Good DERE Measurement Practice

Customer Seminar
The Philippines
September 2012
Agenda

 What is Density and How is it Measured?

 Good Density Measurement Practice
 What is Refractive Index and How is it Measured?
 Good Refractive Index Measurement Practice

2
Density

Definition

m Mass m

Density
rho
r =
V Volume V

The density is mass divided by volume

3
Density

Temperature
1.0050

1.0000

0.9950
Density [g/cm³]

0.9900

0.9850

0.9800

0.9750

0.9700

0.9650

0.9600
0 10 20 30 40 50 60 70 80 90
Temperature [°C]

Example: Influence of temperature on the density of water

The density depends on the temperature

4
Density

Units
Density m
Mass r =
 d(20) = = g/cm3
Volume V

Specific Gravity
d20 Sample g/cm3
 d(20/20)*) = = = no dimension
d20 Water g/cm3

d20 Sample g/cm3

 d(20/4)*) = = = no dimension
d4 Water g/cm3

The density is expressed as g/cm3 or without dimension as

specific gravity
5
Density

Absolute and apparent density

Absolute density
 Determined in vacuum or with digital density meter

Apparent density
 Determined in air (with pycnometer) without correcting for buoyancy effect
 Many old tables are still based on apparent density!
 Some dig. meters can display results in dA, but result may still differ slightly
Examples
Apparent density True density
Air (20 ºC) 0.00000 g/cm3 0.00120 g/cm3
Water (20 ºC) 0.99715 g/cm3 0.99821 g/cm3

Absolute and apparent density is not the same (make sure

terms are clear when comparing results)
6
Density

How to measure density?

Density meter

Density kit
Pycnometer

Hydrometer

METTLER TOLEDO

Old New

7
Density

Hydrometer

 Dip hydrometer into sample

 Read density off the scale
 Calibrated to show absolute density

8
Density

Hydrometer

+ Easy method
+ Cheap
+ Quick measurement
- Thermostat required
- Large sample volume
- Manual reading
- Operator dependent result
- Breakable glassware
- Accuracy questionable
- Limit of error: around 0.001, but only if….

9
Density

Pycnometer

 Tare balance with empty glass container

 Fill glass container with sample to known
volume
 Weigh glass container with sample and
calculate result
100 cm3
 Usually glass container contains air when
balance is zeroed  apparent density is
measured

120 g
Density = = 1.2 g/cm3
100 cm3

10
Density

Pycnometer

+ Cheap
- Requires much experience
- Density must be calculated
- Thermostat required
- Large sample volume
- Time consuming
- Operator dependent result
- Breakable glassware
- Accuracy questionable
- Limit of error: around 0.001 (more theoretically, but only if….)

11
Density

Density kit

 Weigh a glass body of a defined

volume in air
 Add liquid (sample)
 Weigh the glass body in the sample
and calculate result (absolute density)
 For solids: weigh sample in air and
then in water (or other liquid of know
density)  difference depends on
density of sample (buoyancy)

M in air – M in liquid
Density =
Volume body

12
Density

Density kit

+ Automatic reading
+ Printout of result (GLP)
+ Just an accessory to a balance
+ Possibility to determine density of solids
- Large sample volume
- Not possible to thermostat
- Accuracy: not better than 0.001 g/cm3

13
Density

Digital density meter

 Fill U-tube with sample

 Let U-tube oscillate with different frequencies till
resonance is observed
 Resonance frequency of U-tube is inversely
proportional to density of sample inside U-tube
 Oscillation value at resonance (T-value) is
inversely proportional to resonance frequency
and direct proportional to density

14
Density

Digital density meter

Thermo conductive block Peltier element
(heat & cool)

Oscillator oscillating U-tube

temperature sensor
15
Density

Digital density meter

+ Quick measurement
+ High accuracy achievable *)
+ Small sample volume
+ Built-in thermostat (no water bath)
+ Temperature and stability control
+ GLP (print out of results)
- Expensive (initial investment only)

