units was restricted to polyhedrons and polygons (from
195 software limitations), in comparison to typical spheres. The one step fabrication capability of 3D
196 printing allowed the fabrication of the entire integrated column, including internal flow distributers,
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197 external connectors and packing, which provided increased structural robustness [44]. Further
198 studies by this group, focused on the comparison of theoretical studies/predictions vs experimental
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199 results for 3D printed columns of different element shapes and packing arrangements. Their
200 investigations showed that the spherical particles, usually considered as the ideal shape for column
201 packing, could be outperformed by other 3D printed particle shapes (e.g. tetrahedral, minimum
202 plate height (mm) = 0.90 ± 0.11), an observation that could possibly affect the nature of next
203 generation packing materials and future chromatography columns [45]. A non-porous polymeric
204 material was selected for printing, however a porous material with finer resolution elements to
205 reduce theoretical plate height was suggested for improving the column performance. Polymeric
206 columns are also prone to degradation under moderate temperature and pressure conditions, and
207 have low organic solvent compatibility, and so future investigations into inorganic print materials for
208 liquid phase separations are also required.

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209 Moving in the direction of printed columns for high pressure LC separations, Sandron et al. used SLM
210 printing technique to produce 60 cm long coiled capillary columns (0.9 mm I.D.) in both stainless
211 steel and titanium alloy, which provided high pressure and thermal stability. These compact columns
212 (external dimensions of 5 x 30 x 58 mm) were tested with two types of stationary phase, first being
213 slurry packed with various sized octadecyl silica microparticles, and secondly via an in-situ prepared
214 poly(butylmethacrylate-ethyleneglycol dimethacrylate) monolith [46]. The printed columns were
215 employed for the separation of a phenone mixture, however, the chromatographic performance of
216 this initial printed column was rather limited (791-3909 plates/meter for various stationary phases)
217 and further optimisation of the design was required. In a subsequent study, columns with improved

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218 design and performance enabled the separation of a mixture of 10 proteins under reversed-phase LC
219 conditions (both column design and separation are shown in Fig. 5). In this work the SLM printer’s
220 resolution limited the channel inner diameter to ~1 mm and the surface roughness of the channel
221 walls was dependent upon the metallic powder used to print (~ 20 um) [47]. In the most recent

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222 report by Gupta et al. the effect of column geometries on the separation efficiency of the 3D printed
223 titanium columns was studied in considerable detail. SLM printing was used for the fabrication of
224 three columns, of equal length and I.D, but with differing geometries, including 2D planar

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225 serpentine, 3D spiral, 3D serpentine, with a low, medium and high number of high aspect ratio turns,
226 respectively. The 3D serpentine column with the highest number of high aspect ratio turns showed
227 better performance in terms of increased plate number (up to 245 % average increase), improved

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228 peak capacity (up to 82 %) and reduced analysis time (58 %), in comparison to other two column
229 geometries [48].
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230
231 Both the above studies on particle and stationary phase bed design/optimisation, and column
232 geometry, respectively, demonstrate how 3D printing can, and is, impacting upon chromatography
233 column design and production for the future. Clearly the next challenge is the production of a
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234 printed phase within a simultaneously printed pressure and solvent compatible column, which could
235 begin to deliver some high performance chromatographic separations.
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237 3.2. Thin layer chromatography

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238 Besides printing of stationary phases and structures for column chromatography, 3D printing has
239 also been used for the fabrication of thin layer chromatography (TLC) plates and stationary phases
240 [49-51]. The ability of 3D printing to deposit precise layers of stationary phase, of different length,
241 width and structure, facilitates the customisation of the TLC plates to the desired application, and
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242 potentially reduces production costs. Improved phase homogeneity may also help improve solvent
243 flow profiles and peak efficiency in the future, problems currently associated with classic TLC plates.
244 For example, Fichou and Morlock deposited the layers of silica gel using a modified commercial 3D
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245 printer, wherein the plastic extruder of the printed was replaced with a slurry doser (which was itself
246 also 3D printed). These TLC plates were employed for the separation and semi-quantitative analysis
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247 of a mixture of dyes [50]. The results obtained were comparable to commercially available TLC
248 plates. More recently Macdonald et al. manufactured a micro-structured poly(acrylate) layer which
249 was used as stationary phase for TLC separations of dyes and proteins without any additional surface
250 modifications [51]. In this case the print included both the support platform (plate) and the
251 stationary phase in a single print, making a low-cost and disposable option for quick and easy TLC
252 separations.

253 The above reports would suggest significant opportunities for the printing of future TLC plates with
254 multiple lanes of stationary phases, perhaps of varying selectivity, onto a single plate (utilising multi-
255 -material printing), which is once again not easily (or cheaply) achievable using current
256 manufacturing techniques.

