Exploring Attenuation. Greg Doss Wyeast Laboratories Inc. NHC 2012

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1 Exploring Attenuation Greg Doss Wyeast Laboratories Inc. NHC 2012

2 Overview General Testing Model

3 Brewing Control Panel Beginning Brewing Control Experienced Brewing Control

4 Good Beer Balancing Act Volatile Aroma Compounds Residual Carbohydrate Hop Flavor Color Hop Aroma Bitterness Malt Character Clarity Negative Flavor Compounds Ethanol Carbonation

5 It s All About Control Brewer s Goal 1. Complete control over all process variables. 2. Manipulate the variables to achieve the desired beer. Control Points in Brewing Ingredient Selection + Good Processing + Successful + Good = Fermentation Finishing Good Beer Malt Hops Water Adjuncts Misc. Milling Mash Sparge Boil Cooling Desired Culture Aeration Yeast Pitch Rate Culture Health Temperature Yeast Nutrition Conditioning Dry Hopping Clarification Kegging/Bottling 2010 NHC

6 Fermentation Mass Balance Carbohydrate Ethanol + CO g 0.5 g 0.5 g Carbohydrate Ethanol + Carbohydrate + CO2 1.00g 0.375g 0.25g.375g Apparent Attenuation Carbohydrate Carbohydrate + Ethanol + CO2 1.00g 0.25g 0.375g 0.375g Real Attenuation

7 Attenuation Apparent Attenuation (ASBC Apparent Degree of Fermentation) %AA = (OG TG) / OG x 100 Example: OG = 1.056, TG = (56-12)/56 x 100 = 78.57%

8 Phase 1 Determine Terminal BU:TG Balancing Beers Phase 2 Sensory Sweetness Factor

9 BU:TG BU TG

10 Factors Affecting Attenuation Fermentability Grist Mash Time Mash Temperature Mash ph Mash thickness Yeast ability to ferment Strain Sugar uptake Metabolism Culture health Inhibition

11 Carbohydrates/ Sugars Fructose 2% Glucose 8% Sucrose 6% Maltose 45% Maltotriose 14% Dextrins 25%

12 Experiment Goal Create a calculator to estimate terminal density Original Density x Fermentability x Strain Att. Factor = Terminal Density Strain Grist Temperature Time Ability to Fermentability Fermentability

13 Strain Forced Fermentation Each Strain Triplicate With Media Control Calculate Strain factor from results

14 Trial Forced Fermentation Protocol Prep Centrifuge and wash yeast sample 2x to produce clean yeast paste mix media 1 19 P (Light DME all from same lot ) Add 2g Saaz per L Autoclave (Sterilize) Blend to consistent starting density (15.0P) Shake 2 x 30 Seconds to achieve DO 8 ppm Pour 170 ml. control flask Inoculate with control yeast Inoculate 600ml. with 2.5g/150 ml washed yeast paste (Pitch Rate ~ 60 million - 80 million cell ml -1 ) Dispense into 3 x 150 ml. Centrifuge 25 ml. of Remaining Centrifuge control 4 Flasks per test. 3 with strain, 1 media control

15 Trial Forced Fermentation Protocol Data Recording Record starting densities with digital density meter Incubate all flasks shaken C Remove 1 ml. sample every three hours and cetrifuge. Measure density with refractometer Continue taking readings until refractometer readings stabilize for 2 hours. Remove flasks from shaker remove 25 ml. from flask Centrifuge 2 x to remove solids Measure density with digital density meter and record final density. Average readings for sample strain and calculate apparent attenuation. Record reading for control strain and calculate apparent attenuation.

