Macka Talk Pittcon 2016-v2016-03-06-GIVEN-WATERMARKED

Gatherers and Foragers?
Analytical Scientists in the
Quest for Better Light Sources
Pittcon 2016, Atlanta, GA, USA
6-10 March 2016
Professor and Australian Research Council Future Fellow
Australian Centre for Research on Separation Science (ACROSS) and School of Chemistry,
University of Tasmania, Hobart, Australia
http://www.utas.edu.au/chem http://www.across.utas.edu.au
mirek.macka@utas.edu.au
Mirek Macka
Pittcon2016,Atlanta,USA,6March2016

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Pittcon

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What is ’better’ ?
 Depends on the purpose/use  requirements
 Light source properties, size, price, …
Wavelength(s)
Power
 …so you may end up using:
 ‘BIG’: Synchrotron, nuclear reactor, …
 ‘Classical’: incandescent or discharge (W, W-hal., D2, Xe)
Some new ‘classical’ light sources
 Solid state light sources: UV-vis-IR
New areas, strongest growth
 Examples from own research

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Alternative VUV-vis light source?
 IR laser driven light source: LDLS™ (Energetiq)
http://www.energetiq.com/index.php

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Solid state light sources
 The light sources of the 21st century: SSLSs
 SSLSs = LEDs + DLs
 Synergies with new areas of strongest growth
 Mobile technologies
Wearable technologies
 Analysis
 Platforms: Portable / mobile / remote
 Areas:
Biomedical
Environmental
Food & agriculture
 POC
 on-site, in-field

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Solid State Light Sources
 Why SSLSs?
 (Sydney skyline)
“It is expected that optics, also referred to as photonics, will surpass electronics in
the 21st century in terms of the size of the industry reliant on it.”
http://www.sfi.ie/investments-achievements/research-showcase/shedding-light-on-many-subjects/
“In the next decade we will see a massive transformation of the lighting industry
towards energy efficient Solid State Lighting (SSL)”
http://www.photonics21.org/download/olae_sra.pdf

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SSLSs: ‘outside (analytical) chemistry’
 They have many advantages
and still a few weaknesses
 Benefits from large industries
(much larger then chemistry)
 Consumer electronics
 Lighting
 IT & CT
LIFI
 Medical
 Automobile
 Security & military
www.zoneray.com

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Looking ‘out of your box’
 What is this?
iPad / Smartphone sterilisation device!
?  

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Looking ‘out of your box’
 Analytical science is small compared to defence +
‘bread + games’ industries = food, IT, medical...
 Non-analytical usage of LEDs: 3D-agri- and aquaculture,
sterilisation
Vertical Farms, Silicon Chip, March 1012, pp.16-23 siliconchip.com.au

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14
Solid state light sources
 Why use solid state light sources (SSLSs) in science?
 Conventional light sources
Incandescent, discharge lamps as light sources
 SSLSs
Omnipresent, robust, inexpensive, miniaturisation compatible …
1880 2007 

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SSLSs
 LEDs
 Laser diodes (LDs)
 SLEDs
‘hybrid’ properties LED +LD
UV Vis NIR mid-IR
~ 200 nm - 7 µm
Electromagnetic spectrum
schematic representation
Light sources - typical properties:
Traditional: SSLS:
+++ Spectral coverage - + +
(deep-UV to NIR) (from 240 nm up)
+++ Mature well approved technology? - + +
(up to 200 years) (0 to ~40 years)
+ + - Luminosity - + -
- - - Energy conversion, heat production - ++
- - - Radiative heating +++
- - - Miniaturisation compatible? +++
- - - Robustness +++
- - - Life time +++
- - - $$$ +++
- + - Pulsed operation? +++
- + - Noise +++
- + - Future potential +++
0.01nm 1nm 100nm 1µm 1mm 1m 1km

!
 Mirek Macka, Tomasz Piasecki Parmendu K Dasgupta, Light Emitting
Diodes (LEDs) for Analytical Chemistry, Annual Review of Analytical
Chemistry, 7, 183-207, 2014

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16
Dark
Red
Gree
n
Blue
i
"Full colour RGB" LED 621-419
maximum (half-width) nm
0
10
20
30
40
50
60
70
80
90
100
350 400 450 500 550 600 650 700 750
wavelength (nm)
relativeintensity
Blue 438 (70) nm
Green 567 (25) nm
Red 627 (37) nm
SSLSs for analytical devices?
 Advantages of LEDs
 Small, reliable & robust => miniaturised & portable!
 Low-cost: from <$1 to ~$50, but typically ~ $101
 Long life-time: ~105 h & no catastrophic failure
 Very low noise  10-5 AU
Used in various types of optical detectors
(HPLC, FIA etc.)
 Can be operated in a pulsed regime
Can be pulsed
 At extremely fast rates => TRF
 Single-, bi- or tri-coloured LED’s available
 Quasi-monochromatic: w(h/2) ~ 20-70 nm
 ‘Cold light’
5 5.1 5.2 5.3 5.4
Migration time (min)
0.1mAU
a
b


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SSLSs in analysis: history
 Blood oximeter 1972
 Cohen A, Wadswort N
Red/NIR light absorption
Pulsed operation
www.medical-monitors.com
Deoxydated hemoglobin
Oxydated hemoglobin
oximeter.holisticphysio.com

