Recommended Procedures for Wine BIB Shelf-life studies

Recommended Proceduresfor Wine BIB Shelf-life studiesPerformance BIB meetings in Bristol, England24 & 25 October 2012By:Patrick Sheavitop Jean-Claude VidalSophie Vialis1

Recommended Proceduresfor Wine BIB Shelf-life studiesSummary1 st Part: Wine Shelf-life2 nd Part: Protocol construction3 rd Part: Check-listSO 2mg/L4020Conclusion01801601401201008060Total SO 2Free SO 20 1 2 3 4 5 6 7 8 9month2

1 st Part: Wine Shelf-life1.1) Why this document?Currently there would appear to be a lack of clearrecommended procedures for defining and executingwine shelf-life studies.Numerous studies conducted on the shelf-life of winesare not scientifically valid, often due to the lack ofcontrol of key variables or because the test conditionsare not representative of reality.This document is intended for anyone who would like toorder, develop, conduct or evaluate a wine shelf-lifestudy. Although the example of BIB (Bag-in-Box ®)packaging is often cited, the same recommendationsapply to all types of packaging.3

1 st Part: Wine Shelf-life/ 1.2) Definition of Wine Shelf-life1.2) Definition of Wine Shelf-lifeLength of time before the wine is considered unsuitablefor consumption.Determination of when a wine exceeds it shelf-life canbe based upon both analytical and sensorial criteria.The judgment of a wine will be more severe if severalconditions must be met to “pass” the test, ifprofessional tasters are used rather than typicalconsumers and if certain benchmarks (for example thesame wine in a glass bottle) are used.4

1 st Part: Wine Shelf-life/ 1.2) Definition of Wine Shelf-lifeThe starting point is generally date of fill and the endpoint when it is to be consumed.If the chosen end point for the study is when thepackage is 1 st opened (and not completely finished)then this should be stated.The goal is to reach or exceed a target (example:average of 9 months) while minimizing variance.Number of BIBAShelf-life (months)5

1 st Part: Wine Shelf-life/ 1.3) Identify key shelf-life parameters1.3) Identify key shelf-life parametersThose defining an experimental protocol should firstunderstand the key determinates of packaged wineshelf life.Depending upon study objectives, certain of theseparameters are to be controlled (maintained constant)and others may be purposefully manipulated(explanatory variables) to determine the impact onshelf-life (response variables).In some cases, a parameter may be controlled inbeginning but may end up being tracked over time as aresponse variable.Examples: Free SO 2 , CO 2 .6

1 st Part: Wine Shelf-life/ 1.3) Identify key shelf-life parametersKey parameters may exhibit multiple causal links.For example higher storage temperatures: Increase package O 2 permeability Diminish free SO 2 levels increase microbiological risk, etc.Sometimes there can be a trade-off betweenparameters.If, for example, a producer of organic wines decides tolower levels of SO 2 : Either accept a shorter shelf-life Or better master other parameters (such as O 2 ortemperature) to compensate for the lack of SO 2 .If , at time of fill, the level of freeSO 2 is maintained lower:FreeSO 2It would be necessary to lower:eitherTotalO 2orFor example,25 mg/LInstead of40 mg/LStorage℃In order to maintain the sameshelf-life89107116months127

1 st Part: Wine Shelf-life/ 1.3) Identify key shelf-life parametersThe main shelf-life parameters are:1.3.1) Characteristics of the wine before fillingGrape varieties, wine colour, alcohol level, acidity, initialdissolved O 2 and CO 2 levels, prior oxidativereactions, etc..1.3.2) SO 2 LevelsThe ideal initial level of free SO 2 to employ varies withwine pH, type of packaging, shelf-lifetarget, microbiological risks, legislative limits, etc..8

1 st Part: Wine Shelf-life/ 1.3) Identify key shelf-life parameters1.3.3) Microbiological controlA initially high population of microorganisms, a lack ofadequate final filtration or insufficient cleaning (CIP) ofthe filler can result in off-tastes and bloated boxes as aresult of excess CO 2 produced from the metabolism ofthe micro-organisms.1.3.4) O 2 pick-up during fillingBecause of its obvious role in oxidation, it isfundamental to measure and to minimize the change indissolved O 2 in the wine as a result of filling as well asheadspace O 2O 2O 21.3.5) O 2 permeability of the package= True ingress of O 2 through a package filled with wineduring several months of storage.O 29