Inject the sample

*) Press start
Attention with viscous samples!
Read result
(or fully automatic)

16
Density

Viscosity error

 Viscous samples attenuate the oscillation of the U-tube

 Result: The resonance frequency is reduced (oscillation value at
resonance is increased) – the digital density meter shows a density
that is too high

1 Orange juice
Viscosity error

1
2 2 Olive oil
3 Detergent
4 Honey

3 4

sample viscosity
17
Density

Viscosity error

 To correctly measure samples with a viscosity > 50 mPa*s (like

olive oil) with an accuracy of 0.0001 g/cm3 this viscosity error
must be corrected for
 Such a viscosity error can be around 0.001 g/cm3 at the max.
 Some density meters are equipped with an automatic viscosity
correction feature (depending on measured density, correct
error compensation is automatically applied)
 3 decimal place density meters do not require such a viscosity
correction (accuracy not high enough anyway)

18
Density

Summary

Sampling Sample Operator Temperature Initial

time volume dependency control investment

19
Agenda

 What is Density and How is it Measured?

 Good Density Measurement Practice
 What is Refractive Index and How is it Measured?
 Good Refractive Index Measurement Practice

20
Good Density Measurement Practice

Overview

 Instrument Stability
 Sample Preparation
 Sampling
 Cleaning
 Measuring
 Tests
 Adjustment

21
Instrument Stability

Temperature control via Peltier element

 Example DM40 (4 place instrument)

Upper part

19.979
Temperature / °C

19.978

19.977

19.976

19.975
11:45:36 11:48:29 11:51:22 11:54:14 11:57:07 12:00:00 12:02:53 12:05:46 12:08:38 12:11:31

Time / hh:mm:ss

 Highly accurate and stable – deviations of 2 to 4 mK only!

 Don‘t switch off – keep instrument running

22
Instrument Stability

Temperature control via Peltier element

 Designed to ensure and maintain absolutely stable and correct

temperature
 Not made for quick changes
 At power up or change of temperature, wait 15 to 30 minutes to
reach first roughly stable level
 If possible, avoid temperature changes
- Reduces waiting time
- Increases accuracy and repeatibility

23
Sample Preparation

Common error sources

 Evaporation issues and gas containing samples

 Viscous and solid samples at room temperature

24
Sample Preparation

Evaporation issues and gas containing samples

 Do not heat up samples over boiling point

- Volatile fraction can be lost (different density)
 Degas samples not containing volatile fractions
- Gas bubbles (e.g. air) form in samples naturally
 Heat samples in closed vials
 Controlled heating

25
Sample Preparation

Viscous and solid samples at room temperature

 Samples need to be heated and kept at desired temperature

 Filling samples must be possible without introducing bubbles
 Use heating control system (e.g. SC1H)

26
Sampling

Common error sources

 Cell is not dry

 Filling is not correct
 Cross-contaminations are introduced

27
Sampling

Cell is not dry or contaminated

 Make sure cell is cleaned and completely dry

 If contaminations in the cell (sample, solvent)
- Next result is wrong
- Especially bad if next measurement is an
adjustment with air
 Make sure to use the correct rinsing liquids
 Even very small quantities of rinsing liquids Remaining Caused
or residues of previously measured samples rinsing liquid in measuring error in
in the cell can cause substantial measuring uL g/cm3
errors 10 0.001500
 Example: If cell of density meter was rinsed 1 0.000150
with ethanol and not completely dried, and
then measurement of water is performed, 0.1 0.000015
error due to remaining rinsing liquid is as 0.01 0.000001
follows

28
Sampling

Cell is contaminated

 Do not just barely fill the cell

- Contaminations remain in the cell
 Control the filling amount
 Ensure usage of correct rinsing liquids

29
Sampling

Cell is contaminated

 Fill cell at a low speed (5 – 10 cm per second)

 Make sure that sample comes out of cell for at least 15 cm
 Ensure that contamination is pushed out at the front
 Only new sample left in cell

30
Cleaning

Common error sources

 Wrongly selected cleaning solution

 Wrong or uncomplete drying

31
Cleaning

Select correct cleaning solution for 1st rinse

 Which solvent to select for first cleaning (rinse 1)?