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SSLSs in analysis: history
 Blood oximeter 1972
 Cohen A, Wadswort N
Red/NIR light absorption
Pulsed operation
 1980s: LEDs: IR, RGB
 1990s: explosive growth
 Analytical detection
FIA, LC, CE, chip
1990-1995
 Trojanovicz
 Cardwell, Cattrall & Scollary,
 Huang, Dasgupta, Hauser, Yeung,
Worsfold
www.medical-monitors.comPublications (Web of Science)
0
500
1000
1500
2000
2500
3000
3500
1940 1950 1960 1970 1980 1990 2000 201
Year
No.ofpublications
TS= multidisciplin*
TS= miniaturi*
TS= MEMS
TS= microfluidic*
TI= electrophoresis
TI= liquid* chromatography
TI= gas* chromatography
TI= light emitting diode*
E&CE
HPLC
GC
 Macka M., Andersson P., Haddad P.R., Electrophoresis, 17(12), 1898-1905, 1996
 Mirek Macka, Tomasz Piasecki Parmendu K Dasgupta, Light Emitting Diodes (LEDs) for Analytical Chemistry, Annual
Review of Analytical Chemistry, 7, 183-207, 2014
Deoxydated hemoglobin
Oxydated hemoglobin
oximeter.holisticphysio.com

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‘My’ areas: analytical, separation science
Solid State Light Sources & Miniaturised platforms
SSLSs:
LEDs,
DLs
Platforms:
miniaturised,
portable,
remote
White LEDs:
broad spectrum
light sources
UV LEDs:
portable
fluorescence
microscopy
UV-vis LEDs: photopolymerisations
Chip-CE,
Chip-ITP
Portable
medium-pressure LC
Micro- and small UAV platforms
MEDIA:
Capillaries, chips,
paper(fluidics)
10-6/32 mol/ L
Fluorescein Solution
LEDs:
on-capillary detection
Paperfluidics-inspired
sample preparation

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My ARC FT
SSLSs:
LEDs,
DLs
Platforms:
miniaturised,
portable,
remote
White LEDs:
broad spectrum
light sources
UV LEDs:
portable
fluorescence
microscopy
Chip-CE,
Chip-ITP
Portable
medium-pressure LC
MEDIA:
Capillaries, chips,
paper(fluidics)
10-6/32 mol/ L
LEDs:
sample preparation

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On-capillary photometric detection
 Practical considerations
 In-house designes: simple, robust
Capillary alignment in a Agilent CE optical interface
Vis-LEDs
 Johns, C., et al. Journal of Chromatography A , 927, 237-241, 2001
 Mirek Macka, Tomasz Piasecki Parmendu K Dasgupta, Light Emitting Diodes (LEDs) for Analytical Chemistry, Annual
Review of Analytical Chemistry, 7, 183-207, 2014

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On-capillary detection with LEDs
 UV LED 370 nm
 Buffered chromate electrolyte
 LODs ~10x lower vs. Hg 254 nm
UV LED 370
nm
UV LED 370
nm
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
210 260 310 360 410 460
Wavelength (nm)
AbsorbanceUnits
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
RelativeIntensity
LED
379.5nmHg line
254nm
chromate
absorption
spectrum
0.3
0.8
1.3
1.8
2.3
2.8
3.3
1.95 2.45 2.95 3.45
Migration Time (mins)
Absorbance(mAU)
Cl
-
NO3
-
HCO3
-
(b)
SO4
2- F
-
 King M., Macka M., Paull B., Haddad P. R., Analyst, 127(12), 1564-1567, 2002.

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Photometric
 Deep-UV-LEDs: 255 nm
 Photometric detection
$200-300, ~20-300 mW
EOF
GDP
ADPAMP
0 2 4 6 8 10 12 14
time (min)
absorbance
5mAU
Johns C. et al., Electrophoresis,
2004, 25, 3145–3152
Noise ~0.1 mAU
no optical components
light utilisation <1%
 Stefan Schmid, Mirek Macka, Peter Hauser, UV-absorbance detector for HPLC based on a light-emitting diode,
Analyst, 133, 465-469, 2008 (DOI 10.1039/b715681b)
 Lenka Krcmova, Anna Stjernlof, Sebastien Mehlen, Peter Hauser, Silvija Abele, Brett Paull, Mirek Macka, Analyst,
134, 2394 – 2396, 2009 (DOI:10.1039/ B916081G)

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Photometric
 Deep-UV-LEDs
 Performance
Baseline noise N~0.1mAU
0
5
10
15
20
0 0.1 0.2 0.3 0.4 0.5 0.6
absorbance (AU)
sensitivity(L/mol)
0
500
1000
1500
2000
2500
3000
3500
200 300 400 500 600 700
wavelength (nm)relativeintensity
257.0
 Lenka Krcmova, Anna Stjernlof, Sebastien Mehlen, Peter Hauser, Silvija Abele, Brett Paull, Mirek Macka, Analyst,
134, 2394 – 2396, 2009 DOI:10.1039/ B916081G
Need for better deep-UV-LEDs!
‘suboptimal’ linearity
parasitic vis-band emission

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New generation deep-UV-LED
 High optical output in ‘deep-UV’ @ 255 nm
 Radiometric power 0.57 mW (before ~0.015 mW)
 Negligible parasitic visible range emission
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
200 300 400 500 600 700
Relativeintensity
Wavelength (nm)
255 nm LED
NEW (Optan255H)
OLD (UV TOP 255)
350 450 550 650
Dvorak M. et al., in preparation, 2014

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New generation deep-UV-LED
 High optical output in ‘deep-UV’ @ 255 nm
 Radiometric power 0.57 mW (before ~0.015 mW)
 Negligible parasitic visible range emission
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
200 300 400 500 600 700
Relativeintensity
Wavelength (nm)
255 nm LED
NEW (Optan255H)
OLD (UV TOP 255)
350 450 550 650
7.1 V
~5 V
6.1 V
Dvorak M. et al., in preparation, 2014
0
1000
2000
3000
4000
200 400 600 800
0
1000
2000
3000
4000
200 400 600 800
0
1000
2000
3000
4000
200 400 600 800