1 st Part: Wine Shelf-life/ 1.3) Identify key shelf-life parameters1.3.6) Use of oxygen scavengersIt is possible to add active molecules to the packageintended to remove part of dissolved or headspace O 2 .1.3.7) Damage to the packageThis parameter can influence O 2 Transmission Rate(OTR) as well as possible leakage of the liquid.Bag, side 1 Bag, side 2Intactzone1.3.8) TemperatureAn increase in temperature greatly reduces shelf-life.Tap1.3.9) ContaminantsExample on natural corks: TCA Risk of altering sensory characteristics of wine.10

1 st Part: Wine Shelf-life/ 1.4) Shelf-life indicators1.4) Shelf-life indicators1.4.1) Chemical analysisAlthough many chemical tests can be performed, formany food products, one indicator often stands outover the others.For beer or soft drinks a key shelf-life indicator is oftenthe loss of CO 2 (> x % loss means that the beverage isno longer acceptable).For wine the key indicator generally is free SO 2 . FreeSO 2 is relatively easy to measure and compare andthere is a threshold value under which the wine isgenerally no longer protected against oxidation.11

1 st Part: Wine Shelf-life/ 1.4) Shelf-life indicators1.4.2) Sensory evaluationFor shelf-life studies, sensory evaluation is essentialbecause products can potentially “pass” the routineoenological tests and still be unsuitable for the market.It is best to use a dedicated wine tasting room with welltrained assessors and to set up a system of scoring andinformation processing so that the results can be easilyanalyzed and presented.Sensory evaluation for wine is generally based uponappearance (colour, limpidity, etc), aroma and flavourand implies the correct use of wine descriptiveterminology.12

1 st Part: Wine Shelf-life/ 1.4) Shelf-life indicators1.4.3) Microbiological testingFor many food products, this is the key shelf-lifeindicator (especially relative to Food Safety).For wine this is also important but it is often takes aback-seat to other indicators because there are farmore BIBs rejected because of oxidation than becauseof microbiological activity.13

1 st Part: Wine Shelf-life/ 1.4) Shelf-life indicators1.4.4) Physical testingThis could include, for example, weight and volumetests .In the 2004 INRA Study for Performance BIB it wasshown that BIB weight loss (due to water vapourescaping from inside the package) was a function ofstorage temperature.At 15°C storage a BIB had a weigh loss of only about 1g over a 6 month period but for every 1°C increase instorage temperature, the package lost approximatelyan additional 1 g.Still this is not very much since even with a storagetemperature of 25°C the weight loss was only about0.3% of initial weight after 6 months.14

1 st Part: Wine Shelf-life/ 1.5) “Best by” dates1.5) “Best by” datesA “Best by” (or “Best Before”) date indicates when aproduct is optimal to consume rather than a “Use by”date, which indicates when a product is no longer safeto eat. For wine, a “Use by” date is never appropriate(wine past its optimal level presents no health risk).Few countries have mandatory “Best by” dates printedon the BIB package. A few require a fill date on thepackage but leave a “Best by” date optional. Mostcountries have no requirements for wine but oftenrequire (as a result of labelling laws) to correctlyindicate a minimum durability if one is provided andalso to provide any special storage conditions (forexample temperature) which need to be observed.15

1 st Part: Wine Shelf-life/ 1.5) “Best by” datesDepending upon the country and the product, there can be several shelf-lifereference dates used with sanctions if the date has expired.LEVEL OF THESUPPLY CHAINProducer/WholesalerdistributorRetailstoresConsumerhomesREFERENCEDATEDate offillingwineLimit Date to bedelivered to theretailerLimit Date tobe sold to theconsumerBest byDateNUMBER OFMONTHS AFTERFILLINGEXAMPLES OFSANCTIONS BYRETAILERS IF DATE HASEXPIRED0 3 69monthsDeliveryrefusal, returnto supplierReturn to supplier,destruction, sales16

1 st Part: Wine Shelf-life/ 1.5) “Best by” datesThe history of the wine before filling is an important determinate of shelf-lifebut this is often not even taken into account when a “Best by” date is fixed. We willtake the example of bulk wine transport.LEVEL OF THESUPPLY CHAINGrapeProductionWinemakingWineStorageBulkwineTransportWineStorageREFERENCEDATEDate ofgrapeharvestDatewine isfinishedDate winesent inbulkDate winereceivedin bulkDate ofwinefillingEXAMPLE OFNUMBER OFMONTHS BEFOREFILLINGOXYDATIVE HISTORY OFTHE WINE BEFOREFILLING-6 - 3- 2 - 1 0sulphiting sulphiting sulphiting sulphiting sulphitingOxidation/loss of free SO 217month