16 Trial Forced Fermentation Protocol Analysis Tests are subject to fermentability of media Needs adjustment Calculate average control attenuation for all tests (75) Subtract average control from sample control to produce control factor Add control factor to strain attentuation. Sample Control Att - Average Control Att = Control Factor Control Factor + Strain Att = Final Att Example 1007 Media Control Att = Average Media Control Att = Att = = = 76.41%

17 60.00% 61.00% 62.00% 63.00% 64.00% 65.00% 66.00% 67.00% 68.00% 69.00% 70.00% 71.00% 72.00% 73.00% 74.00% 75.00% 76.00% 77.00% 78.00% 79.00% 80.00% 81.00% 82.00% 83.00% 84.00% 85.00% 86.00% 87.00% 88.00% 89.00% 90.00% % attenuation strain % Attenuation per strain Trial Forced Fermentation Analysis

18 60.00% 61.00% 62.00% 63.00% 64.00% 65.00% 66.00% 67.00% 68.00% 69.00% 70.00% 71.00% 72.00% 73.00% 74.00% 75.00% 76.00% 77.00% 78.00% 79.00% 80.00% 81.00% 82.00% 83.00% 84.00% 85.00% 86.00% 87.00% 88.00% 89.00% 90.00% % attenuation Strain % attenuation per strain Trial Forced Fermentation Analysis

19 Attenuation Analysis Calculate average Att Divide Strain Att by Average Att for Att Factor Example 1007 Att = Att = Ave Att = / = 97.88% / = %

20 Experiment Goal Create a calculator to estimate terminal density Original Density x Fermentability x Strain Att. Factor = Terminal Density Strain Grist Temperature Time Ability to Fermentability Fermentability

21 Fermentability Study Goals Quantify how variables affect fermentability of wort Manipulate one variable at a time ph Malt Grind Mash Thickness Density? Boil? Mash Temperature Grist Mash Time Variables

22 ph Affects performance of enzymes Malt Color Temperature Solution reaction Goal Used 5.2 Buffer (ph Star) Affects original and terminal density slightly No affect on attenuation rate Tested at different ph

23 Malt Grind Affects hydration and starch availability Used Monster MM3 Gap set at.045 Milled grain separately Rinsed with pils malt Wheat needed two passes

24 Mash Thickness Starch availability 1 qt/# 2.09ml/g

25 Fermentability Testing

26

27 Mash Protocol 180 g Malt ml liquor and buffer (2.09 ml/g = 1qt:#) into Stanley Calculate Strike temp based on malt temp Mix liquor and malt stir 10 seconds Cap place in water bath set at desired mash temp. 3 min. Remove from water bath and shake 10 seconds Back in water bath Record temperature at 5 min Record temp at 30 min Add 300 ml. boiling liquor to mash press Record temp at 60 min (Other times for time trial) Record 60 min Pour mash into press stir 2 min record temp. Press and remove 500g into flask Record weight. Within 1 min Microwave High to boil 1:45 Place on heat plate at standard setting. Add 1g Saaz hops Boil 30 min Place in ice bath. Record final weight

28 Fermentation Protocol Centrifuge wort 4 min to remove solids (Aseptically) Record wort weight Asepetically remove 0.5 ml. for density testing. Measure record denstiy with digital refractometer Blend media with sterile water to make P Shake 2 x 30 seconds to reach 8 ppm DO. Pour 2 x 170 ml. into flasks Add standard yeast mix Remove 20g from each flask. Centrifuge 2 x to remove solids Measur starting densities with digital density meter Incubate all flasks shaken C Remove 1 ml. sample every three hours and cetrifuge. Measure density with refractometer Continue taking readings until refractometer readings stabilize for 2 hours. Remove flasks from shaker remove 25 ml. from flask Centrifuge 2 x to remove solids Measure density with digital density meter and record final density.

29 Mash Sparge Boil

30

31

32 Experiment Goal Create a calculator to estimate terminal density Original Density x Fermentability x Strain Att. Factor = Terminal Density Strain Temperature Grist Mash Time Ability to Fermentability Fermentability

33 Temperature All Pilsner malt grist 60 min mash every 2 degrees Manipulate enzyme performance

34 Temperature Temp Trial ID ph Mash Temp minutes) Analysis Min Max Delta Ave Att. Factor 146F 146F % 99.81% 148F 148F % % 150F 150F % % 151F % % 151F 152F % % 153F 152F % % 153F 153F % % 154F 154.5F % % 155F 156F % % 156F 156.5F % 98.86% 157F 157F % 96.59% 158F 158F % 95.45% Median (83.07) Temperature factor = Median/Attenuation