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‘Our’ areas: analytical, separation sci.
SSLSs:
LEDs,
DLs
Platforms:
miniaturised,
portable,
remote
White LEDs:
broad spectrum
light sources
UV LEDs:
portable
fluorescence
microscopy
Chip-CE,
Chip-ITP
Portable
medium-pressure LC
MEDIA:
Capillaries, chips,
paper(fluidics)
10-6/32 mol/ L
LEDs:
sample preparation

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Microfluidics has gadgets! 
 Microfluidic platform
 Modular, flexible
Pressure driven flow
Programable multi-channel HV source
Synchronised microscope 1
www.labsmith.com

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Visualization of the separation
 Synchronized video microscope for visualization
available as part of the microfluidic platform

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Detectors: LED (in-house) +
electrochemical (BVT + eDAQ)
Syringe pumps
(LabSmith) Switchable valves
(LabSmith)
Injector (VALCO)
4-20 nL
Capillary column: monolith
(C18 - Merck Chromolith)
1
2
4
3
Mobile phase A
Switching valves
A1
A2
B2
B1
B
A
Mobile phase B
Waste
Sample
Injector
Data
acquisition
Capillary column
Pressure sensor
LED detector
~$700
~$1,000
~$700
Design
~$200
ca 25 cm
 Li Y., Dvorak M, Nesterenko P., Stanley R., Nuchtachvorn N., Kujovska Krcmova L, Aufarova J., Macka M, Anal. Chim. Acta, 896,
166-176, 2015

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Performance of the LC system
 Isocratic Separations (ion-pair RP LC)
Column: 300 x 0.1 mm ID
Eluent: 50 mM ammonium acetate -
acetonitrile 50/50 (v/v)
F=0.5 µL/min.
Detection: LED on-capillary photometric
detector (254 nm)
Sample: 60 µM methyl 4-hydroxybenzoate
(MHP), 0.16 mM ethyl 4
hydroxybenzoate (EHB), 0.16 mM
propyl 4-hydroxybenzoate (PHB),
and 0.17 mM butyl 4
hydroxybenzoate (BHB)
RSD < 1% (peak area), efficiency N ~ 80,000 TP
 Li Y., Dvorak M, Nesterenko P., Stanley R., Nuchtachvorn N., Kujovska Krcmova L, Aufarova J., Macka M, Miniaturised medium
pressure capillary liquid chromatography system with flexible open platform design using off-the-shelf microfluidic components,
Anal. Chim. Acta, 896(8), 166-176, 2015

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 Gradient Separations: MeOH
Eluent: A=50 mM NH4AC (pH 5),
B=methanol, linear gradient 0 to
100% B in 8.3 min,
F=1 µL/min.
Detection:
355 nm LED on-capillary photometric detector
Sample: Dichromate 5 mM, tartrazine 1 mM,
orange G 1 mM, naphthol yellow 1 mM
and brilliant yellow 1 mM, 20 nL
Compounds Tr
(min)
RSD (n=8)
Tr
Peak area
(mAU*t)
RSD (n=8)
Peak area
K2Cr2O7 1.19 1% 77 3%
Tartrazine 2.86 5% 45 5%
Orange G 3.45 4% 29 6%
Naphthol Yellow 3.76 3% 23 5%
Brilliant Yellow 6.17 1% 79 4%
-10
-5
0
5
10
15
20
25
0 2 4 6 8 10 12
mAu
Time (min)
Tartrazine
Cr2O7
2-
Orange G
Naphthol Yellow
Brilliant Yellow
100 %

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 Gradient Separations: MeCN
-10
-5
0
5
10
15
20
25
0 2 4 6 8
mAu
Time (min)
Coumarin
60%
100%
Cr2O7
2-
Tartrazine
Naphthol Yellow
Brilliant Yellow
Sudan III Sudan IV
Martius Yellow
Eluent: A=50 mM NH4AC (pH 5),
B=acetonitrile, linear gradient 0 to 60%
B for 5 min, then 100% B for 3 min.
F=1 µL/min.
Detection:
355 nm LED on-capillary photometric detector
Sample: dichromate 4 mM, tartrazine 1.5 mM,
naphthol yellow 1.5 mM, brilliant yellow
0.5 mM, martius yellow 2 mM, coumarin 1
mM, sudan III 0.5 mM and sudan IV 0.5
mM, 20 nL
 Li Y., Dvorak M, Nesterenko P., Stanley R., Nuchtachvorn N., Kujovska Krcmova L, Aufarova J., Macka M, Miniaturised medium
pressure capillary liquid chromatography system with flexible open platform design using off-the-shelf microfluidic components,
Anal. Chim. Acta, 896(8), 166-176, 2015

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LEDz-cell
Ball lens
Ball lens
Silicon photodiode
Optics?
LED-z-cell
 LED-z-cell photometric detector
 Considerations
• Li Y. et al., unpublished results

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LED-z-cell
 LED-z-cell photometric detector
Eluent
Slit
Tubular LED holder
defining distance
LED
Silica photodiode

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-0.1
0.1
0.3
0.5
0.7
0.9
1.1
0 2 4 6 8 10 12
Absorbance(mAU)
Time (min)
LED detector
Commercial detector
45 nL
LED-z-cell
 Comparison of separation performance (254 nm)
 The gradient separation of genetic amines: L-Dopa 9 ppm, L-tyrosine 18 ppm, norfenefrine 14 ppm,
phenylephrine 18 ppm and tyramine 18 ppm, linear gradient 5 mM to 35 mM methanesulfonic acid
(MSG) 0 to 12 min, flow rate 12 µL min-1. Column: CS19, 20 cm x 200 µm i.d., injection volume: 400 nL.
-0.1
0.1
0.3
0.5
0.7
0.9
1.1
0 2 4 6 8 10 12
Absorbance(mAU)
Time (min)
LED detector
Commercial detector
180 nL