- “How is the glassware cleaned?”
 When samples are water, or water based: use water
 When samples are acids (concentrated): use LOTS of
water
 When they also contain fats, or oily components
(salad oil) or when samples are alkaline: use 0.3 to
0.5 % Deconex solution
- Deconex dissolves in water, acetone, and ethanol!
 For petrochemical samples: use Toluene, or petrol
ether mixtures

32
Cleaning

Select correct cleaning solution for 2nd rinse

 Which solvent to select for second cleaning (rinse 2)?

- Must dissolve the first cleaning liquid
 If water or Deconex solution was used as first rinse
- Use acetone
- If acetone is not allowed: use ethanol
- For ethanol: must be 100% (not 96% and not denaturated)
 If toluene or similar was used as first rinse
- At room temperature: use low boiling petrol ether mixture
or acetone
- At higher temperatures (>30 deg. C.): use hexane or
similar

33
Cleaning

Drying duration around room temperature

How to determine the right drying time?

 Run a “cleaning method” (rinse one and two)
 Note the drying time in the method (e.g. 2 minutes)
 After completion of the method, read measured values
for air (OSC and d) for the cell temperature
 Wait two minutes and verify the measured values again
 If they have changed, dry manually for 30 seconds
 Wait two minutes and verify the measured values again
 If they have changed, dry manually for 30 seconds
 And so on till values become stable

34
Cleaning

Drying verification

If OSC value is decreasing (not getting stable)

 See before (increase drying time)
 Second rinse liquid not free of water (rinse with
appropriate liquid)
 Drying agent used up (replace with new one)
 Cell connectors worn out or damaged (replace
them)

OSC value is increasing (not getting stable)

 Drying agent used up, more and more
condensation from ambient moisture (replace with
new one)

35
Cleaning

Drying duration at different temperatures

Higher than room temperature

 Duration may be shortened (“auto” in method takes too long)
 Verify like before

Lower than room temperature

 Increase time by at least 50% compared to drying at room temperature
(“auto” in method is too short)
 Verify like before

36
Measuring

Common issues

 Repeatability
 Sample not homogenious or particle containing
 Air bubbles
 Calculations and conversions

37
Measuring

Repeatability within short period (hours)

 Existing instruments
- Assume first that cell is clean and dry
- Use air and local water to check
repeatability
- Within a few hours the OSC values
vary not more than +-1! To do Ideal values
Note OCS of empty cell 813962
 New installation (air)

- Cell assembly must be clean and dry Fill with water and wait for
two minutes, note OSC of
1137814

- Power up and wait for a while (e.g. 15 water

or 30 min.) before measuring for the Run cleaning method 813962
system to warm up (rinse 1 and 2, drying)
and wait two minutes,
note OSC of air
Dry manually for another 813962
minutes and wait two
further minutes, note
OSC of air

38
Measuring

Repeatability within long period (days)

 Variations of OSC values should not exceed

- +- 5 (max. 10) for air on a 5 place instrument
- +- 2 (max. 3) for water on a 5 place instrument
- +- 1 (max. 2) for both water and air on a 4 place instrument
 If values found to be o.k.
- Means cleaning is sufficient
- Drying is adequat
 If adjustment needed: only do it after values have been verified

39
Measuring

Inhomogeneous samples not ideal

 Apply short measurement time

- Long measurement times will cause sedimentations or won‘t stabilize
 Measure sample several times and take mean value
- Sample is pushed forward for every new measurement
- Chances are higher to get a representative value

40
Measuring

Air bubbles in measuring cell

 Air bubbles (or a thin air film) is a serious problem

 Even very small quantities of air cause big errors when doing density
determinations