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257 4. Post-separation
258 4.1. Interfaces

259 Various devices for post-separation steps, including prototypes to introduce samples into various
260 detection systems; interfaces to hyphenate the separation unit with the detection systems; and
261 detector bodies/housings, have all been manufactured using 3D printing in recent years. Salentjin et
262 al. fabricated a finger sized paper spray ionization (PSI) cartridge for extended solvent spray
263 generation through a paper tip (without the need for a pump) using an FDM printer. In paper spray
264 (PS) the samples are deposited and stored on the paper for extraction/ionisation directly from the

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265 dried sample spot. As a proof-of-principle, this device was used for an online paper chromatography-
266 MS experiment for differential retention of dyes (Fig. 6) [52]. The 3D printed cartridges provided the
267 stability and support to the paper tip and facilitated the alignment of the tip with the MS. Integrated

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268 fluid control features allowed fast wetting and continuous solvent supply without the need for
269 external pumps, advantages simply not possible to achieve using traditional milling techniques,
270 which would be time consuming and require careful machining to produce replicates with high

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271 precision. In other work from the same group, 3D printing was employed to re-design a customised
272 PS cartridge with integrated SPE, which was initially produced by milling, for the selective and
273 sensitive analysis of plasma samples for target drug molecules. This reproducible 3D printed device

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274 was produced in much less time (approx. 4hrs) in comparison to the traditional milling process,
275 which required almost a week to manufacture sufficient cartridges for batch analysis. Furthermore,
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276 3D printing allowed the fabrication of a highly customised, inexpensive (US$2) device with complex
277 parts, and of smaller size compared to its prototype, that would be very challenging to produce using
278 traditional processing [53].
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279 4.2. Detectors/ detection housing

280 Detection techniques including UV/Vis absorption, electrochemical detection, fluorescence or

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281 chemiluminescence are commonly used to record the separation provided from the separation
282 platform (chromatographic or electrophoretic). Traditional techniques for detector manufacturing
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283 rely on removing material from a block until the desired structure is obtained e.g. computer
284 numerical control (CNC) routing [54]. However, using these techniques, it is hard to produce
285 complex and hollow structures within a single object. With the availability of 3D printing, many
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286 groups have now begun producing and optimising their own detector cells. For example, using FDM
287 printing, Prikryl and Foret designed a horn shape hole in the fluorescence detection head, which
288 would be very expensive and difficult to produce from standard milling/machining process. This
289 detector was used for the detection of fluorescently tagged oligosaccharides separated by CE [55].
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290 Further application of this detection system in building a simple CE system with in-built
291 minicomputer for image based data acquisition have also been explored [56]. Similarly, a single
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292 piece detection body accommodating the detection components (e.g. LED, photodiode) and a
293 hollow slit was recently manufactured using an FDM 3D printer. The impact of various spatial
294 orientations on print resolution was studied to allow the printing of a hollow slit as narrow as 70 µm.
295 This detector body can be used to perform optical detection in CE, LC and FIA systems (Fig. 7) [54].

296 Geometrically complex 3D printed flow cells for chemiluminescence detection have also recently
297 been reported [57, 58]. Spilstead et al. compared the performance of 3D printed and CNC milled
298 flow cells for chemiluminescence detection within FIA manifolds. Their results demonstrated that
299 their tailored 3D printed flow cell provided improved detection over traditional coiled tubing based
300 cells, and provided advantages such as enhanced transfer of the emitted light from the translucent
301 white printing material, and reproducible positioning of the photomultiplier on the detection zone
302 [57]. Gupta et al. applied a 3D printed chemiluminescent flow cell based upon a novel radial design

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303 within both FIA and ion chromatography [58]. Once again the novel 3D printed design, which
304 provided improved performance over alternative designs, would be difficult to produce using
305 traditional manufacturing.

306 Other recent examples of 3D printed detection systems/bodies include; a 3D printed stand-alone ion
307 mobility spectrometer developed for the identification and detection of organic compounds [59]; in-
308 house fabrication of mm-wave sensor for LC [60], 3D printed wall-jet electrode device for
309 electrochemical detection in HPLC [61], and a custom made 3D printed holder for mounting the
310 optical fibre to a fluorimeter to facilitate fluorescence detection using HPLC [62]. Also, the use of 3D

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311 printing for visualisation of reaction progress surfaces from NMR, IR and HPLC raw data has also
312 been described [63] which shows the potential of this technology to be used as a useful teaching
313 tool.

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314 5. Conclusion, challenges and future direction

315 3D printing is becoming an increasingly popular production technology for use within the separation

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316 sciences, with new printed devices for pre-separation, separation, and post-separation increasingly
317 being reported. The recent trend for fully automated miniaturised instruments is increasing the
318 popularity of this rapid and versatile fabrication technology. The possibility of print-to-order

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319 production of columns and stationary phases, and the continuing development of new printing
320 materials, could speed the development of highly customised, sensitive and selective separation
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321 platforms. Furthermore, the availability of multi-material printers provides the option for production
322 of various components such as columns, stationary phases, flow connectors etc. using materials with
323 variable properties (e.g. porosity, functionality, thermal resistance etc.).
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324 However, despite the rapid advances in this technology to-date, there are some constraints to
325 obtaining the above goals. Currently the low resolution of most of the cost-effective 3D printers can
326 result in surface roughness due to deposition of materials on the surface/ uncured particles, and an
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327 inability to produce micro-scale structures. The print resolution currently available in affordable
328 printers is at best an order of magnitude away from that required to print viable stationary beds for
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329 actual ‘high-performance’ separations, or indeed high capacity SPE phases. However, the technology
330 is developing at an impressive rate, and continuous improvements in the resolution of commercial
331 3D printers, and an ever increasing variety of printing materials, will help overcome current
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332 limitations in the very near future.