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On-capillary detection with LEDs
 Quality of detection optical setup easily checked
 Effective pathlength
 Stray light %
Instrument Detector linearity Effective
upper limit (AU) pathlength (mm)
Agilent 3DCE 1.2 64.6
AB 270A-HT 0.75 60.5
Waters CIA 0.175 49.70
2
4
6
8
10
12
14
16
18
20
1 10 100 1000 10000
absorbance (mAU)
Sensitivity[AU·L·mol-1]
Agilent 3D-CE
Applied Biosystems 270A-HT
Waters CIA
 Johns C., Macka M., Haddad P.R., King M., Paull B., J. Chromatogr. A, 927(1-2), 237-241, 2001
 Johns C., Macka M., Haddad P.R., LC-GC Europe, 16(5), 290, 292, 294-295, 2003

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Photometry and photometric detection
 Quality of detection optical setup easily checked:
Sensitivity vs. absorbance graph
 Effective pathlength
 Stray light %
Linearity evaluation
0
500
1000
1500
2000
2500
0 0.1 0.2 0.3 0.4 0.5 0.6
Chromate [mol/L]
Absorbance[mAU]
Agilent Technologies 3D CE
Applied Biosystems
Waters CIA
0
2
4
6
8
10
12
14
16
18
20
0.1 1 10 100 1000
Chromate [mmol/L]
Agilent Technologies 3D CE
Applied Biosystems
Waters CIA
0
2
4
6
8
10
12
14
16
18
20
1 10 100 1000 10000
Absorbance [mAU]
Agilent Technologies 3D-CE
Applied Biosystems
Waters CIA
 Johns C., Macka M., Haddad P.R., King M., Paull B., J. Chromatogr. A, 927(1-2), 237-241, 2001
 Johns C., Macka M., Haddad P.R., LC-GC Europe, 16(5), 290, 292, 294-295, 2003

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BeckmanInstruments
AgilentTecnologies
deltaDOT
PrinceTechnologies
Sebia
MicroTech*
Lumex
UnimicroTechnologies*
CEResources**
0
10
20
30
40
50
60
Brand
Weight[kg]
0
0.05
0.1
0.15
0.2
0.25
0.3
Volume[m3
]
W [kg] V [m3]
 Portable platforms - still rare species…
 Example: CE
 Markéta Ryvolová, Jan Preisler, Dermot Brabazon, Mirek Macka, Portable capillary-based (non-chip) capillary
electrophoresis: current state, instrumentation and future development, TRAC, 29(4), 339-353, 2010
Analytical instrumentation: Portability?

Portable platforms - still rare species…
 How portable is commercial instrumentation?
 Example: CE
 Markéta Ryvolová, Jan Preisler, Dermot Brabazon, Mirek Macka, Portable capillary-based (non-chip) capillary
electrophoresis: current state, instrumentation and future development, TRAC, 29(4), 339-353, 2010

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SSLSs:
LEDs,
DLs
Platforms:
miniaturised,
portable,
remote
White LEDs:
broad spectrum
light sources
UV LEDs:
portable
fluorescence
microscopy
Chip-CE,
Chip-ITP
Portable
medium-pressure LC
MEDIA:
Capillaries, chips,
paper(fluidics)
10-6/32 mol/ L
LEDs:

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Research chips in a commercial chip-CE
 Agilent Bioanalyzer 2010
 DNA-chip vs. in-house-designed chips
DNA-chip In-house-chips
Chipcaddie

- 43 -
 Chips: Complex challenges
 Optical detection: Fluorescence primarily

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 Details to be found in these publications:
1. Petr Smejkal, Ákos Szekrényes, Markéta Ryvolová, Frantisek Foret, András Guttman, Fritz Bek, Mirek Macka, Chip-based CE for
rapid separation of 8-aminopyrene-1,3,6-trisulfonic acid (APTS) derivatized glycans, Electrophoresis, 31(22), 3783-3786, 2010,
http://dx.doi.org/10.1002/elps.201000457
2. Nantana Nuchtavorn, Fritz Bek, Mirek Macka, Worapot Suntornsuk, Leena Suntornsuk, Rapid separations of Nile blue stained
microorganisms as cationic charged species by chip-CE with LIF, Electrophoresis, 33 (9-10), 1421-1426, 2012,
3. Nantana Nuchtavorn, Petr Smejkal, Michael C Breadmore, Philip Doble, Fritz Bek, Rosanne M Guijt, Frantisek Foret, Leena
Suntornsuk, Mirek Macka, Exploring chip-CE-LIF platform flexibility: Separations of fluorescent dyes by chip-based non aqueous
CE, J. Chromatogr.A, 1286, 216–221, 2013, http://dx.doi.org/10.1016/j.chroma.2013.02.060
4. Petr Smejkal, Michael C. Breadmore, Rosanne M. Guijt, Frantisek Foret, Fritz Bek, Mirek Macka, Isotachophoresis on a chip with
indirect fluorescence detection as a field deployable system for analysis of carboxylic acids, Electrophoresis, 33(21), 3166-3172,
2012, http://dx.doi.org/10.1002/elps.201200141.R1
5. Petr Smejkal, Michael C. Breadmore, Rosanne M. Guijt, Jakub Grym, Frantisek Foret, Fritz Bek, Mirek Macka, Separation of
carboxylic acids in human serum by isotachophoresis using a commercial field-deployable analytical platform combined with in-
house glass microfluidic chips, Anal. Chim. Acta, 755, 115– 120, 2012, http://dx.doi.org/10.1016/j.aca.2012.10.022
6. Petr Smejkal, Michael C. Breadmore, Rosanne M. Guijt, Frantisek Foret, Fritz Bek, Mirek Macka, Analytical isotachophoresis of
lactate in human serum using dry film photoresist microfluidic chips compatible with a commercially available field-deployable
instrument platform, Anal. Chim. Acta, 803, 135–142, 2013, http://dx.doi.org/10.1016/j.aca.2013.01.046
7. Petr Smejkal, Danny Botteus, Michael C Breadmore, Rosanne M Guijt, Cornelius F Ivory, Frantisek Foret, Mirek Macka,
Chip-ITP: a review, Electrophoresis, Special Issue Bioanalysis, 34(11), 1493-1509, 2013, http://dx.doi.org/10.1002/elps.201300021