1 st Part: Wine Shelf-life/ 1.5) “Best by” datesFlexitank (24000 L) is a large BIB in a 20’ metal container instead of a box. As other BIB, it is best to minimise O 2 pickup (HS & DO) and the permeability ofthe bag. The fall of free SO 2 during transport bulk wine can be compensated by addingSO 2 before filling but the magnitude of the fall will affect the quality of the wine, itsshelf life and the level of total SO 2 .mg/LFree SO 260504030Departure of Flexitank (with adding of SO 2 )Filling of BIB (with adding of SO 2 )Fall of free SO 2(during thetransport of theFlexitank)20100Arrival of FlexitankShelf-life limit for the BIB wine1 3 5 7 9 11 months18

1 st Part: Wine Shelf-life/ 1.5) “Best by” datesOnce a “Best by” date is set it becomes a contractbetween the wine supplier and its customer and impliesacceptance of having the product rejected after thisdate. It can also imply that the supplier can justify theshelf-life claimed.A “Best by” date might be reasonably fixed for a box ofindustrial pasta made with consistent ingredients anduniform processing but given the highly variable natureof wine and the many parameters that influence shelflife(before, during and after filling), it is extremelydifficult for even trained scientists to estimate wineshelf-life.19

1 st Part: Wine Shelf-life/ 1.5) “Best by” datesTo avoid discriminatory practices one would expect anindication of minimum durability (if obligatory) to applyto all wine packages.But what “Best by” date should be given for a 2012Chateau Lafite and should the wine be thrown awaywhen it has exceeded the recommended date limit?Because of the waste generated by “Best by”date, some countries are considering retaining only“Use-by” - providing a clear guide to when food is nolonger good to eat.Although some retailers relay on « Best by » dates fortraceability and stock management, the date and hourof fill (codified or not) could also serve these purposes.20

1 st Part: Wine Shelf-life/ 1.5) “Best by” datesWould the consumer be any better off with:- inaccurate BIB shelf-life dates estimates (too highor too low) printed on the box ?or- a situation where the entire supply chain insures(via good filling, storage and transport practices)that the wine stays fresh until it arrives on the tableof the consumer?Regardless of whether or not a “Best by” date printedon the package it would seem appropriate to add anindication of recommended storage temperature (forexample store at less than 23°C) because of the strongreduction of shelf-life due to high storage temperatures.21

2 nd part: protocol construction2.1) Why this part?Not easy to produce a good protocol.Establish protocol by external or internal scientists.Rigorous and thoughtful protocol building Yields results that will not be doubtedWithout offering an exhaustive list, the protocol shouldtake into account the recommendations described inthis part.22

2 nd part: protocol construction/ 2.2) Declare study objectives and intended use2.2) Declare study objectives and intended useClearly formulate study objectivesExamples: test & evaluate new package or newtechnology.Compare relative to the current option (to see if analternative solution would be worth adopting).State the intended use: internal or for a wider publicIf the study is to be used for marketing purposes thescientific requirements should be higher so as to meeteventual criticism from the competition.Why?For whom?How?23

2 nd part: protocol construction/ 2.2) Declare study objectives and intended useExample of study objectives:- Measure impact on shelf-life of various film options.- Determine influence of headspace volume or % O 2 onshelf lifeor- Optimize inert gas (such as Nitrogen) flushing settingson a BIB filling machine.150 mL 50 mL 15% O 2 5% O 2SHELF LIFE?orororSHELF LIFE?SHELF LIFE?SHELF LIFE?24

2 nd part: protocol construction/ 2.3) Explanatory variables to be modified2.3) Explanatory variables to be modifiedThese are explanatory variables that are to beintentionally manipulated so as to measure theirinfluence on specific response variables chosen toindicate wine quality.Examples: Gaspermeability, TotalO 2 , SO 2 , N 2 flushingsettings, storagetemperature, etc.An explanatory variable will be assigned large enoughdifferences in interval levels so as to provokenoticeable differences in the response variables.Example: For storage temperature, it is better toassign intervals of at least 5°C between levels than 1°CWhen the explanatory variable levels are set, themeans must then be provided to reach these targetlevels and reduce variance.Example: If CO 2 is to beset at 700 and 1000mg/L, be sure that thevariance in the wine isreduced around thesetarget levels.25

2 nd part: protocol construction/ 2.3) Explanatory variables to be modifiedIf there are 3 explanatory variables with 2 differentlevels set for each, this results in 8 distinct tests (= 2 x2 x 2 = 8)Table 1 : Example of a well balanced experimentalplan on a wine to be filled in BIBFilm Total O 2 Total SO 2 Test #1 High High 1# Levels123ExplanatoryvariablesA BC1 Low Low 21 High Low 31 Low High 42 High High 52 Low Low 62 High Low 7123ResponsevariablesX Y Z2 Low High 8Limit the number of tests (example: get rid of tests 3 and 7)26

