In-House vs Outsourced Lab Analysis

Presentation Outline

• Precision vs. accuracy • Industry methods and data variance • Variance between labs • Bringing analysis in house • Benefits comparison

Precision and Accuracy • What is analytical precision?

– Analytical precision is the extent of how well you are able to achieve repeatable values

• What is analytical accuracy? – Analytical accuracy is the extent of how well

you are able to achieve the “true” value

Precision and Accuracy • So which one is more important?

– Although having both high precision and accuracy is ideal, having a high degree of precision provides consistency in making decisions

– Typically the accuracy aspect can be corrected by “adjusting your aim”

• I just want to be accurate in my analysis. I need to know the true value so that I can make the best decision. – Unfortunately, accuracy (the true value) can be in the eye of the

beholder. – With wet chemistry, there are many factors that go into

generating a value. The method used, the chemicals used, the scientist performing the analysis, the proficiency of the lab, the lag between sampling and running analysis, etc.

– Many different labs can provide many different numbers, but all of them can believe they are accurate!

Industry Reference Methods

• Ethanol: 51% Ebulliometer, 31% GC, 8% NIR, 10% Distillation

• RS: 69% Enzymatic, 26% Gold Coast, 5% HPLC

• MA: 97% Enzymatic, 3% CE • TA: 54% Manual, 46% Automatic • VA: 81% Cash Still, 10% Enzymatic, 3% CE,

3% GC, 3% Segmented Flow • pH: 100% pH meter

Butzke, Christian. “Improving winery lab performance.” Practical Winery & Vineyard Journal. Jan/Feb 2002. https://www.practicalwinery.com/janfeb02p7.htm

Industry Methods and Data Variance

Butzke, Christian. “Improving winery lab performance.” Practical Winery & Vineyard Journal. Jan/Feb 2002. https://www.practicalwinery.com/janfeb02p7.htm

Industry Methods and Data Variance

Industry Methods and Data Variance

Questions to Ask • Ask your lab what methods they use

– How are those methods validated – Is the chemist TTB certified

• What is the lab’s turn around time – Harvest is usually different – Is there a walk-in/rapid method available

• There are often different pricing levels based upon method and costs associated – Don’t pay $150 for an FTIR battery

• You are the customer – We couldn’t change a filter media without

notification

Changing Labs

• Pick a lab and/or method and stick with it • One very common cause of frustration is

using multiple labs or labs using multiple solutions

• If you switch for a reason, understand that a new lab may provide different readings that are not “incorrect” – You may need to rethink your baseline

In-house Analysis

• All wineries first have some basic level of wet chemistry in conjunction with outsourced labs

• Wineries then have a decision to make: – 1. Continue to use outsourced labs – 2. Invest in more advanced wet chemistry – 3. Invest in an Analytical Platform

Bringing Analysis In-house

• P assive measurement is done after the fact – Ok for some analysis such as labeling

but not for making quick decisions – Reaction time is limited

• Active measurements are taken in r eal t ime – Fast r esults – Offer s greater control

Typical Costs to Start a Lab • Ballpark for 15-100K case winery is

approximately $75-$100K for complete wet chemistry tools – 30% will be “odds and ends” that are often

overlooked • Operating costs

– Depending on methods, estimate $10-30 per battery via wet chemistry

• Labor

A Note on Micro

• Micro is often the last piece puzzle • Basic plating is not hard but does require

some attention to sterility. Figure about $5-7K for startup materials.

• Costs: – In-house plating: $0.25-$1/test – Outsourced plating: $20/test – Scorpion/PCR: $80-200/test

Outsourcing is so much easier…but you pay for that convenience

• Outside lab analysis – $100-150 for standard panel per sample

• $40-75 for limited panels or fast methods – Shipping costs – 1-3 days for results – Larger sample size – Lag time between sampling and analysis may

result in poor accuracy • Must or fermenting must extremely hard via outsourcing

– Precision is based on lab proficiency

What are the biggest concerns w/ starting a Lab

• Money ? – There are entry and advanced levels to most pieces of

equipment – Just ify costs with savings – How quickly is a P O for $30K in bacter ia or bar rels

signed vs $5K for lab equipment which may save many thousands in other costs

– What’s it worth to increase quality? • Time ?