Diameter of the air bubble [mm] Max. Measuring error caused [g/cm3]

2.00 0.000838
1.00 0.000052
0.50 0.000003

41
Measuring

Air bubbles in measuring cell

 Measure sample several times

- Sample is pushed forward for every new measurement
- Bubble will change position in the cell and influence oscillation
- Particularly useful for viscous samples with „static“ bubbles

 Use bubble check

- Bubbles are detected based on an algorithm
- Particulary useful for non-viscous samples

42
Measuring

Calculations and conversions

 Make sure to use the correct conversion table

 Typing or transcription errors?
 Use automatic conversion if possible
 Consider temperature correction (e.g. API tables)

43
Tests

Why are tests recommended?

 A test method helps to identify whether the

instrument is clean or not
 It makes sure that the equipment performs
according to specs (GSST)
 Run regular tests
- Put a reminder
- Include in series
 Use test substance close to your sample
- You can use water as test substance for example
- Also solvents can be used depending on the
application (e.g. in Petro, toluene can be used
which has additional cleaning function)
- Or certified standards

44
Adjustment

Adjustments should only be performed if

 Test methods have failed multiple times

- In spite of good cleaning practice
- In spite of repeatability of equipment has been verified successfully
 Instrument is clean and completely dry
 Stability of equipment has to be verified
 A correct air pressure value is available
 Clean and distilled water is available

45
Agenda

 What is Density and How is it Measured?

 Good Density Measurement Practice
 What is Refractive Index and How is it Measured?
 Good Refractive Index Measurement Practice

46
Refractive Index

Visualization

air water Aqu. sugar solution

 Put a rod into a glass  Add water  Add sugar to water
 Rod looks straight  Rod looks broken  Rod even more broken
Refractometry has to do with measurement of this broken angle
(angle of refraction)
47
Refractive Index

Definition

i
Vacuum
Snell‘s law
Liquid
r
sin i Angle of incidence

Refractive n =
index
sin r Angle of refraction

Refractive index is sin (angle of incidence) / sin (angle of

refraction) and is dimensionless
48
Refractive Index

Properties

 Depends on temperature
i
→ decreases with increasing temperature Vacuum
 Depends on wavelength of incident light Liquid
→ increases with increasing wavelength r
 Is conventionally measured at 589.3 nm
(D line of sodium) and a temperature of 20°C

20 at 20°C
Refractive index: n
D at D line of Na

The refractive index depends on temperature and the

wavelength of the incident light
49
Refractive Index

How to measure refractive index?

Digital Refractometers

Abbe Refractometer

Portable optical
refractometer

METTLER TOLEDO

Old New

50
Refractive Index

Portable optical refractometer

 Open lid
 Place sample on prism, close lid
 Look into refractometer and read refractive
index off built-in scale

51
Refractive Index

Portable optical refractometer

+ Easy method
+ Cheap
+ Quick measurement
- Not possible to thermostat
- Limited measuring range
- With dark samples: difficult to read result
- Operator dependent result
- No possibility to print out result
- Accuracy questionable
- Limit of error: around 0.001 (more theoretically, but
only if….)

52
Refractive Index

Abbe refractometer

 Put sample on prism

 Close prism
 Read refractive index of built-in scale
 Light source: daylight or source with defined
wavelength
 Temperature: water bath (flow through
connections available with some models), but
often used without thermostat!

53
Refractive Index

Abbe refractometer

+ Relatively cheap
+ Quick measurement
+ Possibility to measure solids (films, depending on sample)
- Limited measuring range
- With dark samples: Difficult to read result
- Operator dependent results
- Accurate temperature control very difficult
- No possibility to print out results
- Accuracy questionable
- Limit of error: around 0.001 (more theoretically, but only if….)