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Detection: Concurrent multi-detection
 LED photometric on-capillary
 250-660 nm, baseline noise ~0.0001 AU
 Macka M., Andersson P., Haddad P.R., Linearity evaluation in absorbance detection: The use of light emitting diodes for on-
capillary detection in capillary electrophoresis, Electrophoresis, 17(12), 1898-1905, 1996
 Johns C., Macka M., Paul R. Haddad, Design and performance of a light-emitting diode detector compatible with a
commercial capillary electrophoresis instrument, Electrophoresis, 25(18-19), 3145-3152, 2004,
 Lenka Krcmova, Anna Stjernlof, Sebastien Mehlen, Peter Hauser, Silvija Abele, Brett Paull, Mirek Macka, Deep-UV LEDs in
photometric detection: A 255 nm LED on-capillary detector in capillary electrophoresis, Analyst, 134, 2394 – 2396, 2009,
http://dx.doi.org/10.1039/B916081G
 Marketa Ryvolová, Jan Preisler, Pavel Krásenský, František Foret, Peter C. Hauser, Brett Paul, Mirek Macka, Single Point of
Detection Combined Contactless Conductometric, Photometric and Fluorimetric on-Capillary Detector for Capillary
Separation Methods, Anal.Chem., 82(1), 129-135, 2010, http://dx.doi.org/10.1021/ac902376v
C4D electrodes
capillary
Second fluorescence
excitation (400 mm OF)
Fluorescence emission
pick-up (300 mm OF)
Optical fiber (OF)
SMA adapter
PD detector
(optical fiber)

 Triple detection
C4D+PD+FD
 Identical light source
for PD and FD: 470 nm
 Noise~<0.1mAU
1,24
1,25
1,26
1,27
1,28
1,29
1,3
0 2 4 6 8 10 12
Migration Time (min)
Signal(AU)
PD
tartrazine
3,5
4
4,5
5
5,5
0 2 4 6 8 10 12
Signal(V)
C4D
CO3
2-
MES
His
EOF
K+
tartrazine
28
33
38
43
48
53
0 2 4 6 8 10 12
Signal(mV)
fluorescein FD
3-in-1 combined C4D + PD + FD
CONDITIONS:
Sample: standard mix
Capillary: 75 mm, 31.5/39 cm
BGE: 20 mM CHES buffer pH 9
Voltage : 12 kV, Injection: 7 cm,10 s
C4D: 100 kHz
PD: LED 470nm, no filter @ 30 mA (2.7 mW),
300 mm /50mm in/out fibres
FL: Ex.: LED 470nm, no filter @ 20 mA (2.7 mW),
300 mm /300 mm excit./pick-up fibre
 Markéta Ryvolová, Jan Preisler, Pavel Krásenský, František Foret, Peter C. Hauser, Brett Paul, Mirek
Macka, Anal.Chem., 82(1), 129-135, 2010, DOI 10.1021/ac902376v

 Design
 1 point of detection
 Fibre optics
 SMA adapter
C4D electrodes
capillary
Second fluorescence
excitation (400 mm OF)
Fluorescence emission
pick-up (300 mm OF)
Optical fiber (OF)
SMA adapter
PD detector
(optical fiber)
C4D
electrodes
Capillary
SMA adapter
(PD input & pick-up, FD
input)
Fluorescence pick-
up (optical fiber)
Detector assembly platformDetector assembly cover
A B
3-in-1 combined C4D + PD + FD
 Markéta Ryvolová, Jan Preisler, Pavel Krásenský, František Foret, Peter C. Hauser, Brett Paul, Mirek
Macka, Anal.Chem., 82(1), 129-135, 2010, DOI 10.1021/ac902376v

- 48 -
Rapid design using 3D-printing
 CAD  3D-printing  characterisation
 Extrusion (Felix)

- 49 -
LED-IF: on-capillary detection - comparison
 3-in-1
High Brightness LED 460 nm
@ 700 mA, 3.1 lm, optical fibre 1 mm
~27 mW
LOD = 4 × 10-8 M LOD = 1 × 10-9 M
Ultra bright 3mm LED 470 nm
@ 30 mA, 3.6 V, 5000 mcd, 35º
~ 2.7 mW
LEDs vs. lasers
 LODs 10-10 M vs. 10-12 M
 Prices ~ € 10 vs. ~ € 5,000

- 50 -
LEDIF: µLEDs
 LEDs - the smallest light sources ever
 Micro LEDs with diameter ~15 μm
 LED chip – 1 mm2
 LED Cluster – 450 μm diameter
 170 µLEDs/ 400 μm optical fibre
 Integrated filters, micropackaged
15 mm
 Tanriseven, S., Maaskant, P., Corbett, B., Applied Physics Letters, 92, 123501, 2008
 Vaculovičová, M., Akther, M., Maaskant, P., Brabazon, D., Macka, M., Fibre coupled micro-light emitting diode array
light source with integrated band-pass filter for fluorescence detection in miniaturised analytical systems,
Anal.Chim.Acta, 871, 85-92, 2015