333 6. References
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334 1. Z.-X. Low, Y.T. Chua, B.M. Ray, D. Mattia, I.S. Metcalfe, D.A. Patterson, Perspective on 3D
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335 printing of separation membranes and comparison to related unconventional fabrication

336 techniques, J. Membr. Sci. 523 (2017) 596-613.
337 2. S.W. Morton, K.P. Herlihy, K.E. Shopsowitz, Z.J. Deng, K.S. Chu, C.J. Bowerman, J.M.
338 DeSimone, P.T. Hammond, Scalable manufacture of built-to-order nanomedicine: spray-
339 assisted layer-by-layer functionalization of PRINT nanoparticles, Adv. Mater. 25 (2013) 4707-
340 4713.
341 3. J.R. Tumbleston, D. Shirvanyants, N. Ermoshkin, R. Janusziewicz, A.R. Johnson, D. Kelly, K.
342 Chen, R. Pinschmidt, J.P. Rolland, A. Ermoshkin, E.T. Samulski, J.M. DeSimone, Continuous
343 liquid interface production of 3D objects, Science 347 (2015) 1349-1352.
344 4. I. Zein, D.W. Hutmacher, K.C. Tan, S.H. Teoh, Fused deposition modeling of novel scaffold
345 architectures for tissue engineering applications, Biomaterials 23 (2002) 1169-1185.

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346 5. A.K. Au, W. Lee, A. Folch, Mail-order microfluidics: evaluation of stereolithography for the
347 production of microfluidic devices, Lab. Chip. 14 (2014) 1294-1301.
348 6. M. Agarwala, D. Bourell, J. Beaman, H. Marcus, J. Barlow, Direct selective laser sintering of
349 metals, Rapid Prototyp. J. 1 (1995) 26-36.
350 7. S.F.S. Shirazi, S. Gharehkhani, M. Mehrali, H. Yarmand, H.S.C. Metselaar, N.A. Kadri, N.A. Abu
351 Osman, A review on powder-based additive manufacturing for tissue engineering: selective
352 laser sintering and inkjet 3D printing, Sci. Technol. Adv. Mater. 16 (2015).
353 8. B. Gross, S.Y. Lockwood, D.M. Spence, Recent advances in analytical chemistry by 3D
354 printing, Anal. Chem. 89 (2017) 57-70.

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355 9. S.C. Ligon, R. Liska, J. Stampfl, M. Gurr, R. Mülhaupt, Polymers for 3D Printing and
356 customized additive manufacturing, Chem. Rev. 117 (2017) 10212-10290.
357 10. C.W. Hull, Apparatus for production of three-dimensional objects by stereolithography. 1986,
358 Google Patents, US4575330 A.

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359 11. C. Chen, B.T. Mehl, A.S. Munshi, A.D. Townsend, D.M. Spence, R.S. Martin, 3D-printed
360 microfluidic devices: fabrication, advantages and limitations-a mini review, Anal. Methods 8
361 (2016) 6005-6012.

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362 12. R. Amin, S. Knowlton, A. Hart, B. Yenilmez, F. Ghaderinezhad, S. Katebifar, M. Messina, A.
363 Khademhosseini, S. Tasoglu, 3D-printed microfluidic devices, Biofabrication 8 (2016) 022001.
364 13. M.T.W. Hearn, Trends in additive manufacturing of chromatographic and membrane

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365 materials, Curr. Opin. Chem. Eng 18 (2017) 90-98.
366 14. S. de Koning, H.-G. Janssen, U.A.T. Brinkman, Modern methods of sample preparation for GC
AN
367 analysis, Chromatographia 69 (2009) 33.
368 15. H. Wang, D.J. Cocovi-Solberg, B. Hu, M. Miró, 3D-printed microflow injection analysis
369 platform for online magnetic nanoparticle sorptive extraction of antimicrobials in biological
370 specimens as a front end to liquid chromatographic assays, Anal. Chem. 89 (2017) 12541–
M

371 12549.
372 16. S. Mardani, L.S. Ojala, P. Uusi-Kyyny, V. Alopaeus, Development of a unique modular
373 distillation column using 3D printing, Chem. Eng. Process. 109 (2016) 136-148.
D

374 17. C. Worawit, D.J. Cocovi-Solberg, P. Varanusupakul, M. Miró, In-line carbon nanofiber
375 reinforced hollow fiber-mediated liquid phase microextraction using a 3D printed extraction
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376 platform as a front end to liquid chromatography for automatic sample preparation and
377 analysis: A proof of concept study, Talanta 185 (2018) 611-619.
378 18. S. Martinez-Jarquin, A. Moreno-Pedraza, D. Cazarez-Garcia, R. Winkler, Automated chemical
379 fingerprinting of Mexican spirits derived from Agave (tequila and mezcal) using direct-
EP

380 injection electrospray ionisation (DIESI) and low-temperature plasma (LTP) mass
381 spectrometry, Anal. Methods 9 (2017) 5023-5028.
382 19. S. Martinez-Jarquin, A. Moreno-Pedraza, H. Guillen-Alonso, R. Winkler, Template for 3D
C

383 printing a low-temperature plasma probe, Anal. Chem. 88 (2016) 6976-6980.

384 20. C.-K. Su, P.-J. Peng, Y.-C. Sun, Fully 3D-printed preconcentrator for selective extraction of
AC

385 trace elements in seawater, Anal. Chem. 87 (2015) 6945-6950.

386 21. M. Belka, S. Ulenberg, T. Bączek, Fused deposition modeling enables the low-cost fabrication
387 of porous, customized-shape sorbents for small-molecule extraction, Anal. Chem. 89 (2017)
388 4373-4376.
389 22. L. Konieczna, M. Belka, M. Okońska, M. Pyszka, T. Bączek, New 3D-printed sorbent for
390 extraction of steroids from human plasma preceding LC–MS analysis, J. Chromatogr. A 1545
391 (2018) 1-11.
392 23. C. Calderilla, F. Maya, V. Cerdà, L.O. Leal, 3D printed device including disk-based solid-phase
393 extraction for the automated speciation of iron using the multisyringe flow injection analysis
394 technique, Talanta 175 (2017) 463-469.
395 24. C. Calderilla, F. Maya, V. Cerdà, L.O. Leal, 3D printed device for the automated
396 preconcentration and determination of chromium (VI), Talanta 184 (2018) 15-22.

