Making Measurements using

Fiber Optics and the Cary 60

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MARK FISHER APPLICATIONS ENGINEER

AGILENT TECHNOLOGIES

21 May 2015

Applications for which Fiber Optics might be Ideal

Working Solution

• In Situ Measurements (kinetics)

• Working with small volumes

• Can not bring Sample to Instrument (Art Restoration)

• Desire to reduce transfer of Samples to Cuvettes

• Measurement Needs to be made a distance from Instrument

• Measurements need to be made in an inert atmosphere

(glove box)

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Cary 100

UV-Vis NIR Spectroscopy Solutions

The Agilent Cary Family

Cary 300

Cary 4000

8453

Cary 5000/6000i

Cary 60

Cary

Eclipse

Perf

orm

ance

Routine to Research

“A word from our sponsors”

What are Fiber Optics

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Fiber Optics

Cladding

Central Core

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How Does Optical Spectroscopy Work From The

Instrument View

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Source Energy

Selection Sample Detector Display

Why the Cary 60 ?

Cary 60 is uniquely designed :

• Small beam spot size

• Room Light Immunity

• Short Pulse Duration (key for photosensitive materials)

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Regular Cuvette Holder for Cary 60

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Beam Size as It Enters Sample Compartment

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Beam Size at Focus in Middle of Cell Holder

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Fiber Optic Couplers and Immersion Probes

Fiber Optic Couplers for the Cary 60

Immersion Fiber Probe Design

Immersion Fiber Probe Replaceable Tips (reusable)

Stainless Steel with replacement tips

Torlon Probe with replaceable tips

Quartz Probe (does not come with replaceable tips)

Versa Probe with Disposable Tips (NOT reusable)

Micro Probe (does not come with replaceable tips)

Fiber Optic Couplers for Cary 60

The Fiber Optic Dip Probe is a

trouble-free alternative to a

conventional sipper.

The Fiber Optic Coupler allows

you to connect a fiber optic

probe to suit your application.

Immersion Fiber Probe Design

• Stainless Steel Probe

• Torlon Probe

• Quartz Probe

• Micro Probe

• Versa Probe

Immersion Fiber Probe Replacement Tips

(reusable)

Replaceable Tips come in a

various selection of path lengths

and are cleanable and reusable.

These tips can be used on the

Stainless Steel and Torlon

Immersion Probes.

Cary 60 Differentiators

Fiber Optics No more need for expensive cuvettes!

Improve workflow by taking the instrument to the sample

Measure cold, hot and odd-shaped samples

Measure

cold

samples

directly from

the

refrigerator!

Improve

workflow

and

minimize

sample

preparation

APPLICATION NOTES AVAILABLE:

5990-7863EN Measuring purity of low volumes of DNA at 4 ºC with fiber optics microprobe

5990-7945EN Determination of drug tablet concentration in pharmaceutical applications for drug development

5990-7864EN Simple, automated measurements of the photocatalytic properties of colorimetric species

Application Focus

No compromise in data quality,

strong linear correlation:

relationship between

absorbance and concentration

Abs

DNA concentration (ng/µl)

y = 0.0198x R² = 0.9996

0.0

0.5

1.0

1.5

2.0

0 20 40 60 80 100

DNA scans using fiber

optic microprobe

www.agilent.com (Publication Number 5990-7863EN)

Short Pathlength Solution using Fiber Optics

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• Easy to clean and load

• No sample carry over

• Recovery of precious samples

• Different path length options

• Can be thermostated

• Ease of use:

• Pipette sample

• Place lid over sample

• Take measurement

• Wipe clean

Tray Cell Performance Benefits

Measure High Abs With Short Paths

Tray Cell Accuracy Cary 4000 vs Cary 60

Hellma TrayCell – linearity

20

Excellent linear

performance to 2.3abs

Detect down to 0.001

mg/mL of BSA (using 10mm

pathlength cuvette)

Dedicated software

Hellma TrayCell

10mm cuvette 1.0mm cap 0.2mm cap

max abs 3.3 abs 2.3 abs 2.3 abs

dilution factor 10x 50x

equiv abs 23 abs 115 abs

No dilution necessary 0 to 115 abs!

More Options for Measurements of Microvolumes

by UV-Vis

For customers who need to measure the concentration or purity of small volumes

of precious samples such as nucleic acids or proteins - there is a better

alternative! With a design focused on ease of use, and superior performance in

sensitivity and reliability, the Agilent solution is the Cary 60 and the ultra-

microvolume cuvette accessory, which now has more options available for a

wider range of sample volumes and concentrations.

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Cary 60 With Microscope

Measurements made in a Inert Atmosphere

(glovebox)

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Cary 60 with Fiber Optic Coupler

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Fiber Optics to the Glovebox Feed-through

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Glovebox Feed-Through

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Inside Glovebox

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Cuvette Holder Inside Glovebox

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Sample Measurement Outside Glovebox

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Diffuse Reflectance Measurements using Fiber

Optics

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Cary 50 With Remote DRA and Camera

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Useful in Art Restoration Projects

• Allows measurement to be made at painting

• Mount Accessory on an X-Y stage

• Able to fix distance from art work to accessory

• Use Color software to determine L*a*b or other color values

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Fiberoptic Dissolution

Options with the Cary 60

Fiber Optic Benefits vs. Multicell Systems

• Very fast sampling times,

30s+

• More time points ideal for

early formulation work and

immediate release

formulations

• Able to analyze samples

where filtration is not

possible/practical (<0.45um)

• Can work in extremely low

volumes

• No Consumables

Why Fiber Optics on the Cary 60?

• Highest optical transmission of any system on the market allowing it to easily read in high turbidity solutions

• Can lose over 90% transmission and still have more transmission than other fiber optic systems on the market

• Flashlamp lamp preserves life of fiber optic probes

Agilent’s New Technology – Fiber Optic Probes.

• Specially designed for dissolution testing.

• 600 micron silica/silica Fibers for optimized transmission down to 190 nm.

• 1, 2, 5, 10, 20mm tips available.

• Streamlined to minimize hydrodynamic influence.

Fiber Optics not just for UV-Vis Measurements

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Cary Eclipse Fluorescence Spectrometer

The Power of Xenon…

• Unique Xe flashlamp technology

• Measure small volume samples

• Fiber optics

• Room light immunity: unique,

Varian patented technology

• Eliminates photo-degradation

• Long lamp lifetime

Application focus

• Biochemical applications

• Academia

• Industrial chemistry

Special Thanks

Special Thanks to Dr. Jillian L. Dempsey with the Department of

Chemistry at the University of North Carolina and her graduate

student, Robin Knauf, for providing access to their Cary 60 and

glove box as well as a sample they are interested in for use in

data collection for this presentation

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Thanks For Your Patience And Good Humor

Shortly You Will Receive A Link

To Down Load The Presentation

My Contact is Mark.Fisher@Agilent.com

Thanks Again