2 nd part: protocol construction/ 2.4) Keep as constant as possible the other parameters2.4) Keep as constant as possible the other parametersParameters other than those to be intentionallymodified (the explanatory variables) Examples of parameters that may be kept constant:Packaging materials (film, tap, box)Wine (O 2 , CO 2 , SO 2 , colour, acidity, microbiologicalstability, etc.)Filling (O 2 , Vol. Headspace & wine, inert gassettings, etc.)Transport conditions(duration, temperature, humidity, vibrations, etc.)Storage conditions(temperature, humidity, light, etc.)It is important to know the normal expectedvariation of these parameters.27

2 nd part: protocol construction/ 2.5) Approach reality2.5) Approach realityReproduce as much as possible the reality of theoperational steps all along the wine supply chain.Avoid, for example, to keep samples at 10°C if the wineto be sold is stored at room temperature.If the wine in the real world is to be subjected to 8hours road transport and 1 month of maritime transportthis should be reflected, if possible, in the protocol.Even if real transport is to be preferred, often acompromise must be reached, but in this case, anysimulated transport conditions should be described andjustified.28

2 nd part: protocol construction/ 2.5) Approach realityAvoid high storage temperatures (for example ≥35°C) intended to accelerate results and shortenstudy duration. Generally it is difficult to prove that the extrapolatedconclusions would have been the same had morerealistic conditions been adopted.To be preferred:Real supply chain circuits and storagetemperatures (example: ≈ 20°C)The storage position of the BIBs could potentiallyaffect O 2 ingress. BIBs are normally stored with the tap on the bottom.29

2 nd part: protocol construction/ 2.6) Packaging materials2.6) Packaging materialsIf packaging is to be selected for a future study fillrun, preferably: use the same packaging production lot numbers know the normal variances expected with thispackaging.If the filling has already taken place and this was notdone: At least make an effort to make sure that thepackage samples are representative.Note the lot numbers and other relevant informationthat may prove to be useful in interpreting study results.30

2 nd part: protocol construction/ 2.7) Filling BIBs for the study2.7) Filling BIBs for the studyThis requires a great deal of preparation Know and note (prior to the study) thecharacteristics and performance levels of the filling line(circuits, wine flushing vol., attainable headspacevolume, O 2 pick-up, filtration & filling technology, inertgas flushing options, etc.)Make sure that the wine will not be unintentionallymodified during the study run Particularly levels of O 2 , CO 2 , SO 2 .31

2 nd part: protocol construction/ 2.8) Choice of the wine2.8) Choice of the wineA highly oxygen sensitive white wine (for example ayoung Sauvignon Blanc) is often a good choice because its more rapid oxidation tends to generatemore clearly delineated differences in less timecompared to many red wines.This choice is not however always possible ordesirable.In any event it is important that the wine selectednot already be even somewhat oxidized because it would then be difficult to differentiatebetween those oxidative characteristics already in thewine and those induced by the choice of the packagingor another explanatory variable.32

2 nd part: protocol construction/ 2.8) Choice of the wineIf the wine = an explanatory variable (to be modified) Include this in the experimental plan.1 level =1 specific wine.If the wine = a parameter to be kept constant 1 wine must remain the same for all the tests.The great heterogeneity of wines = the principalreason why it is difficult to generalize based upon agiven shelf life study.Grape varieties, appellations, years, winemakingtechniques, marketing channels, etc. variations Extrapolation to other BIB wines must alwaysproceed with caution.33

2 nd part: protocol construction/ 2.9) Control Total O 2 (headspace and dissolved O 2 )2.9) Control Total O 2 (headspace and dissolved O 2 )Set target levels for dissolved O 2 and headspace O 2since this will have an important impact onSO 2 , aromas, colour Shelf-life. Know the O 2 performances of the fillingline, preferably by a preliminary audit. Determine if the O 2 results of the audit arecompatible with study objectives and experimentalplan.Prior to taking samples for the actual study, indicate inthe protocol clearly what needs to be done (see checklist in part 3).34

2 nd part: protocol construction/ 2.9) Control Total O 2 (headspace and dissolved O 2 )Sample preparation, the calibration of instruments andmeasurements should be performed by experiencedtechnicians (not by novices).For shelf-life studies, aim at an instrument accuracy of0.1 mg / L for dissolved O 2 and 0.3% for headspace O 2A knowledge of the uncertainty of the methods is usefulfor interpretation of discrepancies in the results.Reminder: maintain the recommended stabilisationtime for dissolved O 2 when using optical technology.For a reference document on the methods of measuring total oxygen in filled BIBs, seehttp://www.b-i-b.com/bib/web/downloads/7O2SheaVidalVialisPerfBIB29nov2010EN2.pdf35