- 51 -
Back metalFilter
n -
contact
Light generating
active area
Dielectric
p - contact
Semiconductor (GaN)
Bond metal
Optical fibre
SiO2/HfO
LEDIF: µLEDs
 Microfabrication & micropackaging
 Custom designed integrated interfrence filters
400 450 500 550
0.0
0.5
1.0
S16 (without filter)
S19 (with filter)
Normalisedcounts
Wavelength, nm
Normalised emission spectra
 Tanriseven, S., Maaskant, P., Corbett, B., Applied Physics Letters, 92, 123501, 2008
 Vaculovičová, M., Akther, M., Maaskant, P., Brabazon, D., Macka, M., Fibre coupled micro-light emitting diode array
light source with integrated band-pass filter for fluorescence detection in miniaturised analytical systems,
Anal.Chim.Acta, 871, 85-92, 2015

- 52 -
LEDIF: µLEDs
 LOD (fluorescein) = 5x10-9 mol/L @ ~50 µW
Outlook: SLEDs!
35
40
45
50
55
60
200 400 600 800 1000 1200 1400
Migration Time [s]
IF[a.u.]
Sample: Maltooligosaccharide ledder
(Diluted by H2O 1:10)
Capillary: 75 mm, 30/40 cm
BGE: 50 mM acetate pH 4.7 + 20% DNA gel
Voltage : -10 kV, Injection: 7 cm, 15 s
300 mm pick-up fibre
µLED Device: S 19 (with filter) @ 20 mA
capillary
mLED fibre mLED chip
pick-up fibre
to PMT
 Vaculovičová, M., Akther, M., Maaskant, P., Brabazon, D., Macka, M., Fibre coupled micro-light emitting diode
array light source with integrated band-pass filter for fluorescence detection in miniaturised analytical
systems, Anal.Chim.Acta, 871, 85-92, 2015

- 53 -
SSLSs:
LEDs,
DLs
Platforms:
miniaturised,
portable,
remote
White LEDs:
broad spectrum
light sources
UV LEDs:
portable
fluorescence
microscopy
Chip-CE,
Chip-ITP
Portable
medium-pressure LC
MEDIA:
Capillaries, chips,
paper(fluidics)
10-6/32 mol/ L
LEDs:
sample preparation

- 54 -
LEDs as a broad-spectrum source?
 UV-vis-NIR LEDs as a multi-chip
‘all-wavelengths-in-one’ LEDs
 $?
 White LEDs?
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
200 300 400 500 600 700 800 900 1000 1100
LineIntensity
Wavelength, nm
Normalized Emission Spectrum of UV-Vis-NIR LEDs
355 nm
390 nm
415 nm
458 nm
490 nm
540 nm
570 nm
595 nm
625 nm
660 nm
690 nm
720 nm
750 nm
780 nm
820 nm
850 nm
880 nm
920 nm
980 nm
Wavelength [nm]
200 300 400 500 600
LampCounts
0
2000
4000
6000
8000
10000
12000
14000
16000
18000
Deutrium Lamp
Luxeon LED

- 55 -
LED as a broad-spectrum light source
 White LED as a broad-spectrum light source ?
 White phosphorus LED
Mounted on a
D2-lamp base

Tomasz Piasecki
 Piasecki T., Breadmore M.C., Macka M., White LEDs as broad spectrum light sources for spectrophotometry:
Demonstration in the visible spectrum range in a diode-array spectrophotometric detector,
Electrophoresis, 31(22), 3737-3744, 2010 (DOI 10.1002/elps. 201000341)

- 56 -
 White LED as a broad-spectrum light source for a DAD
- photometric detection for CE
 Luxeon LED powered to currents of 0-400 mA.
0 mA
50 mA
100 mA
200 mA
400 mA

- 57 -
 Spectra
 Luxeon (colour-coded)
 Deuterium lamp (black)
Wavelength [nm]
200 300 400 500 600
LampCounts
0
2000
4000
6000
8000
10000
12000
14000
16000
18000
Deutrium Lamp
Luxeon LED
current: 50-400 mA
~5nm (thermal effect)
Time [min]
0 2 4 6 8
Absorbance[mAU]
0
2
4
6
8
10
Deuterium Lamp
Time (min)
0 2 4 6 8
Absorbance(mAU)
0
2
4
6
8
10
Luxeon LED

- 58 -
White LED as a broad-spectrum source
 Photometric detection in CE – D2 Lamp
 Photometric detection in CE - LEDs
 Better spectra quality for the LED lamp! 
 Only vis-range available 
Wavelength [nm]
200 300 400 500 600
Absorbance(mAU)
0
2
4
6
8
FITC
Wavelength [nm]
200 300 400 500 600
0
2
4
6
F
Absorbance(mAU)
Wavelength [nm]
200 300 400 500 600
0
4
8
12
16
20
24
PR
Absorbance(mAU)
200 300 400 500 600
0
2
4
6
8
OG
Wavelength [nm]
Absorbance(mAU)
Wavelength (nm)
200 300 400 500 600
Absorbance(mAU)
0
2
4
6
8
FITC
Wavelength (nm)
200 300 40
0
500 600
0
2
4
6
8
F
Absorbance(mAU)
Wavelength (nm)
200 300 400 500 600
0
4
8
12
16
20
PR
Absorbance(mAU)
Wavelength (nm)
200 300 400 500 600
0
2
4
6
OG
Absorbance(mAU)
 Piasecki T., Breadmore M.C., Macka M., White LEDs as broad spectrum light sources for spectrophotometry:
Demonstration in the visible spectrum range in a diode-array spectrophotometric detector,
Electrophoresis, 31(22), 3737-3744, 2010 (DOI 10.1002/elps. 201000341)