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397 25. G. De Middeleer, P. Dubruel, S. De Saeger, Molecularly imprinted polymers immobilized on
398 3D printed scaffolds as novel solid phase extraction sorbent for metergoline, Anal. Chim.
399 Acta 986 (2017) 57-70.
400 26. V. Kertesz, T.M. Weiskittel, G.J. Van Berkel, An enhanced droplet-based liquid microjunction
401 surface sampling system coupled with HPLC-ESI-MS/MS for spatially resolved analysis, Anal.
402 Bioanal. Chem 407 (2015) 2117-2125.
403 27. Y.R. Tang, H.H. Yang, P.L. Urban, Prototype of an interface for hyphenating distillation with
404 gas chromatography and mass spectrometry, Mass Spectrom. (Tokyo) 6 (2017) S0061.
405 28. C.-K. Su, W.-C. Chen, 3D-printed, TiO2 NP–incorporated minicolumn coupled with ICP-MS for

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406 speciation of inorganic arsenic and selenium in high-salt-content samples, Microchimica Acta
407 185 (2018) 268.
408 29. E. Mattio, F. Robert-Peillard, C. Branger, K. Puzio, A. Margaillan, C. Brach-Papa, J. Knoery, J.-
409 L. Boudenne, B. Coulomb, 3D-printed flow system for determination of lead in natural

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410 waters, Talanta 168 (2017) 298-302.
411 30. E. Mattio, F. Robert-Peillard, L. Vassalo, C. Branger, A. Margaillan, C. Brach-Papa, J. Knoery,
412 J.-L. Boudenne, B. Coulomb, 3D-printed lab-on-valve for fluorescent determination of

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413 cadmium and lead in water, Talanta 183 (2018) 201-208.
414 31. A.J. Capel, A. Wright, M.J. Harding, G.W. Weaver, Y.Q. Li, R.A. Harris, S. Edmondson, R.D.
415 Goodridge, S.D.R. Christie, 3D printed fluidics with embedded analytic functionality for

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416 automated reaction optimisation, Beilstein J. Org. Chem. 13 (2017) 111-119.
417 32. B. Gutmann, M. Kockinger, G. Glotz, T. Ciaglia, E. Slama, M. Zadravec, S. Pfanner, M.C. Maier,
AN
418 H. Gruber-Wolfler, C. Oliver Kappe, Design and 3D printing of a stainless steel reactor for
419 continuous difluoromethylations using fluoroform, React. Chem. Eng. 2 (2017) 919-927.
420 33. F. Lederle, C. Kaldun, J.C. Namyslo, E.G. Hubner, 3D-printing inside the glovebox: A versatile
421 tool for inert-gas chemistry combined with spectroscopy, Helv. Chim. Acta 99 (2016) 255-
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422 266.
423 34. J.S. Mathieson, M.H. Rosnes, V. Sans, P.J. Kitson, L. Cronin, Continuous parallel ESI-MS
424 analysis of reactions carried out in a bespoke 3D printed device, Beilstein J. Nanotechnol. 4
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425 (2013) 285-291.

426 35. G. Scotti, S.M.E. Nilsson, M. Haapala, P. Poho, G.B.A. Gennas, J. Yli-Kauhaluoma, T. Kotiaho,
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427 A miniaturised 3D printed polypropylene reactor for online reaction analysis by mass
428 spectrometry (vol 2, pg 299, 2017), React. Chem. Eng. 2 (2017) 811-811.
429 36. C.K. Su, S.C. Yen, T.W. Li, Y.C. Sun, Enzyme-immobilized 3D-printed reactors for online
430 monitoring of rat brain extracellular glucose and lactate, Anal. Chem. 88 (2016) 6265-6273.
EP

431 37. P.J. Kitson, M.D. Symes, V. Dragone, L. Cronin, Combining 3D printing and liquid handling to
432 produce user-friendly reactionware for chemical synthesis and purification, Chem. Sci. 4
433 (2013) 3099-3103.
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434 38. A. Jönsson, R.R. Svejdal, N. Bøgelund, T.T.T.N. Nguyen, H. Flindt, J.P. Kutter, K.D. Rand, J.P.
435 Lafleur, Thiol-ene monolithic pepsin microreactor with a 3D-printed interface for efficient
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436 UPLC-MS peptide mapping analyses, Anal. Chem. 89 (2017) 4573-4580.

437 39. M.-M. Wang, P. Laborda, L.P. Conway, X.-C. Duan, K. Huang, L. Liu, J. Voglmeir, An integrated
438 3D-printed platform for the automated isolation of N-glycans, Carbohydr. Res. 433 (2016)
439 14-17.
440 40. F. Lucklum, S. Janssen, W. Lang, M.J. Vellekoop, Miniature 3D gas chromatography columns
441 with integrated fluidic connectors using high-resolution stereolithography fabrication,
442 Procedia Eng. 120 (2015) 703-706.
443 41. N.A. Zaidi, M.W. Tahir, M.J. Vellekoop, W. Lang, A gas chromatographic system for the
444 detection of ethylene gas using ambient air as a carrier gas, Sensors 17 (2017) 2283.
445 42. U. Kalsoom, A. Peristyy, P.N. Nesterenko, B. Paull, A 3D printable diamond polymer
446 composite: a novel material for fabrication of low cost thermally conducting devices, RSC
447 Adv. 6 (2016) 38140-38147.

