Measuring N-protein has never been that inexpensive! Moreover, we are glad to introduce our alternatives to helium as carrier gas. Never get the helium sticker shock again. This webinar focuses on laboratories within the food and feed sector.

1. Cutting Costs within Cutting Edge EAS REAGINER
Technology in N-Protein Determination
Christian Schmidt
Webinar Series

2. Agenda
Introduction1
Current Situation2
The Revolutionary EAS REGAINER® Technology3
Our Next Generation N/Protein Analyzers4
Experimental Data5

3. The elementargroup – About us
12/15/2015 3
• The technology leader in
• Elemental Analysis (EA)
• Isotope Ratio Mass Spectrometry (IRMS)
• Offers an unmatched combination of
• innovative technologies,
• tailor-made solutions and
• comprehensive support.
Webinar

4. The elementargroup – Key Markets
12/15/2015 Webinar 4
AgronomyChemicals EnvironmentalEnergy MaterialsForensics

5. Tailor-Made to Customer Needs
• Elemental analyzers optimized for different analytical tasks
• vario Series - Industry-leading versatility in elemental analysis
• rapid Series - Dedicated instruments serving today’s customer needs
• trace Series - Trace-level detection of elements
• TOC Series - Fast and easy determination of total organic carbon
• inductar Series - Dedicated instruments for the analysis of metals and inorganics
12/15/2015 Webinar 5

6. Agenda
Introduction1
Current Situation2
The Revolutionary EAS REGAINER® Technology3
Our Next Generation N/Protein Analyzers4
Experimental Data5

7. Protein Analysis – Either you have to or you want to!
• legally bound protein determination for labeling
purposes
• protein content corresponds to product
properties e.g.:
• foam formation and taste of beer
• gluten-free starch
• dough properties
• protein content = price

8. Different Methods for N-Protein Determination
• Primary methods
• Dumas
• Kjeldahl
• Secondary methods (omitted)
• e.g. NIR (require primary method for calibration purposes)

9. Kjeldahl vs. Dumas Chemistry
Kjeldahl Dumas
Sample digested in sulfuric acid and a catalyst
for 1-2 hours
Sample is introduced into a furnace at high
temperature in the presence of oxygen, in a
inert gas stream. Digestion time: < 1 min
(CHNO) + H2SO4  CO2 + SO2 + H2O + NH4
+ Formation of N2 and NOx as well as other
products of combustion
Neutralization of sulfuric acid with sodium
hydroxide, formation of ammonia gas and
distillation of ammonia
Reduction of NOx to N2, trapping or removal of
other gases, detection of N2 on a TCD detector
NaOH + NH4
+  NH3 + H2O + Na+
Ammonia is typically trapped in a boric acid
solution to form ammonium borate and titrated
with sulfuric acid.

10. Kjeldahl vs. Dumas
Kjeldahl Dumas
Entire time of analysis for one sample several
hours.
Entire time of analysis for one sample approx.
3-4 minutes
Big benefit for re-determination of urgent
and/or suspect samples.
Large quantities of concentrated acids, and a
lot of flasks to wash
No concentrated acids. No toxic substances.
Sample prep in tin foils. No flasks.
Sample size: 100 mg - many grams
Big benefit for virtually unlimited sample size
Sample size: mg - 5 g
Very low initial price. Semi-automatic analysis
with automatic distillation and titration devices
available but expensive.
Low price for fully automated unit. No add-
ons, upgrades required.

11. Cost per Test using the Kjeldahl Method
Kjeldahl Method – Cost Per Test
Amount per Test 1 Test per Day 20 Tests per Day 50 Tests per Day
NaOH (Sodium Hydroxide ) 60ml $0.66 $11.91 $28.13
H2SO4 (Concentrated Sulfuric Acid) 20 ml $0.77 $13.89 $32.80
Catalyst (Copper Sulfate) 2 tablets $1.13 $20.31 $47.96
Weighing Paper 1 sheet $0.01 $0.11 $0.26
Standard Hydrochloric Acid Solution, 0.2 N 20 ml $1.37 $24.65 $58.19
Kel-Sorb (Boric Acid buffer solution with indicator) 60 ml $0.49 $8.88 $20.96
Waste Cost 200 ml $3.60 $64.80 $153.00
Total Cost Per Day $8.03 $144.55 $341.31
Cost per Test $8.03 $7.23 $6.83
Time to Analyze Samples 3.2 hours 5.5 hours 13.5 hours
Source: www.cem.com