- 59 -
SSLSs:
LEDs,
DLs
Platforms:
miniaturised,
portable,
remote
White LEDs:
broad spectrum
light sources
UV LEDs:
portable
fluorescence
microscopy
Chip-CE,
Chip-ITP
Portable
medium-pressure LC
MEDIA:
Capillaries, chips,
paper(fluidics)
10-6/32 mol/ L
LEDs:
sample preparation

- 60 -
LED-Imaging
 Microscopy
 Portable
 Low-cost
 USB Microscopes
 Dino-Lite http://www.bigc.com/
 Cheap (~$50) Chinese

- 61 -
LED-Imaging
 Microscopy
 Portable
 Low-cost

- 62 -
LED-Imaging
 Microscopy
 Portable
 Low-cost

- 63 -
SSLSs: ‘non-analytical’
 LEDs as light sources in many areas
 Plant science
Wavelength-selective light
 Biotechnology
Green algae
 Photochemistry
 Photolithography – chip microfabrication
e. g. Breadmore MC et al.

- 64 -
64
Where use SSLSs?
 LEDs, LDs, SLEDs
 Optical detection
Fluorimetric
Photometric
 Single-colour
 White
 Photoinitiated
polymerisations
of monoliths
 Microphotochemistry
White LEDs:
broad spectrum
light sources
Single-colour or white LEDs:
photopolymerised monolith
LED-IF and diode LIF
Single-colour LEDs:
quasi-monochromatic
light sources
Single-colour LEDs:
photochemistry

- 65 -
Where use SSLSs?
 LEDs, LDs, SLEDs
 Optical detection
Fluorimetric
Photometric
 Single-colour
 White
 Microphotochemistry
 Photoinitiated polymerisations
of monoliths
White LEDs:
broad spectrum
light sources
Single-colour or white LEDs:
photopolymerised monolith
LED-IF and diode LIF
Single-colour LEDs:
quasi-monochromatic
light sources
Single-colour LEDs:
photochemistry

- 66 -
Photopolymerisations
 LEDs as light sources
 UV: 255 nm: Silvija Abele, Fu-Qiang Nie, František Foret, Brett Paull, Mirek
Macka, UV-LED photopolymerised monoliths, Analyst, 133, 864 - 866,
2008, DOI:10.1039/B802693A
 Vis: 660 nm: Zarah Walsh, Silvija Abele, Brian Lawless, Dominik Heger,
Petr Klán, Michael C. Breadmore, Brett Paull, Mirek Macka, Photo-initiated
Polymerisation of Monolithic Stationary Phases Using Visible Region LEDs,
Chem. Commun., (48), 6504 – 6506, 2008, DOI:10.1039/B816958F
 Vis: 470 nm: Zarah Walsh, Pavel A. Levkin, Brett Paull, Frantisek Svec and
Mirek Macka, Visible light initiated polymerisation of styrenic monolithic
stationary phases using 470 nm light emitting diodes, J.Sep.Sci., 33(1),
61-66, 2010, DOI:10.1002/jssc.200900624
 UV: 365 nm: Silvija Abele, Smejkal Petr, Yavorska Oksana, Frantisek Foret,
Mirek Macka, Evanescent wave photoinitiated polymerization of open-
tubular capillary monolithic columns, Analyst, 135 (3), 477-481, 2010,
DOI:10.1039/b920789a
Monolith reviews:
 Knob, R., Kulsing, C., Boysen, R.I., Macka, M., Hearn,
M.T.W., Surface-area expansion with monolithic open
tubular columns, TrAC, 67, 16-25, 2015
 Zarah Walsh, Brett Paull, Mirek Macka, Inorganic
Monoliths in Separation Science: A Review, Anal. Chim.
Acta, 750, 28-47, 2012

- 67 -
2008, DOI:10.1039/B802693A
61-66, 2010, DOI:10.1002/jssc.200900624
DOI:10.1039/b920789a
Monolith reviews:
Acta, 750, 28-47, 2012

- 68 -
 Abele S., Nie F.-Q., Foret F., Paull B., Macka M., Analyst, 132, 864 - 866, 2008
Monoliths: Synthesis with UV LEDs
 Microfluidic chips – easily made in situ where needed
 http://www.rsc.org/publishing/journals/AN/article.asp?doi=b802693a

- 69 -
2008, DOI:10.1039/B802693A
61-66, 2010, DOI:10.1002/jssc.200900624
DOI:10.1039/b920789a
Monolith reviews:
Acta, 750, 28-47, 2012

- 70 -
Red-LED photoinitiation in polyimide..
 “Photoinitiated polymerisation of monolithic stationary
phases in polyimide coated capillaries using visible
region LEDs”
 Zarah Walsh, Silvija Abele, Brian Lawless, Dominik Heger, Petr Klán, Michael C. Breadmore, Brett Paull, Mirek Macka,
Photo-initiated Polymerisation of Monolithic Stationary Phases Using Visible Region LEDs,
Chem. Commun., (48), 6504 – 6506, 2008, DOI: 10.1039/B816958F

- 71 -
 Modelling/simulating light penetration
 Counter-intuitive patterns
10% 1% 0.1%
85% 75% 65%
 Piasecki, T., Macka, M., Paull, B., Brabazon, D, Numerical model for light propagation and light intensity distribution
inside coated fused silica capillaries, Optics and Lasers in Engineering, 49 (7), 924-931, 2011