2 nd part: protocol construction/ 2.9) Control Total O 2 (headspace and dissolved O 2 )When many O 2 measurements must be taken withoptical instruments (using spots glued inside atransparent tap), it is not always possible to do sosequentially (finish a test before starting another)because it takes at least 15 minutes to stabilize thesignal for dissolved O 2 .In this case, measure headspace O 2 first for eachsample, then place the sample in a storage rack (withtap + spot down) with the other samples. Finallymeasure the dissolved O 2 for each sample as itapproaches its stabilization time.This saves time without stopping the filling (frequentstops on the chain being important sources of error).36

2 nd part: protocol construction/ 2.9) Control Total O 2 (headspace and dissolved O 2 )The important fall of free SO 2 in the 1 st month is mainly due to the totalamount of oxygen trapped in the packaging.Subsequently, the amount of O 2 available to the wine will depend very much on theO 2 permeability of the package but often the level of free SO 2 at 1 month correlateswell with 9 and 12 months. It is therefore recommended to monitor SO 2 levels at 1month in the experimental protocol, because it will help better predict how long thewine will remain protected by free SO 2 and thus estimate its shelf life.O 2Trapped intheheadspaceO 2Trapped asdissolved O 237

2 nd part: protocol construction/ 2.9) Control Total O 2 (headspace and dissolved O 2 )Examples of the importance of Total O 2 :Sauvignon Blanc, 75 cL glass bottle (storage 17 ° C)with 6.26 mg/bottle Total O 2 (headspace + DO) whenbottled, lost 44% of its free SO 2 after one month,while for the bottles with only 1.75 mg/bottle of TotalO 2 , the loss of free SO 2 was only 12% (1) .Chardonnay, 3L BIB (storage 20 ° C)with 6.30 mg/L Total O 2 when packed, lost 35% of itsfree SO 2 after one month,while for BIBs with only 3.14 mg/L of Total O 2 lostonly 19% of free SO 2(2).mg/L200150100500Total SO 2Free SO 20 1 2 3 4 5 6 7 8 9mois(1)Vidal J.C., Moutounet M., Impact of Operating Conditions During Bottling and of Technical Cork Permeability on theOxygen Content and Evolution of Bottled Sauvignon Blanc Wine, www.infowine.com 2011 3/2 1-16.(2)Boulet J.C., Vidal J.C., Deage M., Présentation des résultats d’une étude de conservation des vins en BIB, Proceedings ofthe 2004 (Barossa Valley, South Australia) Conference of Performance BIB, 2004, 91 pages38

2 nd part: protocol construction/ 2.9) Control Total O 2 (headspace and dissolved O 2 )Changes in Total O 2 and Free SO 2 , Sauvignon Blanc, 75 cL glass bottleBottles 1 = Initial Total O 2 (headspace + DO) = 6.26 mg/bottleBottles 2 = Initial Total O 2 (headspace + DO) = 1.75 mg/bottleTotal O 2 (mg/ bottle)!987654321Bottles 20Bottles 1Total O 2Bottles 1Bottles 2Free SO 2Wine in Bottles 1 no longerprotected after 9 months0 60 120 180 240 300 360 420 480 540 days35302520151050Free SO 2 (mg/L)39

2 nd part: protocol construction/ 2.10) Set SO 2 levels in the protocol2.10) Set SO 2 levels in the protocolInitial free SO 2 for BIB wines are often 35 to 55 mg/LLevels are a function of the O 2 sensitivity of the wine.Free SO 2 are often 10 to 15 mg/L higher for BIB thanfor wine in glass bottles.Sprotects2Tendency to reduce levels in BIB (thanks to a decreasein total O 2 after filling, lower storage temperatures, etc.)and some day this may reach the same levels as forwine glass bottles but the differences betweenpackages currently exist and should be taken intoaccount in the protocol.Of course, sometimes initial free SO 2 levels aredeliberately set lower in the experimental protocol(organic wines, active packaging alternatives, etc)40

2 nd part: protocol construction/ 2.11) Measure and compare wine quality2.11) Measure and compare wine qualityPart 1 provided several indicators of wine quality(physicochemical, sensory, microbiological). It isimportant to define in advance the criteria upon whichjudgments about the wine will be made.In the 1 st instance, the experimenter can rely on thecommonly accepted threshold of 10 mg/L of free SO 2below which the wine is insufficiently protected. To thiswe can add colour (A 420 , A 520 , CIE L*a*b*), and thelevels of volatile compounds (esters, higheralcohols, thiols, etc.).41

