User Guide

For an introduction to the application and use of Certified Reference Materials in absorption spectrophotometry



These Reference Materials can be used to qualify the Stray Light (or Stray Radiant Energy) of ultraviolet spectrometers according to the Filter Ratio or Meilenz method. This method is described in ASTM E 387 and is recommended for instrumentqualification by the US Pharmacopeiain its Chapter <857>, published in May 2015. Chapter <857> is mandatory from 1st May 2016. The European Pharmacopoeia and most other regulatory bodies recommend the Specific Wavelength method.


Description and Discussion

A range of cut-off filter solutions that allow stray light to be checked at a range of wavelengths from 200 nm to 390 nm. Each liquid filter is permanently sealed by heat fusion into a 10 mm high quality far UV quartz cell and a 5mm cell of thesame liquid is used as a back-off in the instrument’s reference beam. A spacer is supplied so that the 5mm cell can be held securely in the reference beam cell holder. Starna alkali halidestray light Certified Reference Materials are prepared in accordance with ASTM E-387.

Stray light, also called Stray Radiant Energy or Power, is anylight reaching the detector that is outside the Spectral BandWidth selected for analysis by the monochromator. It can be due to optical imperfections or stray reflections withinthe monochromator itself or to light leaks or other effects in the rest of the optical system. As the detector cannotdiscriminate between the analytical wavelength and thestray light, the stray light contributes to the detector signal and introduces an error in the measured absorption. The stray light is not absorbed even at high concentrations of the absorbing species, so its effect is a negative deviationfrom the linear relationship between concentration and absorbance (the Beer-Lambert law) on which most quantitative determinations are based.

Stray light is wavelength and instrument dependant. It can be present at any wavelength but is most noticeable when the energy throughput of the system at the analytical wavelength is relatively low, for example in the far UV region, and any stray light will be comparatively more significant. At these wavelengths, any deterioration in the instrument optics or UV light source will exaggerate the apparent stray light, so It is desirable to check it even if the instrument is not to be used in the far UV, as it is an excellent way of monitoring the condition of the instrument optics.

The usual way of assessing stray light is to measure, at the desired analytical wavelength, a sample that totally absorbs the radiation at that wavelength, but transmits at all other wavelengths. Any light detected by the instrument is then stray light. In the traditional method, known as the “Specified Wavelength” method, the cut-off filter appropriate to the analytical wavelength is scanned against a water blank.

This method was also accepted by the US Pharmacopoeia until May 2015, when ca new chapter <857> was issued,recommending the Solution Filter Ratio method, also
known as the Meilenz method. In this method the reference materials are measured not against water, but against a 5 mm path length cell containing the same solution. This has theeffect of “backing off” the measured absorbance, resulting in a peak. Spectra obtained with a Potassium Chloride reference material using the two methods are shown below:

The differential absorbance value (ΔA) at the peak is related empirically to the Stray-Light level (s) by

s = 0.25 x 10 –2*ΔA

Practically, this means that an instrument exhibiting 1% stray light would give a differential absorbance value at the peak of > 0.7A. This is the instrument qualification requirement of the USP.

Typical spectra


These typical spectra were recorded on three instruments with different stray light characteristics:

The peak wavelength is different on each instrument due to their different stray light characteristics, but in all three cases the absorbance maximum is greater than 0.7A, so all these instruments satisfy the USP stray light requirement.

For comparison purposes, the equivalence of the two test methods is indicated in the table:

An instrument giving an absorbance of >2.0 A using theSpecified Wavelength method should give a differentialabsorbance of >0.7A using the Filter Ratio method.

The working ranges of the solutions as cited in the USP are:

Filter Ratio Absorbance (Δ A) Specified Wavelength Absorbance (A)
0.3 1.3
0.5 1.6
0.7 2.0
1.0 2.6
1.5 3.6
2.0 4.6
2.5 5.6

Material  CONCENTRATION  Usable range Catalogue Number 

Sodium Nitrite, 10mm and 5 mm cells

5% aqueous

300 - 385 nm  RM-SN/5

Acetone, 10mm and 5 mm cells

Spectroscopy grade

250 - 320 nm  RM-AC/5

Potassium Iodide, 10mm and 5 mm cells

1% aqueous

210 - 259 nm  RM-KI/5

Sodium Iodide, 10mm and 5 mm cells

1% aqueous

210 - 259 nm  RM-SI/5

Potassium Chloride, 10mm & 5 mm cells

1.2% aqueous

190 - 205 nm  RM-KC/5 



Catalogue Number


Material  Catalogue Number 

Universal Stray Light reference set, EP and USP compliant


Material  Catalogue Number 

Sodium Nitrite, 10mm and 5 mm cells


Acetone, 10mm and 5 mm cells


Potassium Iodide, 10mm and 5 mm cells


Sodium Iodide, 10mm and 5 mm cells


Potassium Chloride, 10mm & 5 mm cells