Lecture delivered during CSI Symposium-2016

1. Analytical Instrumentation – From
Concept to Commercialization
Milton L. Lee
Department of Chemistry and
Biochemistry

2. Analytical Chemistry
March 1981

4.  Novel – not previously disclosed
 Not obvious to someone trained in the art

5. Patent application
February 1982

6. Lee Scientific
Incorporated
May 1985

7. Artist’s Drawing of
SFC Pump

9. Utah
Technology
Finance
Corporation
August 1985
$50,000

10. Analytical Chemistry
September 1985

11. Dionex invests
$0.5 million
March 1986

14. Patent challenged
February 1987

16. SFC Technology Transfer Timeline
• 1981 First report published
• 1982 Patent application filed
• 1984 Patent approved
• 1985 Lee Scientific incorporated
• 1985 Received $50,000 from UTFC
• 1985 Received financing from Dionex
• 1986 First instrument shipped
• 1987 Patent re-validated by patent office
• 1988 Lee Scientific sold to Dionex

17. The Rest of the Story
• Dionex discontinued SFC,
but continued with SFE
• SFE morphed into
accelerated solvent
extraction (ASE)
• Lee Scientific SFC
continues with Selerity
Technologies
• SFC morphed into the
Selerity elevated
temperature LC product
line
Series 4000 SFC
Polaratherm

18. Analytical Chemistry
December 1991

20. Laboratory Prototype TOFMS

21. Patent application
August 1990
Sensar
Corporation
1990

24. Iulia Lazar
1993-1997

30. TOFMS Technology Transfer Timeline
• 1990 Patent application filed
• 1990 Sensar incorporated
• 1991 First report published
• 1991 Patent issued
• 1992 First instrument sold
• 1993 R&D 100 Award received
• 1999 Company sold to LECO

31. Portable Instrumentation
Weight/Size
Power
Consumables/waste
Performance in
“harsh environments”

32. Portable GC-MS System
• Rapid results
• Low analysis cost
• Light-weight/small
• Battery-operated
• Minimal sample handling
• Minimal consumables/waste
• Easy operation
• Operable in variety of
environments
• Sampling/sample introduction
straightforward

33. Lammert et al., Int. J. Mass Spectrom. 2001, 212, 25
Toroidal Ion Trap MS
Miniature trap
ro = 2 mm
Trapping V = ~ 1 kVp-p
Conventional size trap
ro = 1 cm
Trapping V = ~ 16 kVp-p

34. Electrodes and Analyzer Assembly
Entrance/Exit Slits = 0.011”

35. S.A. Lammert et al.,
J. Am. Soc. Mass
Spectrom. 17, 916
(2006)

36. CB007 Solid Model

37. GC-TMS System
Batteries
Diaphragm
Pump
Turbo
Molecular
Pump
GC System
He Gas
Cartridge
ElectronicsVacuum Chamber
with Toroidal Ion Trap

38. LTM Capillary GC
Typical GC Conditions:
• Column: MXT-5 (5 m x 0.1
mm x 0.4 mm)
• Mobile Phase: Helium
• Head Pressure: 25 psig
• Column Max Flow: ~0.4
scc/min @ 20oC
• Split-splitless Injection:
Selectable split ratios: 0, 10,
50, and 60 to 1
• Program: 50oC (hold 10 s) to
300oC (hold 10 s) @
120oC/min
• GC-TMS run time: <5 min
(including GC cooling cycle
and data processing)
GC column
Injection port
Cooling fan Electronics

39. Resistively-heated Column Bundle
Column bundle cross section
5 m stainless steel capillary
0.1 mm i.d.
0.4 micron MTX-5

41. 2008 R&D 100 Award
The
“Oscars” of
Invention

42. TRIDION™-9
15”
15.5”
9”
LTM gas chromatograph
5 m x 0.1 mm x 0.4 mm MTX-5
Fast temperature programming
50 - 300ºC @ 120ºC/min
Toroidal ion trap
Electron ionization
Electronic pressure control
45 to 500 m/z @ 10 scans/s
Low power
~120 W peak
~ 60 W average

43. 171 175
173
Performance
• Resolution:
< 1 amu FWHM across
mass range
• Scan speed:
~10 scans/s
~1 s
Bromoform

44. Standard Mixtures Using PDMS
Reservoir
Glass wool
PDMS
Valve cap
Vial
Absorbed calibration
compounds

45.  Reliable GC RT and MS calibration
 Solvent-less standards for on-site validation
 No dilution or liquid standard preparation necessary
 No solvent peaks or solvent related interference
 Compatible with SPME for solvent-less injections
 Closed system: standards absorbed into a granular solid
matrix
 Long-term stable and reproducible
Chromatogram of the Calion PV Mix
Acetone
Methylenechloride
Methyl-tert-butylether
Heptane
Methylcyclohexane
Toluene-d8
Perchloroethylene
Bromopentafluorobenzene
Bromoform
1,2-Dibromotetrafluorobenzene
Methylsalicylate
Tetrabromoethane
Tetradecane
Reproducibility and stability data for a
single Calion vial
Calion Calibration Mixture

46. Comp
No.
After 50 After 100 After 200
1 98.30 96.61 93.21
3 96.16 92.33 84.65
5 96.05 92.10 84.20
7 98.02 96.03 92.06
9 99.62 99.24 98.47
11 99.93 99.86 99.71
13 100.00 100.00 100.00
Calculated Percentages of Calibrants
Remaining in Vial After N Samples Taken
30 s, 25oC, DVB/PDMS extraction