2 nd part: protocol construction/ 2.11) Measure and compare wine qualitySensory analysis is also an important evaluation tool.The protocol must identify the objectives and operatingconditions (service order, black or whiteglass, professional or amateur judges, descriptiveanalysis or tripartite test, etc.).The definition of a benchmark such as wine in a glassbottle or in BIB stored at 5 °C often lends sensiblesupport to the physicochemical and sensory evaluation.42

2 nd part: protocol construction/ 2.12) Setting the time horizon in the experimental protocol2.12) Setting the time horizon in the experimental protocolIf several types of package types are to be evaluated, itis advisable to select a time horizon that takes intoaccount the distribution requirements for each type ofpackaging. For example, for the BIB wine theserequirements rarely exceed 8 to 10 months(exceptionally up to 1 year), while for some glassbottles to be stored in a cellar for aging, several yearsmay be necessary.This implies that several study time horizons might beadopted if several types of packaging are involved. Inthis case however, a comparison between twopackages with different life expectancies should bebased only the shortest duration (9 months, forexample, if comparing glass bottle to BIB).43

2 nd part : protocol construction/ 2.13) Defining sample size2.13) Defining sample sizeNormally sample size is dependent upon the expectedvariance of the underlying population (the bigger thevariance the bigger the sample size). Often howeverthis variance is not well known in advance and also forreasons of cost, the sample size will sometimes not belarge enough to be entirely statistically valid, but thestudy can nevertheless produce useful results.The simplified rule of thumb is to have at least 3samples for each specific test and for each date ofanalysis.44

2 nd part: protocol construction/ 2.14) Experimental protocol simulation2.14) Experimental protocol simulationTo avoid oversights, it is recommended to use aspreadsheet that lists all the explanatoryvariables, types of analysis and dates to determine thenumber of samples in the study.Table 2 (next page) presents an example of a studywith five evaluation dates, a need for 3 samples(repetitions) for each analytical test(O 2 , CO 2 , SO 2 , colour) and 2 samples (repetitions) forthe sensory evaluation.From the spreadsheet used to create the experimentaldesign, it is sometimes useful to simulate results toverify that the protocol addresses the right questions.123# Levels123ResponsevariablesX Y ZExplanatoryvariablesA B123CSampleRequirementsp q r45

2 nd part: protocol construction/ 2.14) Experimental protocol simulationTable 2: Example of an experimental plan with the number of BIB required.Experimental planBIB (1) RequirementsFilmTotal FreeTotalTest # T0O 2 SO (2) Date 1 Date 2 Date 3 Date 42 BIB1 High High 1 5 5 5 5 5 251 Low Low 2 3 5 5 5 5 231 High Low 3 3 5 5 5 5 231 Low High 4 3 5 5 5 5 232 High High 5 3 5 5 5 5 232 Low Low 6 3 5 5 5 5 232 High Low 7 3 5 5 5 5 232 Low High 8 3 5 5 5 5 23(1)In this example, when 5 samples are listed, 3 are for the wine analysis and 2 are for the sensorial evaluation.(2)At T0 , the wine has not yet changed and the sensorial and colour evaluation could be performed within thesame test unit.46

2 nd part: protocol construction/ 2.15) Human resources in the experimental protocol2.15) Human resources in the experimental protocolIt is recommended to clearly indicate thenames, qualifications and experience of those whodevelop the protocol and those who take conduct fillingand measurements. A precise definition of tasks by allstakeholders serves to reduce errors andmisunderstandings.It is preferable for the accuracy and reproducibility ofmeasurements that the analyzes are performed by themost qualified people available and wheneverpossible, always by the same people.47

3 rd Part: Check-listWhy a check-list ?In the same way that a packing list before leaving on holidayhelps you think of everything, this check-list will allow youto begin a shelf-life study without forgetting somethingimportant.48

3 rd part: Check-list / 3.1) Why do this study?3.1) Why do this study?What do I want to know ? Objective of this phase: Develop the goals of the study Method: Define a short list of questions that the study is toanswerExamples:What is the impact on shelf-life of changing O 2 levels introduced duringfilling?Will packaging choice A provide a longer shelf-life than packagingchoice B?What is the “normal” shelf life of a BIB merlot wine filled on a manualfilling line in a small winery under “typical” conditions of filling andstorage?Will changing the filling machine settings for N 2 injection have an impacton shelf-life?49

3 rd part: Check-list / 3.2) Work methods and materials3.2) Work methods and materialsHow am I going to conduct the study? What material do Ineed? What measurements must be taken? Objective of this phase: Make a list of everything needed tocarry out the study.50