12. Agenda
Introduction1
Current Situation2
The Revolutionary EAS REGAINER® Technology3
Our Next Generation N/Protein Analyzers4
Experimental Data5

13. Combustion According to Dumas Method
Organic compounds consists of C, H, N, O, S, Cl …
Combustion in O2 on oxygen donor at ~ 900°C
O2, N2, NOx, CO2 CO, CH4, SO2, H2O, HCl...
Typical problem of macro analysis (50 – 5000 mg)
and greasy samples
Combustion
Sample Carousel
Blank-Free Ball Valve
CombustionTube
Combustion
Furnace
EASREDUCTOR
REGAINER
EASPost

14. Unique Post-Combustion
Organic compounds consists of C, H, N, O, S, Cl …
Combustion in O2 on oxygen donor at ~ 900°C
O2, N2, NOx, CO2 CO, CH4, SO2, H2O, HCl...
post-combustion (and H2O and halogen removal)
O2, N2, NOX, CO2, SO2,...
Combustion
Sample Carousel
Blank-Free Ball Valve
CombustionTube
Combustion
Furnace
EASREDUCTOR
REGAINER
EASPost

15. Patented EAS REGAINER® Technology
• EAS REGAINER
• Metal free oxygen binding agents
• Reacts with excess oxygen and releases reducing gases
as by-product.
• Inexpensive, metal free, non-toxic
Combustion
Sample Carousel
Blank-Free Ball Valve
CombustionTube
Combustion
Furnace
EASREDUCTOR
REGAINER
EASPost

16. Patented EAS REGAINER Technology
• EAS REDUCTOR®
• Reduction of NOx to N2
• Resulting metal oxide is reduced back to active metal by
reducing gases released from EAS REGAINER
• Binding of SO2
• Only N2 and CO2 exit the reduction tube
• N2 quantification and conversion to protein content
Combustion
Sample Carousel
Blank-Free Ball Valve
CombustionTube
Combustion
Furnace
EASREDUCTOR
REGAINER
EASPost

17. Main Cost Drivers with “Classical” Duma Analyzers
• Consumption of reduction metal to bind excess
of oxygen
• Reduction tube filling for ~300 samples
• Increase lifetime by reducing just a part of the
combustion gas at the cost of sensitivity
• Helium as carrier gas

18. Changes with EAS REGAINER Technology
• Inexpensive binding of excess oxygen using
EAS REGAINER
• EAS REDUCTOR tube for up to 2000 samples
• No expensive helium as carrier gas required

19. Proof of Principle EAS REGAINER Technology
• 2000 consecutive samples shown with a rapid N exceed (7 days, 24/7)
• All results well below required repeatability according to ISO 16634-1
0.30
0.25
0.20
0.15
0.10
0.05
0.00
Diff.N[%]
2000150010005000
sample number
required repeatability according to ISO16634- 1
soy flour
dry yeast
wheat flour
starch
Matrix Protein [%]
Soy flour 34.97
Dry yeast 45.99
Wheat flour 20.59
Starch 2.31

20. Reagent Usage of the rapid N exceed for 2000 Analyses

21. Agenda
Introduction1
Current Situation2
The Revolutionary EAS REGAINER® Technology3
Our Next Generation N/Protein Analyzers4
Experimental Data5

22. Our Next Generation N/Protein Analyzers
exceed stands for EAS REGAINER Technology

23. Targets Laboratories
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• For laboratories with limited budget
• For laboratories that needs lowest
possible cost-per-sample for Dumas
analyzer
• For laboratories that needs maximum
sample flexibility
• For high-throughput laboratories that
needs maximum laboratory efficiency

24. Sample Preparation

25. Sample Preparation
Combustion
3-Fold Drying Detection
Sample Carousel
Blank-Free Ball Valve
TCD
CombustionTube
Combustion
Furnace
EASREDUCTOR
REGAINER
EASPost
Carrier Gas: Carbon Dioxide
Combustion
Gas Separation
in Tandem Operation
Detection
Sample Carousel
Blank-Free Crucible Feeding
TCD
CombustionTube
Combustion
Furnace
EASREDUCTOR
REGAINER
EASPost
Carrier Gas: Argon (Helium Optional)
The tandem operation of two CO2 selective trap columns assures
fast analysis times down to 5 minutes.