47. Law-Enforcement Applications
Leary, P. E.; Dobson, G. S.; Reffner, J. A. Appl Spectrosc 2016, 70, 888
Home-made explosives
Triacetone triperoxide (TATP)
Arson fire accelerants
Gasoline
Counterfeit drugs
Volatile organics in Viagra
tablets
Direct SPME
Headspace SPME

48. Rounded tip
with hole
Loose-fitting
angle-cut wire
Packing 1
Packing 2
19-gauge
needle
Side Hole
Loose-fitting wire
Teflon sleeve
Spring
Tight-fitting wire
Sealant
Needle Trap
Sampling/Sample Preparation
 Pawliszyn, et al.
 Torion/PerkinElmer

49. History
2001 Torion Technologies founded
2003 Funding received from DTRA
2007 Guardion-7 introduced
2008 Torion receives R&D 100 Award
2011 Tridion-9 introduced
2015 Torion acquired by PerkinElmer

50. Why Miniaturize LC?
 Point-of-care/portability
 Laboratory bench space
 Solvent use and waste
 Inlet to MS

51. Compact LC Development Criteria
 Hand portable
 Line voltage and battery operated
 Low flow rate (1-1000 nL/min)
 Ultra-high pressure (up to 16,000 psi)
 Low extra-column volume
 Sensitive UV-absorbance detector
 Easily interfaced to MS
 Simple construction
 High performance
 Easy to operate
Tranxend Corporation, 2016

52. Manual
remote
control
24 V DC
power supply
MicroLynx (I-CPM)
stepper motor
controller
RS-232 interface
Remote
switch box
24 V DC
power supply
E2CA
valve
controller
Motor box
Piston
Seals
360 µm fittings
Stator
Rotor
Drive shaft
Pump cavity
Nano-flow Pumping System

53. y = 1.0006x - 0.0002
R2 = 0.9995
0
0.02
0.04
0.06
0.08
0.1
0.12
0 0.02 0.04 0.06 0.08 0.1 0.12
Measuredflowrate(µL/s)
Theoretical flow rate (µL/s)
Calibration using Eksigent flow calibration
capillary
• 16 readings at each flow rate for
pure H2O & ACN/H2O (70:30 v/v)
• Flow rate error < 0.061%
• Acceptable error = 1%
Flow-rate Reproducibility

54. Deep-UV LED Detector
 Small and compact
 No need for expensive optical
lenses
 Quasi-monochromatic
 High stability – No need for
reference cell
 Long life
 Low power requirement
S. Sharma et al., Anal. Chem., 2015, 87,1381
LED holder
LED with ball lens (6.4 mm dia.)
260 nm band-pass filter
Ball lens (3 mm dia.)
Slit (razor blade pair)
Photodiode
Capillary (0.150 mm i.d.)
Thorlabs

55. Detector Linearity and LODs
Analytes
Concentration
range
R2 Sensitivity
LOD
(ppb)
Sodium
anthraquinone-2-
sulfonate
24.6 nM – 50.4 µM >0.99 0.99 7.6
Adenosine-5-
monophosphate
87.9 nM – 22.5 µM >0.99 1.01 31
DL-tryptophan
299 nM – 0.61
mM
>0.99 0.99 60
LOD improvement factors:
Anthraquinone 200
Adenosine 230
Tryptophan 60

56. Gradient UHPLC Nano-flow Pumping System
Valve A Valve B
Pump A
Pump B
Valve C
Injection
syringe
Solvent A
Solvent B
Waste
Electronic box
 Non-split flow
 15,000 psi pressure
 5.6 kg weight
 24 V battery
 30 µL pump volume
(each)
 30+ nL stop-flow injector

57. Separation of Pharmaceuticals
0 2 4 6 8 10 12 14 16
0
50
100
150
200
250UVabsorbance(mV)
Retention time (min)
1
2
3
4
1. Acetaminophen
2. Aspirin
3. Ibuprofen
4. Indomethacin
15 cm x 150 µm i.d. PEGDA monolith
A = acetonitrile (pH = 2.5)
B = water (pH = 2.5)
A-B gradient from 10-100% B (5 min)

58. Integrated Column and ESI Tip
• 10-12 cm long, 75 µm i.d.
• Waters BEH 1.7 µm particles
(Mike Fogwill, Waters)
• Small after-column volume
• Tip end opening ~15 µm
Packed with
home-pulled tip
Packed with New
Objective tip

59. Coupling of Nanoflow LC to Orbitrap MS
 3 kV spray voltage
 25 µm i.d. transfer line
 ~100-200 nL dwell volume
 300-500 nL/min

60. Analysis of BSA Tryptic Digest
25%
5%
40%
 30 min linear gradient
from 5-30% ACN
 12 cm x 75 µm i.d. column
 500 nL/min
 1.0 µL injection
 25 min compensation time
 40 min separation window
 ~20 s base peak width
 Large peak capacity
 MSMS database identified over 90% peptides

61. What’s Next?
“The Road goes ever on
and on
Down from the door where it
began.
Now far ahead the Road
has gone,
And I must follow, if I can,
Pursuing it with eager feet,
Until it joins some larger
way
Where many paths and
errands meet.
And whither then? I cannot
say.”
The Hobbit

62. Seems like there is always another adventure!

63. September
2011
“Cornucopia” of
Students and
Colleagues

64. Students from India
Pankaj
Aggarwal
2014
Sonika
Sharma
2015
Abhijit
Ghosh
2015+