- 72 -
2008, DOI:10.1039/B802693A
61-66, 2010, DOI:10.1002/jssc.200900624
DOI:10.1039/b920789a
Monolith reviews:
Acta, 750, 28-47, 2012

- 73 -
 Evanescent wave
photoinitiation (EWP)
 EWP in transparent
PTFE-coated
fused silica
capillaries
Conditions
• initiator - DAP
• LED - 365 nm
• polymerisation time 15 min
Results
5 cm of capillary filled with polymer, but
top end is whiter, bottom end – paler
(polymer only around the wall proved by SEM)
Spectra not matched
Evanescent wave photoinitiation

- 74 -
 EWP
365 nm
LED
20 mm 0 mm4.5
mm
8.5
mm
11.5
mm
15.5
mm
Evanescent wave photoinitiation
 Silvija Abele, Smejkal Petr, Yavorska Oksana, Frantisek Foret, Mirek Macka, Evanescent wave photoinitiated
polymerization of open-tubular capillary monolithic columns, Analyst, 135 (3), 477-481, 2010, DOI:10.1039/b920789a

- 75 -
LEDs in photochemistry
 LEDs in the photoswitching of photochromic molecules
MC-Co Complex Absorbance Data
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
300 400 500 600 700 800
Wavelength (nm)
Absorbance
0.00005M
0.0001M
0.0005M
0.001M
0.005M
0.01M
0.05M
0.1M
MC - no Co present
370nm
525nm
 Walsh, Z., Scarmagnani, S., Benito-López, F., Abele, S., Nie, F.-Q., Slater, C., Byrne, R., Diamond, D., Paull, B., Macka, M.,
Photochromic spiropyran monolithic polymers: Molecular photo-controllable electroosmotic pumps for micro-fluidic
devices, Sensors and Actuators, B: Chemical, 148 (2), 569-576, 2010
 Scarmagnani, S., Walsh, Z., Slater, C., Alhashimy, N., Paull, B., Macka, M., Diamond, D., Polystyrene bead-based system
for optical sensing using spiropyran photoswitches, J.Mater.Chem., 18 (42), 5063-5071, 2008

- 76 -
Future?
Every
perspective
is personal
and
relative…

- 77 -
LEDs: Deep-UV?
 From ~0.01 mW  to 1-3 mW 
HICSUNTLEONES
0.01
0.1
1
10
100
1000
200 300 400 500 600 700
Wavelenght [nm]
Radiometricpower[mW]
€0.00
€100.00
€200.00
€300.00
€400.00
€500.00
€600.00
€700.00
€800.00
Price[€]
Single chip LED power emitters
Multichip LED high power emitters
Single chip LED price
Multichip LED price
 Mirek Macka, Tomasz Piasecki Parmendu K Dasgupta, Light Emitting Diodes (LEDs) for Analytical Chemistry,
Annual Review of Analytical Chemistry, 7, 183-207, 2014

- 78 -
Future of SSLSs in science
 SSLSs: Deep-UV?
 The “Alloy Road” to deep-UV-LEDs: AlN

- 79 -
Future of SSLSs in science
 LEDs: Spectra and prices now and into the future
Emission spectra for LEDs and LDs
0
0.5
1
200 300 400 500 600 700 800
Wavelength (nm)
RelativeIntensity
256 nm
281 nm
370nm
401 nm
473 nm
523 nm
Laser 532 nm
634 nm
660 nm
$
101
102
100
$
W

- 80 -
SSLSs: New ‘cool gadgets’…
 High power low cost lasers
 http://www.wickedlasers.com/
 UV-LED torches
 395 nm
 http://au.element14.com/night-searcher/nsnuvled395/torch-led-uv/dp/1823957?Ntt=1823957&CMP=i-55c5-00001402
 365 nm, 395 nm
5W (electrical input)
 http://www.farnell.com/datasheets/606066.pdf

- 81 -
SSLSs: New ‘cool gadgets’…
 Applications: photosynthesis – plant growth
 Spectrum needed:
 Chlorophyl spectrum
http://en.wikipedia.org/wiki/Chlorophyl
 Red + blue

- 82 -
Future?
 Light sources  SSLSs
 LEDs
 Deep-UV (<300 nm)
Higher power! (W) + ‘clean’ spectra
VUV ? (<180 nm) ???
 MIR (2-7 um)
 LDs
 Parallel the success of LEDs
‘Specialties’ e.g. QCLs (IR)
 New areas, strongest growth
 Vacuum UV (VUV), deep-UV, IR light sources
 Portability
 Imaging – all wavelengths

- 83 -
Acknowledgements
 SPECIAL THANKS TO: 
$ Australian Research Council (ARC):
Future Fellowship Level 3 (Professorial)
$ Australian Endeavour Fellowship
$ University of Tasmania, Hobart, Australia,
ACROSS & School of Physical Sciences
$ Agilent, Waldbronn, Germany
University Relations Grant
$ Royal Jubilee Scholarship,
Mahidol University, Thailand
$ Grant Agency of the Czech Republic
OTHER COLLABORATORS :
Companies: LabSmith, USA; Knauer AG, Germany
Academic: Prof Leena Suntornsuk, Mahidol University, Bangkok, Thailand
…….
AU$
€
CZK
THB
AU$
AU$

- 84 -
UTAS: PhD scholarship available
 Tasmania: Beautiful & mild climate
 Follow Charles Darwin 
 Numerical modelling
Thank you!

- 85 -
Thank you!
 The presentation available at
https://www.researchgate.net/profile/Mirek_Macka

Macka Talk Pittcon 2016-v2016-03-06-GIVEN-WATERMARKED

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