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 ACCEPTED MANUSCRIPT
448 43. U. Kalsoom, P.N. Nesterenko, B. Paull, Recent developments in 3D printable composite
449 materials, RSC Advances 6 (2016) 60355-60371.
450 44. C. Fee, S. Nawada, S. Dimartino, 3D printed porous media columns with fine control of
451 column packing morphology, J. Chromatogr. A 1333 (2014) 18-24.
452 45. S. Nawada, S. Dimartino, C. Fee, Dispersion behavior of 3D-printed columns with
453 homogeneous microstructures comprising differing element shapes, Chem. Eng. Sci. 164
454 (2017) 90-98.
455 46. S. Sandron, B. Heery, V. Gupta, D.A. Collins, E.P. Nesterenko, P.N. Nesterenko, M. Talebi, S.
456 Beirne, F. Thompson, G.G. Wallace, D. Brabazon, F. Regan, B. Paull, 3D printed metal

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457 columns for capillary liquid chromatography, Analyst 139 (2014) 6343-6347.
458 47. V. Gupta, M. Talebi, J. Deverell, S. Sandron, P.N. Nesterenko, B. Heery, F. Thompson, S.
459 Beirne, G.G. Wallace, B. Paull, 3D printed titanium micro-bore columns containing polymer
460 monoliths for reversed-phase liquid chromatography, Anal. Chim. Acta 910 (2016) 84-94.

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461 48. V. Gupta, S. Beirne, P.N. Nesterenko, B. Paull, Investigating the Effect of Column Geometry
462 on Separation Efficiency using 3D Printed Liquid Chromatographic Columns Containing
463 Polymer Monolithic Phases, Anal. Chem. 16 (2018) 1186-1194.

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464 49. R. Groarke, T. Skrobisz, C. Hughes, R. McCann, A. Scigliano, D. Brabazon, Next generation
465 high performance thin layer chromatography (HP-TLC) using additively manufactured
466 acrylate based polymer stationary phases, in International Symposium on Chromatography.

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467 2016: University College Cork, Ireland.
468 50. D. Fichou, G.E. Morlock, Open-source-based 3D printing of thin silica gel layers in planar
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469 chromatography, Anal. Chem. 89 (2017) 2116-2122.
470 51. N.P. Macdonald, S.A. Currivan, L. Tedone, B. Paull, Direct production of microstructured
471 surfaces for planar chromatography using 3D printing, Anal. Chem. 89 (2017) 2457-2463.
472 52. G.I.J. Salentijn, H.P. Permentier, E. Verpoorte, 3D-printed paper spray ionization cartridge
M

473 with fast wetting and continuous solvent supply features, Anal. Chem. 86 (2014) 11657-
474 11665.
475 53. C. Zhang, B.J. Bills, N.E. Manicke, Rapid prototyping using 3D printing in bioanalytical
D

476 research, Bioanalysis 9 (2017) 329-331.

477 54. F. Cecil, M. Zhang, R.M. Guijt, A. Henderson, P.N. Nesterenko, B. Paull, M.C. Breadmore, M.
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478 Macka, 3D printed LED based on-capillary detector housing with integrated slit, Anal. Chim.
479 Acta 965 (2017) 131-136.
480 55. J. Prikryl, F. Foret, Fluorescence detector for capillary separations fabricated by 3D printing,
481 Anal. Chem. 86 (2014) 11951-11956.
EP

482 56. M. Szarka, A. Guttman, Smartphone cortex controlled real-time image processing and
483 reprocessing for concentration independent LED induced fluorescence detection in capillary
484 electrophoresis, Anal. Chem. 89 (2017) 10673-10678.
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485 57. K.B. Spilstead, J.J. Learey, E.H. Doeven, G.J. Barbante, S. Mohr, N.W. Barnett, J.M. Terry, R.M.
486 Hall, P.S. Francis, 3D-printed and CNC milled flow-cells for chemiluminescence detection,
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487 Talanta 126 (2014) 110-115.

488 58. V. Gupta, P. Mahbub, P.N. Nesterenko, B. Paull, A new radial flow-cell fabricated by 3D
489 printing and its evaluation for chemiluminescence detection of hydrogen peroxide, Anal.
490 Chim. Acta 1005 (2017) 81-92.
491 59. A. Hollerbach, Z. Baird, R.G. Cooks, Ion separation in air using a three-dimensional printed
492 ion mobility spectrometer, Anal. Chem. 89 (2017) 5058-5065.
493 60. Y. Zhang, S. Declerck, D. Mangelings, G. He, V. Matvejev, Y. Vander Heyden, J. Stiens, A label-
494 free detector for liquid chromatography systems using mm-wave technology: First proof of
495 concept, J. Chromatogr. A 1516 (2017) 79-88.
496 61. A.S. Munshi, R.S. Martin, Microchip-based electrochemical detection using a 3-D printed
497 wall-jet electrode device, Analyst 141 (2016) 862-869.

