Starna scientific

Quality Assurance in the Analytical Laboratory ‘Setting the Standard’

Spectrofluorometer Wavelength Validation

& Performance Check
Starna 6BF Series Polymer Block References

Purpose
These materials may be used on a routine
basis to calibrate, compare and monitor
the performance of Fluorescence
Spectrophotometers. p-Terphenyl, Ovalene,
Tetraphenylbutadiene and Anthracene in
PMMA are cited in USP chapter <853> for
wavelength validation. .

Description and Discussion

Six polymer blocks containing seven fluorescent compounds whose
spectra cover a broad spectral range with excitation range from 260
to 600nm and emission range from 370 to 630 nm. This wide spectral
range enables the user to select a reference material with broadly
similar spectral properties to those of the analyte, ensuring a reasonable
spectral overlap and measurable signals without changing important
parameters such as slit widths and wavelength settings.

REF COMPOUND APPROXIMATE PEAK EXCITATION EXCITATION PEAK EMISSION EMISSION RANGE
NO MOLAR WAVELENGTH (NM) RANGE (NM) WAVELENGTH (NM) (NM)
CONCENTRATION
1 Anthracene 1 x 10-5 360 260 - 320 402 370 - 470
Napthalene 6 x 10-5 290 290 - 390 330 310 - 400
2 Ovalene 2 x 10 -7
342 300 - 460 482 450 - 570
3 p-Terphenyl 5 x 10 -7
295 270 - 350 338 310 - 410
4 Tetraphenylbutadiene 3 x 10-7 348 290 - 390 422 370 - 540
5 Compound 610 1 x 10 -6
440 360 - 490 475 420 - 570
6 Rhodamine 2 x 10 -7
562 500 - 600 573 530 - 630

The solid polymethyl methacrylate (PMMA) matrix provides a stable environment for the fluorescent compounds. Being solids, there can be no
evaporation of solvents, providing long term stability.
 Spectrofluorometer Performance Validation
Polymer Block References
Ex Ex
Em Em
Ex Ex
Em Em
Typical excitation and emission spectra are shown below.

nm 310 320 330 340 350 360 370 380 390 400 410 420 430 440 450 460 470 nm 270 280 290 300 310 320 330 340 350 360 370 380 390 400

Starna Naphthalene Reference

nm 310 320 330 340 350 360 370 380 390 400 410 420 430 440 450 460 470 nm Starna
270 280 290 Anthracene
300 310 320 330 Reference
340 350 360 370 380 390 400
Intensity

Intensity
Ex
Ex Em
Em
Ex Ex Ex
Ex Em
Em Em Em
Ex
Ex Em Ex
Em Em

nm 272 285 297 310 322 335 347 360 372 385 397 410 nm 310 320 330 340 350 360 370 380 390 400 410 420 430 440 450 460 470 nm 270 280

Wavelength nm Wavelength nm
nm 285 300 315 330 345 360 375 390 405 420 435 450 465 480 495 510 525 540 555 570
nm 272 285 297 310 322 335 347 360 372 385 397 410
nm 270 280 290 300 310 320 330 340 350 360 370 380 390 400
nm 285 300 315 330 345 360 375 390 405 420 435 450 465 480 495 510 525 540 555 570

Starna Ovalene Reference

nm 310 320 330 340 350 360 370 380 390 400 410 420 430 440 450 460 470 nm Starna p-Terphenyl
270 280 290 Reference
300 310 320 330 340 350 360 370 380 390 400