– Look for packaged solutions and devices that are quick and easy to use

– All-in-one platforms are an instant lab in a compact footpr int

• Training needs ? – Wet chemistr y will r equire a good chemist or analytical

tech Platforms limit the need for trained personnel

Quality It is clear that intelligently increasing analysis and understanding one’s process should increase product quality What we also see is a definite trend that as bottle price goes up – so does lab sophistication and spend Of cour se a higher pr ice point product allows higher profits to be spent, but a dispropor tionate amount of spend occur s on lab analysis vs. at other winer ies

Our Experience with the OenoFoss Platform

• The average winery who purchases an OenoFoss is either; – Spending more than $15K/yr in outsourced

costs. These costs can and do creep up. – A newer facility with no current analytical tools

or dedicated personnel – Outside of CA with limited outsource

resources – Or has wet chemistry but had recent QA

turnover

Thank you, Questions?

A Review of Traditional Analytical Methods and In-house Platforms

Presentation Outline • Industry reference methods • Cost and time comparison of reference methods • Enzymatic platforms • FTIR platforms • Summary

Industry Methods • Ethanol – 51% Ebulliometer, 31% GC, 8% NIR,

10% Distillation • RS – 69% Enzymatic, 26% Gold Coast, 5%

HPLC • MA – 97% Enzymatic, 3% CE • TA – 54% Manual, 46% Automatic • VA – 81% Cash Still, 10% Enzymatic, 3% CE,

3% GC, 3% Segmented Flow • pH – 100% pH meter

Butzke, Christian. “Improving winery lab performance.” Practical Winery & Vineyard Journal. Jan/Feb 2002. https://www.practicalwinery.com/janfeb02p7.htm

Cost and Time Comparison – Reference Methods

• NIR • Measures Ethanol, Density, SG

• Investment cost $30k • 2 minutes per test • Sample Size 25-50 ml • Some Maintenance Required • Some standards required • High Degree of Precision

Cost and Time Comparison – Reference Methods

• GC • Measures Ethanol, Polyphenols • Investment cost $5-100k+ • 10 minutes per test • Sample Size 1-10 μl • Proper maintenance is required • Some consumables required • High Degree of Precision if maintained properly

Cost and Time Comparison – Reference Methods

• Distillation • Measures Ethanol • Investment cost $500 • 30 minutes per test • Sample Size 25-50 ml • Medium Degree of Precision

Cost and Time Comparison – Reference Methods

• Ebulliometer • Measures Ethanol • Investment cost $1-3k • 10 minutes per test • Sample Size 25-50 ml • Low Degree of Precision

Cost and Time Comparison – Reference Methods

• pH meter • Measures pH

• Investment cost $100-1k • 2 minutes per test • Sample Size 25-50 ml • Probe maintenance required • Consumables required

• Calibration Standards • Annual probe replacements ($200-500)

• High Degree of Precision if maintained and calibrated

Cost and Time Comparison – Reference Methods

• Titration • Measures total acidity • Autotitrator

• Investment cost $10k • 2 minutes per test • Sample Size 10-20 ml • Weekly Probe Maintenance • Consumables required • High degree of precision if maintained properly

• Manual Titration • Consumables required • 2 minutes per test • Sample Size 10-20 ml • Degree of precision dependent on operator