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498 62. G.G. Siano, M. Montemurro, M.R. Alcaráz, H.C. Goicoechea, Open-source assisted laboratory
499 automation through graphical user interfaces and 3D printers: application to equipment
500 hyphenation for higher-order data generation, Anal. Chem. 89 (2017) 10667-10672.
501 63. C.S. Higman, H. Situ, P. Blacklin, J.E. Hein, Hands-on data analysis: using 3D printing to
502 visualize reaction progress surfaces, J. Chem. Educ. 94 (2017) 1367-1371.
503 64. J.B. Hu, T.R. Chen, C.H. Chang, J.Y. Cheng, Y.C. Chen, P.L. Urban, A compact 3D-printed
504 interface for coupling open digital microchips with Venturi easy ambient sonic-spray
505 ionization mass spectrometry, Analyst 140 (2015) 1495-1501.
506

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Table 1. A summary of 3D printed devices and their applications in separation sciences.
Printed object and its
dimensions (if provided)
Stator (0.8 mm I.D., 8 mm
long connector) for sample
handling, and SPE chip
consisting of a single
channel (0.8 × 3.0 × 50
mm) with integrated
connectors to fluid lines
Distillation column, 11.2 ×
7.9 × 15.1 cm (L × W × H).
Extraction chamber (12 × 3
mm I.D, 7 mm O.D)
Probe for analysis of
samples under ambient
conditions
SPE cartridge with 29 layers Poly(acrylate)
of 526 total cuboids in the
ACCEPTED MANUSCRIPT
Material Printing technique Application Advantages (A)/limitation (L) Analytical
(Printer, producer) mode application, refs
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Pre-separation
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Poly(acrylate) SLA, Form 2, LC A: Automatic flow control of pressure Analysis of 4-
(Formlabs, USA) driven fluid, improved sensitivity hydroxybenzoate
(enhancement factor, 16−25). analogues and
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triclosan in
biosamples [15].
U
AN
Poly(acrylate) SLA, Perfactory1 III GC A: Low cost and customised production. Separation of hexane
Mini SXGA+, L: Column crack-up at higher flow rates, and cyclohexane [16]
M
(EnvisionTEC Inc., dissolution of the printed material in
USA) polar solvents (acetone, alcohols),
blockage of the evaporator and lines by
D
dissolved polymer, high printing failure
TE
rate for large structures.
Poly(acrylate) SLA, Form 2 printer LC A: tailored extraction chamber to Extraction of non-
(Formlabs, USA) accommodate carbon nanofiber/ polymer steroidal anti-
EP
composite membrane. inflammatory drugs
in urine samples
[17].
C
PLA and ABS FDM, RepRap 3D, MS A: Low cost of fabrication, fast analysis Analysis of food,
AC
(Emotiontech, (only 20 s). pharmaceuticals,
France) tequila and mezcal
samples [18, 19]
SLA, MiiCraft ICP-MS A: Low cost (1.50 USD), high permeability, Pre-concentration of
desktop printer rapid column washing and regeneration, Mn, Zn, Ni, Cu, Cd,
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 ACCEPTED MANUSCRIPT

extraction channel (size of (Miicraft, Taiwan) integrated connections. and Pb from

the cuboid 0.4 × 0.4 × 0.2 L: Poor x-y resolution and rough surfaces seawater [20].
mm) of the printed spheres limited the design
to cuboids structures.

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Tube-shaped sorbent for Thermoplastic FDM, ZMorph 2.0.S LC-MS A: Customised tube shape functional Extraction of
SPME (L= 5 mm, O.D. poly(vinyl microfabricator, sorbent glimepiride from

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8.6 mm, wall thickness= 0.2 alcohol) (ZMorph, Poland) water samples [21].
mm) composite

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material (LAY-
FOMM 60)

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Open tube shape sorbent Thermoplastic FDM, ZMorph 2.0.S LC-MS A: Low cost (0.02 USD), customised Steroids extraction

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(wall thickness = 0.2 mm, L poly(vinyl microfabricator, functional sorbent. from human plasma
= 5 mm, diameter= a alcohol) (ZMorph, Poland) [22].
standard microcentrifuge composite

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tube (1.5 mL). material (LAY-
FOMM 60)

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Part of FIA system including Poly(methyl SLA, Form1+, FIA A: Fully automated chemical analysis, Speciation, pre-

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pre-SPE reactor, SPE disk methacrylate) (Formlabs, USA) improved sensitivity (85% increase in concentration and
support, and post column peak height for Fe and ~ 500 fold determination of Fe
reactor with integrating improvement in LOD for chromium VI). [23] and chromium
EP
connectors in water samples
[24].
C

Cylindrical scaffolds (10 x 9 PCL Melt extrusion LC-MS/MS A: Molecularly imprinted polymers (MIPs) Extraction of
AC

mm O.D.) with immobilised bioprinting, with improved retention in the SPE metergoline [25].
polymers as SPE sorbents Bioplotter™ column and reduced back pressure,
(Envisiontec, highly selective muti-analyte extraction
Germany) approach.
Sample holder and tray PLA, ABS FDM, Solidoodle 3rd HPLC-ESI- A: Customised large size sample/solvent Analysis of

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generation and a MS/MS holder (that minimised the sampling time pharmaceuticals and
Fortus 900 mc, to 1 min) and four vial tray, built-in hole metabolites from
(Stratasys, USA) in the sampler allowed switching to whole- body and
surface sampling mode. blade cut rat tissue

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[26].

Minicolumn consisting of Poly(acrylate) SLA, MiiCraft ICP-MS A: No packing required due to Selective extraction

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column holder with fittings BV-007 desktop printer incorporation of nanoparticles (1 % TiO2) of As and Se from
for connectors and inner (Miicraft, Taiwan) in the raw resin prior to printing, highly water and unie

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cuboid stacked packing sensitive analysis. samples [28].