26. Cost-per-Sample
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• Carbon dioxide as carrier gas
• EAS REDUCTOR for up to 2000 samples
• More than 300 samples/day possible
• 0.25€-per-sample including gases
• 10 years warranty on furnace and TCD cell
• Argon as carrier gas
• EAS REDUCTOR for more than 1000 samples
• 300 samples/per day possible
• 0.47€-per-sample including gases
• 10 years warranty on furnace and TCD cell
Starting at 8-10 samples/day the rapid N exceed gives a better ROI compared to Kjeldahl Analyzer

27. Agenda
Introduction1
Current Situation2
The Revolutionary EAS REGAINER® Technology3
Our Next Generation N/Protein Analyzers4
Experimental Data5

28. 0.20
0.15
0.10
0.05
0.00
repeatabilityN[%]
wheat flour wheat flour rye flour wheat buckwheat glutenfree
required repeatability according to DIN EN ISO16634- 2
grape seed
flour flour flourtype 1050type 550 type 1150 bran
carbon dioxide
rapid N exceed – Analysis of Grain
• According to DIN EN ISO 16634-2
• Average repeatability of 0.020%
• Exceed 5 times the requirement
Grain Protein [%]
Wheat flour
Type 550 11.35
Wheat flour
Type 1050 12.67
Rye flour
Type 1150 8.10
Wheat bran 15.94
Buckwheat flour 9.78
Grape seed
flour 12.38
Glutenfree flour 3.17

29. 0.20
0.15
0.10
0.05
0.00
repeatabilityN[%]
horse cow piglet ox
required repeatability according to DIN EN ISO16634- 1
pigchicken lamb rabbit quail
carbon dioxide
rapid N exceed – Analysis of Animal Feed
• According to DIN EN ISO 16634-1
• Average repeatability of 0.028%
• Exceed 3.6 times the requirement
Feed Protein [%]
Horse 18.51
Cow 18.59
Chicken 14.91
Piglet 18.79
Lamb 18.18
Rabbit 18.59
Quail 21.13
Ox 19.03
Pig 14.50

30. 0.03
0.02
0.01
0.00
repeatabilityN[%]
required repeatability according to DIN EN ISO14891
cow milk ewe's milkcamel milk goat milk soy milk
argon
rapid MAX N exceed – Analysis of Milk
• According to DIN EN ISO 14891
• Average repeatability of 0.0032%
• Exceed 4.7 times the requirement
Milk Protein [%]
Cow 3.41
Camel 2.69
Sheep 5.01
Goat 2.77
Oat 1.07
Soy 3.01

31. 0.20
0.15
0.10
0.05
0.00
repeatabilityN[%]
required repeatability according to DIN EN ISO16634- 2
wheat flour rye flour buckwheat glutenfreewheat flour wheat grape seed
type 550 type 1050 type 1150 flour flour flourbran
argon
rapid N exceed – Analysis of Grain
• According to DIN EN ISO 16634-2
• Average repeatability of 0.019%
• Exceed 5.3 times the requirement
Grain Protein [%]
Wheat flour
Type 550 11.35
Wheat flour
Type 1050 12.67
Rye flour
Type 1150 8.10
Wheat bran 15.94
Buckwheat flour 9.78
Grape seed
flour 12.38
Glutenfree flour 3.17

32. Summary
• Carbon dioxide as carrier gas
• EAS REDUCTOR for up to 2000 samples
• 0.25€-per-sample including gases
• Argon as carrier gas (Helium optional)
• EAS REDUCTOR for more than 1000 samples
• 0.47€-per-sample including gases

33. Take Home Message
• Our N/Protein Analyzers with EAS REGAINER technology are:
• The industry leading Duma-type N/Protein analyzer on the market
• A serious option for any Kjeldahl user with more than 8-10 samples/day

34. Thank you!
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