Cost and Time Comparison – Reference Methods

• Capillary Electrophoresis • Measures Malic, Lactic, Tartaric, Citric, VA

• Investment cost $5-15k • 40 minute calibration, 20 minutes per test • Sample Size 1-5 ml • Maintenance required • Consumables required • High degree of precision if instrument is well

maintained and calibrated

Cost and Time Comparison – Reference Methods

• Enzymatic • Measures VA, YAN, RS, Malic, Tartaric, Citric, Lactic,

Potassium, Glycerol, SO2 • Investment cost $5-50k depending on type of unit,

automation, and options • Some units more akin to wet chemistry – others are more

all-in-one platforms • 15 minutes per test • Sample Size 1-5 ml • High costs of reagent kits – Malic $3 per test, RS $4 per

test, YAN $4 per test, Lactic $5 per test, etc. • Short expiration dates on reagents / disposal concerns

Cost and Time Comparison – Reference Methods

• Gold Coast/Rebelein Titration • Measures RS • Gold coast kits are being discontinued

• Kits are approximately $300 • 15 minutes per test • Sample Size 10-20 ml • Consumables and glassware required • Low Degree of Precision

Cost and Time Comparison – Reference Methods

• Densitometer, Refractometer, Hydrometer • Measures Brix

• Investment cost $100-1k • 2 minutes per test • Sample Size – From 1 ml for refractometer to 150

mL+ for hydrometer • Typically no maintenance or consumables required • Densitometer and Refractometer – Moderate Degree

of Precision • Hydrometer – Lower Degree of Precision

Cost and Time Comparison – Reference Methods

• Manual Chromatography • Measures Malic, Lactic, Tartaric, Citric • 8 hours per assay group • Very labor intensive • Sample Size 10-20 ml • Consumables and solvents required • Degree of precision based on operator

Cost and Time Comparison – Reference Methods

• HPLC • Measures Malic, Lactic, Tartaric, Citric, polyphenolics,

RS, Ethanol • Investment cost $5-20k • 1 hour for calibration, prep, and assay • Sample Size 0.5-15 ml • Maintenance Required • Consumables Required • Precision based on proficiency of operator to interpret

data and maintain and calibrate instrument

Cost and Time Comparison – Reference Methods

• Cash Still • Measures VA

• Investment cost $1-2k • 5 minutes per test • Sample Size 10-20 ml • Consumables required • Low Degree of Precision

Cost and Time Comparison – Reference Methods

• Segmented Flow • Measures VA, SO2

• Investment cost $15-20k • 45 minute calibration, 2 minutes per sample • Sample Size 20-30 ml • Maintenance Required • Consumables Required • Degree of precision based on maintenance and

operator

Cost and Time Comparison – Reference Methods

• Ripper and Aeration Oxidation • Measures SO2 • Investment cost $1-2k • 5-15 minutes per test • Sample Size 10-20ml • Maintenance Required • Consumables Required • Degree of Precision based on operator, wine style

can interfere with results

Analytical Platforms

Enzymatic Platforms • Uses reagents to measure

a chemical reaction • Typically feature an

autosampler • Independent of

calibrations but dependent on reactions and reagents

• $15-20K unit cost and approx. $14-20/ test battery

Enzymatic Platforms • Operating

• Flow through device, sample is mixed with reagent • Some cleaning and maintenance required

• Parameters must be able to be measured through reaction • Not used for pH, ethanol, density, absorbance based

analysis (OD280) • Different quantification of “winemakers” total acidity,

YAN/FAN

Enzymatic Reagents • Prepackaged and ready for use • Reagent for each analysis • Shelf lives and storage – some need

refrigeration • Manufacturer specific

FTIR Platforms

• Single sample analyzes many parameters

• Very low sample volume • Purchase cost $25-40K for

mainstream units • No reagent costs

What’s inside the FTIR unit?