3 rd part: Check-list / 3.2) Work methods and materials3.2.1) Package Specifications BIB Volume: 1.5 L, 2 L, 2.25 L, 3 L, 4L, 5L, 10 L Type of BIB film (pet met, coex evoh, etc) with indication ofOTR (& test method) Type of BIB tap with indication of OTR (& test method) Type of BIB Box (type of printing, type of cardboard) Type of BIB Handle Specification of BIB bags (traceability codes, etc.) Procedure used to obtain samples for the study. Description of other benchmark packages (for example 75cL bottle with type of closure) Other (for example with or without O 2 scavengers)51

3 rd part: Check-list / 3.2) Work methods and materials3.2.2) Spécifications sur vin au départ dans la cuve de remplissage Description of the preparation of the wine and analysisconducted Stability of winemaking parameters & prior oxidative historyof wine. Appellation and/or grape varieties Sensorial evaluation (color, aromas, taste) Target level of initial DO(for example < 0. 5mg/L) Target level of initial CO 2 (for example 400 mg/L) Target level of initial SO 2 (for example 45 mg/L free and

3 rd part: Check-list / 3.2) Work methods and materials3.2.3) Target Temperatures before and during filling Target temperature of wine filled (for example 8 °C) Target temperature of filling plant (for example 23 °C) Description of temperature recording process.3.2.4) Specification of Filtration technology Model of unit Target Level of intermediate filtration (example: tangentialfiltration at 0.8 µm) Target Level of final filtration (example: 0.45 µm) Specification relative to O 2 pickup during filtration: is finalfiltration unit to be flushed with inert gas before start-up? Specification relative to the cleaning or changing of filters.53

3 rd part: Check-list / 3.2) Work methods and materials3.2.5) Specification of transport or agitation Description of agitation during the taking of samples Transport conditions: including avoiding unwantedmovement of samples, type of transport, etc.3.2.6) Specification of storage conditions for samples Where are samples to be stored? (at filling centre withsampling conducted on site or at lab conducting the analysis). How long are samples to be kept after the end of study? (forexample end of study + 1 month) Unless it is a study objective, the samples should not bestored in a contaminated storage area (free frompaints, solvents, cleaning products, etc.).54

3 rd part: Check-list / 3.2) Work methods and materials Target temperatures for storage (for example15°C, 20°C, 25°C) Acceptable temperature variations during storage (forexample + or – 1°C) Description of temperature recording process. Target humidity levels. Although this should be recorded, itis often not a parameter kept constant. Description of measures taken to protect the samples fromtheft or tampering. Other indications55

3 rd part: Check-list / 3.2) Work methods and materials3.2.7) Specification of damage to bag during filling BIB bags filled should be visually examined prior to test tomake sure that the filling and packing technology does notinflict undue damage that will potentially have an importantimpact on study results (unless the impact of this damage isan objective of the study). The filling centre should provide an estimation of the knownBIB leaker rate due to the filling and packing process since theleakers will eliminate or damage study samples. Dependingupon the anticipated leaker rate the number of samples mayhave to be modified or else another filling line used to conductthe study.56

3 rd part: Check-list / 3.2) Work methods and materials3.2.8) Specification of Filling technology Model of machine & degree of automation Vacuum (“on” or “off” + settings if on) N 2 flushing of gland (“on” or “off” + settings if “on”) N 2 flushing in tap (“on” or “off” + settings if “on”) Inert gas in buffer tank & flushing of wine circuits Line speed Technology used to place the bag in the box Description of cleaning (CIP) procedure to be used on fillingmachine3.2.9) Spécifications sur le niveau d’homogénéité lors du tirage Must the samples be as homogeneous as possible? Ifso, avoid the beginning and end of the filling run. Volume of BIB wine filled (or wine flushing of circuits) beforeany samples for measurement are taken (for example 100 L) Quantity of wine to remain in filling tank (and/or buffer tank)when final samples are taken for measurement.57

3 rd part: Check-list / 3.2) Work methods and materials3.2.10) Target Measurements right after filling Target dissolved O 2 in wine after filling (for example < 1.5mg/L) Target Headspace volume (for example < 60 mL) Target Headspace % of O 2 (for example < 10%) Target Microbiological levels Target volume of wine (nominal volume + or – 1.5%) Description of how measurements are to be taken58

3 rd part: Check-list / 3.2) Work methods and materials3.2.11) Determination of measurement types and frequencyTypes of measurement & methods and materials required Chemical analysis: O2, SO2, CO2, Acidity, etc. Sensory evaluation based upon appearance, aroma andflavour Microbiological testing (identification of types, count, etc.) Physical testing (weight, volume, etc.) Measurements of dissolved O 2 and headspace O 2 (volumeand % O 2 ) Those sampling and conducting measurements should havea minimum of 2 BIB Cone Meters & an optical or electrochemicalO 2 measurement instrument (indicate brand andprecision).59