FIA system consisting of a Poly(methyl SLA, Form1+, FIA-UV A: Automated portable system with Determination of Pb

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column (4.5 x 7.8 mm I.D.), methacrylate) (Formlabs, USA) extraction and detection units. in river water

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serpentine mixing coil (500 L: Requires high sample volume for low samples [29].
x 1.5 mm I.D.) and flow cell detection limit requirement.
(optical pathlength = 5 cm)

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Lab on a valve unit Poly(acrylate), Object500 Connex 3 FIA- A: selective quantifications of Cd and Pb Cd and Pb analysis in

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(H×W×D, 66× 100× 17 mm) Acrylic resin printer (Stratasys fluorometer L: Clogging of mixing coil using polyjet water samples [30].
FLGPCL02 Ltd., USA), printer; deformation and microcracking

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Form1+printer from SLA printing resulting in leakage and
(Formlabs, USA), poor optical detection.
EP
RD, L × W × H; RD 1 = 123 × Poly(acrylate), SLA, Viper si2T, (3D HPLC A: Customised RD, easy integration with Monitoring of
C

67 × 42 mm, RD 2 = 100 × Ti-6Al-4V alloy Systems, USA); SLM, separation instrument, thermally stable ketone conversion
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20 × 20 mm, RD 3 = 89 × 27 (Renishaw AM 250 and a polycyclic

× 38 mm system) heterocycle
formation [31]

Reaction vessel (W × L = Polypropylene FDM, 3DTouchTM (3D ESI-MS A: Reusable, customised reactor, Online monitoring of
46.5 × 80 mm, 1.5 mm I.D.) Systems, USA), fabrication of intricate milli- and reaction [34].

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microscale fluidic device with improved

connections and sealing.

Miniaturised reactor (W × L Polypropylene FDM (other details MS A: Embedding of nano-ESI capillary and Online analysis of

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= 15 × 10 mm) not provided) magnetic stirrer bar in the micro-reactor Diels-Alder reaction
during printing. Direct introduction of [35].
sample without external pumps. Fast

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reaction monitoring, minimum ion source
and dead volume. Stability in organic

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solvents and inertness towards various
reagents.

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Flow reactor (Total channel Stainless steel SLM GC-MS, GC- A: Stainless steel allowed the reaction to Difluoromethylation

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length= ∼4 m, I.D= 0.8 mm, FID be carried at extreme temperatures ( ̶ 65 reaction [32].
O.D= 2.4 mm) o
C) and elevated pressures, compatible
with organic solvents, high purity product

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under 2 min.
Cubical reaction chambers Polypropylene FDM, 3DTouchTM (3D 1
H NMR A: Loading of starting material, reagents Chemical synthesis

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(20 mm), connected with 4 Systems, USA) spectroscopy and catalysts during printing with and purification [37]

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mm channels minimum chemical handling, use of
special equipment and low
contamination.
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Reactor-LC interface (size PLA FDM, Ultimaker 2, UPLC-MS A: Better sealing, high pressure tolerance Fast UPLC-MS
not provided) (Ultimaker, (700 psi), low internal volume. analysis of proteins
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Netherlands) [38]
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Platform consisting of two Not provided FDM, (Ultimaker, UPLC or MS A: Reduced analysis time from 3 days to Isolation of glycans
96-well plates for release Netherlands) one day, reduced labour and high from fetuin [39].
derivatisation of throughput due to automation.
carbohydrates

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Separation

Spiral GC columns (450 x 1 Poly(acrylate) SLA, Perfactory GC mobile A: Ordered column geometry, enhanced Ethylene analysis
mm I.D. and 660 x 1 mm Micro HiRes, sensor packing density of the adsorbent, a [40, 41]

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I.D.), dimension: 36 x 36 x (EnvisionTEC Inc., system higher column length/foot print,
18 mm and 40 × 2 × 0.9 USA) integrated connectors, improved
mm, with integrated sensitivity.

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connectors

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Single-piece ABS PJP, Projet HD 3500, LC A: Low cost manufacturing of porous Not provided [44]
chromatography column (3D Systems, USA) beds, internal flow distributers, and [45]
(I.D. = 16mm, wall external fittings. Precise reproducible

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thickness = 2 mm and I.D. = monoliths with controlled structure.

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18 mm, H = 39.3 mm) L: Time consuming support removal
procedure

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Column (600 x 0.9 mm I.D.) Stainless steel, SLM, Realizer SLM HPLC A: Complex spiral geometry of long but Separation of the
housing, external titanium alloy 50, (Realizer GmbH, compact columns phenones [46]
dimension 5 × 30 × 58 mm Germany)

D
L: Rough printed surface, poor separation
efficiency.

Column (600 x 0.9 mm I.D.)
housing, planar footprint 5
× 30 × 30 mm
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Titanium SLM, Realizer SLM HPLC A: Compact, robust column with quick Separation of
50, (Realizer GmbH, temperature equilibration and optimised proteins [47], [48]
EP
Germany) column geometry.
L: Rough printed surface (~ 40µm), low
separation efficiency.
C

TLC stationary phase (50 × Poly(acrylate) SLA, Stratasys A: Quick (25 plates per hour), low cost (1 Separation of a
AC

20 × 1 mm, L × W × H)
TLC stationary phase and
slurry doser (100 × 100 ×
0.2 mm and 95 × 95 × 0.2
Connex Objet 260, euro per plate), low sample and solvent mixture of dyes [49]
(Stratasys Ltd., USA) consumption.
Silica gel as FDM, Prusa i3 TLC A: Quick (30 plates per hour), low cost Analysis of dyes
stationary rework, (0.04 euro per plate). In house fabrication (Oracet Violet 2R,
phase, ABS (Emotiontech, of the customised parts for printer’s Solvent Blue 22,

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mm, L × W × H) and PLA for France) modification. Sudan Red G) [50]

the slurry
doser

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TLC plates (64 × 25 × 0.2 Poly(acrylate), PJP, Objet Eden TLC A: Quick (10 chips per hour), low cost (∼ Separation of dyes,
mm, L × W × H) Veroclear- 260VS, (Stratasys 3.5 USD per plate), customized, reusable proteins (myoglobin,
RGD810 Ltd., USA) plates of varying thicknesses and size. lysozyme) [51].