Ex
Ex
Em
Em
Ex
Intensity

Intensity

Ex Ex Em Ex Ex
Em Em Ex EmEmEx
Em Ex Ex
Em
Ex
Em Em
Em
nm 272 285 297 310 322 335 347 360 372 385 397 410 Ex
Em
nm 315 330 345 360 375 390 405 420 435 450 465 480 495 510 525 540 555 570 585 600 nm 285 300 315 330

nm 315 330 345 360 375 390 405 420 435 450 465 480 495 510 525 540 555 570 585 600 nm 370 380 390 400 410 420 430 440 450 460 470 480 490 500 510 520 530 540
nm 310 320 330 340 350 360 370 380 390 400 410 420 430 440 450 460 470 nm 270 280 290 300 310 320 330 340 350 360 370 380 390 400
nmnm310
285320
300330
315 330
340 345
350360
360375
370390 405
380 420400
390 435410
Wavelength 450nm
465 480
420 430 495
440510
450525
460540 555 570
470 nm 270 280 290 300 Wavelength
310 320 330 nm340 350 360 370 380 390 400
nm 272 285 297 310 322 335 347 360 372 385 397 410 nm 370 380 390 400 410 420 430 440 450 460 470 480 490 500 510 520 530 540

nm 285 300 315 330 345 360 375 390 405 420 435 450 465 480 495 510 525 540 555 570

Starna Compound 610 Reference Starna Tetraphenylbutadiene Reference

Ex
Em
Ex
Em
Ex
Em Ex Ex
Em Ex
Em
EmExEx Ex
Intensity

Intensity

EmEm Em Ex
Em
nm 315 330 345 360 375 390 405 420 435 450 465 480 495 510 525 540 555 570 585 600 Ex
Em

nm 510 520 530 540 550 560 570 580 590 600 610 620 nm 370 380 390 4
nm 272 285 297 310 322 335 347 360 372 385 397 410
0
nm 272 285 297 310 322 335 347 360 372 385 397 410
nm 510 520 530 540 550 560 570 580 590 600 610 620 nm 285 300 315 330 345 360 375 390 405 420 435 450 465 480 495 510 525 540 555 570
nm nm
370315
380 390
330 345400
360410
375 420 430 420
390 405 440435
450450
460 470
465 480480
495490
510 500 510 555
525 540 520570
530585
540
600 nm 285 300 315 330 345 360 375 390 405 420 435 450 465 480 495 510 525 540 555 570

Wavelength nm
Wavelength nm
nm 370 380 390 400 410 420 430 440 450 460 470 480 490 500 510 520 530 540

Starna Rohodamine B Reference

Ex
Em

Ex
Ex Em Ex
Em Em
Ex
Ex Em Ex
Intensity

Em Em

nm 510 520 530 540 550 560 570 580 590 600 610 620

nm 315 330 345 360 375 390 405 420 435 450 465 480 495 510 525 540 555 570 585 600
nm 315 330 345 360 375 390 405 420 435 450 465 480 495 510 525 540 555 570 585 600
nm 370 380 390 400 410 420 430 440 450 460 470 480 490 500 510 520 530 540

nm 510 520 530 540 550 560 570 580 590 600 610 620 nm 370 380 390 400 410 420 430 440 450 460 470 480 490 500 510 520 530 540

Wavelength nm

Ex
Em Ex
Em
Spectrofluorometer Performance Validation
Polymer Block References

Certification and Documentation Accreditation

STARNA warrants that all its Reference Materials have been
produced in accordance with ISO17034. The ISO 17034
compliant certificate supplied states that the Reference
Material has the required stability and homogeneity
characteristics and is fit for its intended use
Starna Scientific is accredited to both ISO Guide 34 (4001)
Starna Scientific is accredited to both ISO 17034 as a
Reference Material producer, and ISO/IEC 17025 as a
Calibration Laboratory for optical reference measurements.
Starna Scientific’s manufacturing facility is accredited to the
ISO 9001 Quality Management System with BSI.

How to Order
COMPOUND CATALOGUE NUMBER
Set of 6 PMMA Fluorescence References 6BF
Napthalene/Anthracene 6BF-STD/1
Ovalene 6BF-STD/2
p-Terphenyl 6BF-STD/3
Tetraphenylbutadiene 6BF-STD/4
Compound 610 6BF-STD/5
Rhodamine B 6BF-STD/6

Note: these catalogue numbers relate to polymer blocks in standard rectangular cell format. For other
formats please contact Starna at sales@starna.com

‘Setting the Standard’

www.starna.com
sales@starna.com
+ 44 (0) 20 8501 5550