35 % Attenuation Temperature Temp vs. % Attenuation % 99.00% 98.00% 97.00% 96.00% 95.00% 94.00% 93.00% 92.00% 91.00% 90.00% 89.00% 88.00% 87.00% 86.00% Median 83.02% 85.00% 84.00% 83.00% 82.00% 81.00% 80.00% 79.00% 78.00% 77.00% 76.00% 75.00% Temperature (F)

36 Experiment Goal Create a calculator to estimate terminal density Original Density x Fermentability x Strain Att. Factor = Terminal Density Strain Temperature Grist Mash Time Ability to Fermentability Fermentability

37 Grist 10 Different Malts/Adjunct 10% of grist Standard temp 151F 60 Min Mash

38 Grist Trial ID ph Mash Temp minutes) Analysis Min Max T T Ave Att Att per % Pils Pils % 0.00% Munich 10 Mun % -0.09% Wheat Wheat % -0.09% C-120 C % -0.21% C-40 C % -0.28% Carapil Cpil % -0.28% Oats Oats % -0.33% Chocolate Choc % -0.67% Roasted Barley Sucrose Roast Barley Sucrose % -0.68% % 0.40%

39 Pils Munich 10 Wheat C-120 C-40 Carapil 10 Oats Chocolate Roasted Barley Sucrose % Attenuation Grist % % Apt. Attenuation vs. Grist 95.00% 90.00% 85.00% 80.00% 75.00% 70.00% 65.00% malt

40 Calculate - Stout Malt Att 10% Att 1% % Affect Pils 83.02% Munich % Wheat 82.08% C % C % Carapil % Oats 79.72% Chocolate 76.30% Roasted Barley 76.19% Sucrose 87.04% Total OG Factor TG

41 Sucrose and C-Pils? Malt Att 10% Att 1% % Affect Pils 83.02% Munich % Wheat 82.08% C % C % Carapil % Oats 79.72% Chocolate 76.30% Roasted 76.19% Barley Sucrose 87.04% Total Malt Att 10% Att 1% % Affect Pils 83.02% Munich % Wheat 82.08% C % C % Carapil % Oats 79.72% Chocolate 76.30% Roasted Barley 76.19% Sucrose 87.04% Total OG Factor TG OG Factor TG

42 Experiment Goal Create a calculator to estimate terminal density Original Density x Fermentability x Strain Att. Factor = Terminal Density Strain Temperature Grist Mash Time Ability to Fermentability Fermentability

43 Mash Time Standard temp 100% Pils Malt min (Every 15 min)

44 Mash Time Mash Temp minutes) Analysis Trial ID ph Min Max T Ave Att Factor 45 Min 45 min % 96.59% 60 Min 60 min % % 75 Min 75 Min % % 90 Min 90 Min % %

45 % Attenuation Mash Time Time vs. % Attenuation % 99.00% 98.00% 97.00% 96.00% 95.00% 94.00% 93.00% 92.00% 91.00% 90.00% 89.00% 88.00% 87.00% 86.00% 85.00% 84.00% 83.00% 82.00% 81.00% 80.00% 79.00% 78.00% 77.00% 76.00% 75.00% Time (Min)

46 What Do We Do With This? Goal Create a calculator to estimate terminal density Original Density x Fermentability x Strain Att. Factor = Terminal Density 1. Calculate Grist Factor 2. Apply Temp Factor 3. Apply Time Factor 1. Apply Yeast Factor Fermentabililty Yeast Factor

47 Temp Factor - Stout Recipe Stout Stout Stout Mash Temp Mash Time Yeast Grist Att Temp Factor Time Factor Yeast Factor O Density Attenuation Terminal

48 Time Factor - Stout Recipe Stout Stout Stout Mash Temp Mash Time Yeast Grist Att Temp Factor Time Factor Yeast Factor O Density Attenuation Terminal

49 Yeast Factor - Stout Recipe Stout Stout Stout Mash Temp Mash Time Yeast Grist Att Temp Factor Time Factor Yeast Factor O Density Attenuation Terminal

50 Conclusion Phase 1 More testing/analysis Better way to use Attenuation Numbers Phase 2 Sweetness Balancing IBU vs. Att vs. Sweetness

51 Special Thanks Briess Malting 5 Star Chemicals - John John Palmer Tamara Logsdon

52 The End Contact Info: Greg Doss Wyeast Laboratories Inc.

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