3 rd part: Check-list / 3.2) Work methods and materialsFrequency of measurements For example the day of fill, at 3 days, 7 days 14 days, 30days, 60 days, 90 days, 120 days, 180 days, 360 days. Transport used to get the samples to a laboratoryExperimental plan List of parameters to modify (explanatory variables) Target levels for each parameter Number of repetitions (number of BIBs analysed for eachdistinct test) From this it is possible to construct the protocol. Requirements in terms of the number of litres of wine, thenumber of BIB bags or boxes required, etc.60

3 rd part: Check-list / 3.2) Work methods and materials3.2.12) Labelling of samples Samples must be clearly identified with an easy tounderstand code Food grade adhesive labels should be printed before thestudy fill run or else use a simple bar code generators andreader. Labels placed on the outside of boxes once they have beenfilled. The person conducting the measurements must insure thatthe bag always goes back into the same box. If the bag is also identified a small label can be placed onthe outside seam (after the weld) so as to not interfere with thebarrier film. If a permanent marker is used it should be (NSF orequivalent) food grade and marking are also to be made onthe outside seam. In all cases, verify that the system of labeling or marking willresist for the entire length of the studyTest BIB 1 -O 2 highBeginning of filling61

3 rd part: Check-list / 3.2) Work methods and materials3.2.13) Specification of position of tap during storage Tap in bottom of box (full contact of wine with inner part ofclosure) or top of box (possibility or headspace in contact withinner part of closure).3.2.14) Freshly opened BIB or not for each measurement BIBs freshly opened each time or progressively emptyingthe same BIB If progressively emptied, the amount poured each time andthe amount remaining3.2.15) People Names, Qualifications & Experience of those writing theprotocol , those conducting the measurements and thosewriting the results and conclusions62

3 rd part: Check-list / 3.2) Work methods and materials3.2.16) Other equipment and materials that must be supplied for the studyFor example, by the filling centre: : 2 large, clean tables, 2chairs, good lighting, 220 V electricity, etc.Measurement instruments and means of transport to the lab.63

3 rd part: Check-list / 3.3) Establish calendar with deliverables3.3) Establish calendar with deliverables Agreeing on a final protocol often takes a very long time butit is better than producing study results that are meaninglessbecause the protocol overlooked very important parameters. Progress reports may or may not be specified. It is best to build in a margin of safety since shelf life studiesare notorious for taking longer than originally anticipated.64

3 rd part: Check-list / 3.4) Estimate study costs3.4) Estimate study costs Costs may include internal costs and external costs butmost firms only expect estimated of external costs. Thisincludes the cost of the wine & packaging used as well as allexternal services. Generally outside lab costs are based uponthe specific number and type of tests. If the firm for who the study is to be performed does nothave the resources to manage the shelf life study then anoutside manager will have to be brought in. This outside service provider can, in addition to managingthe shelf-life study, train the firm’s own staff so as to developfuture in-house shelf life testing and evaluation capacity.65

3 rd part: Check-list / 3.5) Write up results and conclusions3.5) Write up results and conclusions Check that conclusions based up the results. Provide full protocol details so that experimentation can berepeatable. Project shortcomings should also be indicated in the studyreport, including those linked to the protocol or its execution. If the study is to published and make commercial claims, itshould be subject to the critical scrutiny of experts in thespecific study area. Project sponsors should be stated. If an outside research center/lab is used, normally it shouldbe they who write the conclusions rather than the projectsponsor.ConclusionsResults66

Conclusion: Think before you actTo define and conduct tests wine storage, we must firstidentify the key variables that influence shelf life as wellas the primary chemical, sensory, microbiological andphysical indicators to be used for the study.The application of good practices to better satisfy theconsumer is always to be encouraged, but it is oftendifficult to accurately predict the true shelf life of a BIBwine (or other type of package) because of themultitude and complexity of the parameters andespecially the heterogeneity of the product. Underthese conditions, indicating uniform “Best by…” datesseems difficult to justify on scientific grounds.67

Conclusion: Think before you actMuch prior thought must go into the construction andthe execution of a good protocol.With the application of these recommendations, resultswill be more reliable and can be used to better preparethe wine and improve filling & packaging technology aswell as storage and distribution practices.Long life to wine in BIB!68

Thanks!For more information, please contact:Patrick SHEAps@vitop.frvitop Sophie VIALISsvialis@inter-rhone.comJean-Claude VIDALvidaljc@supagro.inra.fr69