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Post-separation

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PSI cartridge (L = 50 mm) PLA FDM, Felix, (Felix, MS A: Improved stability to the paper tip, Separation of three
Netherlands) facilitated placement in front of the MS, dyes (Crystal violet,

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integrated features for solvent supply, methylene blue,
disposable. safranin) [52].

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L: Unstable spray generation due to
readjustment of the paper fringes.

M
Photometric detector body PLA FDM, Felix 3.0, (Felix CE, FIA A: Versatile detector for coupling with Analysis of metals in
with integrated slit (slit= 70 robotics, various analytical techniques (HPLC, CE, river water [54]

D
× 400 µm, W × L) Netherlands) FIA).

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Fluorescence detector PLA FDM, CE A: Low cost, precise fabrication of Analysis of
EASY3DMAKER, detector with complex geometries oligosaccharides [55]
(AROJA, s. r. o.,
EP
Czech Republic)
Fluorescence detector with PLA FDM, CE A: Minimises the need for repetitive Analysis of N-
C

LED source of light EASY3DMAKER, analysis in case the signal was too high or glycans,
(AROJA, s. r. o., too low oligosaccharides [56]
AC

Flow cells for
chemiluminescence
detector (body size = 46 ×
34 × 6 mm, L × W × H;
Czech Republic)
Poly(acrylate) PJP, Pro-Jet HD 3000 HPLC A: Enhanced transfer of the emitted light Determination of
3-D Modeler System, from the translucent white printed phenolic amines [57]
(3D Systems, USA) material
L: Difficulties in removal of support

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spiral L = 24 mm) material from the flow channels.

Flow cells for Poly(acrylate) PJP, Eden 260VS, HPLC A: Less tortuous flow, higher H2O2 analysis in urine
chemiluminescence (Stratasys, Australia); chemiluminescence signal, simplified and coffee extract

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detector (spiral cell: 1 mm FDM, Felix 3.0, removal of support material. [58]
I.D., 10 mm O.D., pitch 1.20 (Netherlands) L: Extensive (10 hr) post printing
mm)

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Ion mobility spectrometer PLA, FDM, MakerFarm Stand-alone A: Stand-alone, portable instrument Identification and

SC
polyhydroxyal Prusa i3 design instrument detection of
kanoate, and (Makerfarm, USA) tetraalkylammonium
PETG-CNT bromide salts (TAA),

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benzylamines, and
illicit drugs [59]

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mm-wave detector Stainless steel Details not provided LC A: Low cost, simple, label free detection Detection of sorbitol,
(dimensions not provided) (Materialise, L: Limited dimensions of the device due trans-stilbene and

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Belgium) to printer’s resolution praziquantel [60]

Reaction progress surfaces PLA FDM, MakerBot HPLC A: Communication of complex chemical Visualisation of the

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Replicator (5th concepts HPLC data [63]
generation),

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(MakerBot, USA)
Interface for coupling ABS FDM, UP Plus, 3DP- Microchips A: Easy connection to MS system Monitoring of
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microchips with MS 14-4D, (Beijing glutathione
TierTime oxidation [64]
Technology, Beijing,
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China)
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No. of published papers

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2011 2012 2013 2014 2015 2016 2017 2018
Year

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Fig. 1. The growing number of publications based upon devices and materials produced using 3D
printing technology and applied in one or more modes of separation science.
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Fig. 2. (A, B) CAD drawings of the (A) preconcentrator and (B) a layer of ordered cuboids in the
extraction channel. (C) 3D printed SPE pre-concentrator device. (D, E) 3D printed configuration of
the ordered cuboids (reproduced with permission) [20].
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Fig. 3. Multi-syringe FIA set-up for automated speciation of Fe using a 3D printed two piece device
containing a SPE disk in the middle. V1-V5: Three-way solenoid valves; S1-S4: Syringes; HC: Holding
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coil (reproduced with permission) [23].

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Fig. 4. CAD design, printed model and 20× magnification of SC (a-c) PC (d-f) and HC (g-i) column
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geometries (reproduced with permission) [44].

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Fig. 5. Design of titanium alloy column hardware: (a) 3D printed column hardware, (b) inner view of the design showing the spiral channel, (c)

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chromatogram of a mixture of proteins using the printed column (see the ref for separation conditions and peak identification) (reproduced with
permission) [47].

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Fig. 6. 3D-Printed finger sized PSI cartridge with fast wetting and extended solvent spray properties
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Fig. 7. A) CAD design of the detector and B) 3D printed on capillary photometric detector.
(reproduced with permission) [54].
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Highlights

• Review on impact of 3D printing on the separation sciences

• Impact in areas of sample preparation, handling, chromatographic separations and
detection
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Insight into the future impact and possibilities to transform column technology

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