54
Refractive Index

Digital refractometer

 Thermostat with Peltier element (heat/cool)

 Measure temperature
 Measure critical angle of total reflection and
convert to refractive index

The sample is in contact with the measuring prism

and is irradiated by an LED. The critical angle of the
total reflection at 589.3 nm sodium D wavelength is
measured with a high-resolution sensor array. The
refractive index (nD) is calculated from this value.
To obtain highly accurate results, the sample and
measuring prism are thermostated precisely via Peltier
Technology.

55
Refractive Index

Digital refractometer

Sample Thermo conductive block

Peltier Element
heat / cool

LED

Interference filter Temperature CCD (charge

sensor coupled device)
sensor
Lenses

56
Refractive Index

Digital refractometer

+ Built-in thermostat
+ Built-in NaD light source
+ High accuracy
+ Easy to use
+ May be automated
+ Results can be printed (GLP)
+ Quick measurement
- Expensive (initial investment only) Put sample
Press start
Read result
(or fully automatic)

57
Refractive Index

Summary

Sampling Sample Operator Temperature Initial

time volume dependency control investment

58
Agenda

 What is Density and How is it Measured?

 Good Density Measurement Practice
 What is Refractive Index and How is it Measured?
 Good Refractive Index Measurement Practice

59
Good Refrac. Measurement Practice

Overview

 Sampling
 Cleaning

60
Sampling

Common issues

 Sample quantity
 Adding sample on prism
 Evaporation and hydroscopic samples

61
Sampling

Sample quantity

 Add enough sample to fill the prism

 Some light is reflected at prism/sample interface
 Some light is reflected at air/sample interface
 If enough sample is used, this light does not hit the CCD sensor
 Correct results
Air

Sample
Scattered light hits
away from CCD
Prism

62
Sampling

If not enough sample is applied

 Usage of only a few drops

 Light reflected at air/sample interface is hitting CCD sensor
 Incorrect results

Air
Sample

Scattered light
Prism hits CCD

63
Sampling

Organic solutions

 Usage of only a few drops

 Light reflected at air/sample interface is hitting CCD sensor
 Incorrect results

Air
Sample

Scattered light
Prism hits CCD

64
Sampling

How much is enough?

 Fill at least up to the ring for aqeous samples (min. 0.5 mL)
 Organic solutions need a little more, no less than 0.8 – 1.0 mL

65
Sampling

Adding sticky samples manually

 Wipe off sample with a tissue

- Leaves a thin layer on prism and in corners
 Add new sample
 Measure new sample
- Unreliable measurement – mainly contamination is measured

Contamination

66
Sampling

Adding sticky samples manually

 Wipe off sample with a tissue

- Leaves a thin layer on prism and in corners
 Apply cleaning liquid and wipe it off afterwards
- Wait a few seconds to let the solvent evaporate
 Add new sample
 Stir / scratch over prism with a plastic pipette
tip or syringe
- Removes air cushions between prism and
sample
- Homogenizes sample (dilutes contamination)
 Measure sample
- More reliable measurements – contaminations
hardly visible

67
Sampling

Evaporating or hygroscopic samples

 At least use a vapor proof lid

 Best results are achieved using flow-through conditions

68
Cleaning

Common error sources

 Wrongly selected cleaning solution

 Application of cleaning liquids

69
Cleaning

Manual cleaning of prism

 Remove old sample („waste

syringe“)
 Clean with suitable solvent a few
times
- Add solvent
- Stir using waste syringe
- Remove solvent using waste
syringe
 Wipe dry with soft tissue
 Wait 10 seconds before adding
new sample
 A second solvent (e.g. Acetone)
bears the risk of contamination

70
Cleaning

Manual cleaning of prism

 Waste syringe can be used many times

- Reduction of plastic waste as well as tissue waste
- It is recommended to label it accordingly (e.g. tape)
 Sample syringe to use
- 3 component syringe with rubber O-ring to allow dispensing speed
control only if sample is not dissolving rubber on O-ring
- Otherwise use two component syringes which are all plastic

71
Questions?

72