Input Output

Fourier Transform Infrared (FTIR) Spectrometry

FTIR Analysis • Secondary (indirect) method of analysis

• quantifying analytes by their absorbance of infrared light

• Vs primary (or reference) method of analysis • chemical reactions (wet chemistry) to produce

quantifiable products allowing analyte quantity to be calculated

VS

FTIR Sample is scanned by mid-infrared light using an interferometer

A digital detector receives the absorbance of infrared light by chemical compounds in the sample

Fourier mathematics transforms the digital signal into spectra

Spectra are compared to on board global calibration database of wine and must samples and predictions (data) are generated

Precision – Where FTIR Shines

• FTIR platforms provides the best precision available for wine analysis • Differences in the method used, the chemicals used,

the scientist performing the analysis, the proficiency of the lab, and the long lag between sample collection and running the analysis are eliminated

• The “human factor” that has the potential to generate so much variance is minimalized

The FOSS Calibration • The OenoFoss and FT-2 Winescans have a built

in calibration for wine products • Calibrations were developed for different matrices, i.e.

dry wine, sweet wine, and must, since each matrix looks very different to the infrared light

• Thousands of global wine samples and the most high tech methods, e.g. analysis by HPLC, were used to generate the built in calibrations

• The calibrations and the performance of the technology were validated using a selection of unknown samples

Adjusting the Calibration • Wait a minute, why is adjusting the calibration necessary?

• Adjusting the calibration is necessary to achieve best accuracy from the instrument

• Since FOSS already developed and validated the built in calibration, shouldn’t it already provide “best accuracy”?

• Although the built in validated calibration database contains numerous samples, we adjust for two good reasons • The built in calibrations contain a library of global samples, e.g.

samples from Chile, Australia, France, Italy, Spain, and the US, primarily California

• We are adjusting the calibrations to match the lab that we base our winemaking decisions on

• All labs have their own inherent biases, meaning each lab can provide a different number for any given parameter!

Adjusting the Calibration • Adjusting the calibration on the FOSS equipment is

accomplished in 4 steps • Running 10-20 samples with a wide range of chemical

makeup (including sugar and malic acids) on both the instrument and in the lab for each calibration profile (dry wine, sweet wine, must)

• Keep in mind that some lab methods are more precise than others!

• Using the Foss Integrator software to group these samples together into a “sample set”

• Entering the data from the wet lab for each sample in our sample set

• Using the Foss Integrator to adjust the slope and bias of the calibration by calculating a linear regression correlation between the FOSS and the wet lab

FOSS Reference Methods

Taken from “FOSS Application Note 24 r3”

Expected FOSS Accuracy after Adjustment - Ethanol

Taken from “FOSS Application Note 24 r3”

These are validation

samples after slope and

bias’ing – not the global calibration

Expected FOSS Accuracy after Adjustment - MA

Taken from “FOSS Application Note 24 r3”

These are validation

samples after slope and

bias’ing – not the global calibration

Pros and Cons of Software Based Calibration

• Software and calibrations are one more thing to manage and maintain

• FTIR software can be matched to a particular lab or method or changed, if desired

• More advanced FTIR platforms offer the creation of custom parameters

Platforms vs Wet Chemistry • Platforms:

• Use less wine volume • Take significantly less time, faster time to result • Not operator dependent • Less operator skill needed • Less operating cost

• Wet Chemistry • Results can be more “industry accepted”, even when

not factoring in calibration, method, etc • Can be purchased piece by piece

Thank you,

Questions?

FOSS FTIR Units and Applications at Harvest and

Fermentation

Contents • Overview of FOSS FTIR platforms

• WineScan • OenoFoss

• Applications during vinification, harvest, fermentation • Grape Health / Rot • Ripeness • Crush Pad • Fermentation

WineScan • The WineScan FTIR platform has been performing the bulk of routine wine

analysis for over 20 years (>1000 units sold) • Most outsourced analytical labs • Major university and research programs • Almost every major corporate winery (some running as many as 15

WineScan units)

WineScan

WineScan Out of the Box

WineScan • Flow through device does require regular

maintenance and consumables • Reagents are needed for SO2 piece. • Operating cost of 5-14 cents per test • Advanced software allows the creation of

custom parameters • Customers have made parameters for methanol, port

wines, grape rot, for example

OenoFoss • The OenoFoss is the latest

FTIR based wine analyzer to be introduced by FOSS.

• Nearly 1,000 units in operation

• The OenoFoss has lower entry costs than the WineScan, no operating costs, and continues to provide fast accurate results.

• Fewer analyses, no creation

Cuvette Space Motor

Opening Lid

Upper Window

Lower Window

Detector Unit IR-Detector

IR-Beam

Benefits of OenoFoss • Analytical Speed –

• 2 minutes per sample per test battery • Accurate and Repeatable Analysis –

• Results guaranteed to be within Foss specifications • Same results regardless of operator

• Uncomplicated • If you can use a pipette you can use an OenoFoss

• No Consumables or Wear Parts

Harvest and Fermentation Specific Applications for FTIR

Platforms

Grape Health • Visual inspection proven to not be accurate to eventual wine quality

when it comes to rot • Glycerol, Gluconic, Volatile Acidity, Ethanol

• Often all must be considered – a high glycerol, for example, with the other parameters low will not cause issues

• Grape soundness indicators/indexes that account for all (sometimes + glu/fru ratio)

• Typical threshold for glycerol, gluconic, ethanol 1 g/L; for VA 0.2 g/L • Studies show that visual % of grape rot is less of an impact to wine than

duration of grape rot • Glycerol and gluconic can vary by up to 400%, based on time of

infection, at the same % rot • Temperature therefore also important

Infections

• Microbes such as botrytis preferential to glucose • Changes in glucose / fructose ratio are an

indication of degree and length of infection

Grape Maturity/Ripeness • Brix should not be the only indication

• Ratio of amino acids to ammonium also an indication of ripeness in parallel with brix

• Nitrogen containing compounds critical to downstream processing and eventual wine quality • Total utilizable N does not

necessarily increase with ripening but easily utilized N from amino acids does increase

• Negative sensory characteristics attributed to unripeness at least partially due to N sources

• Amino acids important for formation of higher alcohols and esters

Phenolics • Folin C in WineScan ; OD 280 in OenoFoss • Why measure total phenols (tannins +

anthocyanins) • Determine optimum time to harvest • Aging potential • Responsible for color and taste

• Wet chemistry (Adams-Harbertson Assay) one of the longest, most difficult procedures

Analysis @ Crush Pad • Avoids totally relying on visual inspection • Quantitative and defined parameters for quality

and thus payment • Extremely accepted in EU (more co-ops)

• Instant results for each unit (bin, truck) of incoming grapes

• Allows for documentation • Suppliers strive for quality • Wet chemistry hard to

accomplish out of a lab

Fermentation Optimization • Of course you can closely monitor residual

sugar, volatile acidity, tartaric and malic acids • Closely monitor nitrogen compounds

• Addition of nutrients (or over addition) • Effectiveness of and “good stuff” in nutrients

• Correct environment for the dominance of chosen fermentation strain

• Rack the wine off lees sooner

Stuck Fermentations • Yeast assimilable nitrogen, N compounds • Independent glucose and fructose

measurements – not just total RS • Adjust temperature according to analysis • Watch VA (less than 0.6 g/L) for bacteria

infection and ability for yeast re-start • Lysozyme or RO

• React quickly

ML Fermentation • Selection of strain and accompanying nutrients • Important to monitor: acidity, pH, alcohol

• SO2 (WineScan only) • Monitor usage/completion of malic acid

• Speed up ML • Watch pace of ML fermentation before it gets stuck

The Value of Analysis • Cost pay back is often enormous

• Smaller wineries reduce outsourced testing fees • Often spend $15-25K/yr on outsourcing before bringing a lab in-house • Shipping time and costs

• Larger wineries become time efficient, reduce spend on chemicals, footprint of lab and other equipment required

• Increased sampling time frame • Increase sampling sizes

• Every barrel vs a sampling (especially barrel ferment) • Instant results

• Stuck fermentations, optimize additions, on the fly adjustments • More knowledge to make decisions !

• As an industry supplier we see the money being poorly optimized/informed costs every day

Thank you,

Questions?