LactoSens®R Rapid Test

Measuring lactose concentration in

low lactose / lactose-free dairy products

1 © 2021 Chr. Hansen. All rights reserved.

 Content

MARKET TRENDS
Rapid growth and opportunities for innovation

CHALLENGE OF CONTROLLING LACTOSE CONCENTRATION

Imprecise, slow or expensive current methods

LACTOSENS®
Benefits, principle and procedure of new method

CUSTOMER FEEDBACK AND INNOVATION LED TO

LACTOSENS®R

TECHNICAL PERSPECTIVE

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 Market trends

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 The lactose-free market is growing rapidly, creating opportunities for
innovation and product diversification

• A BOOMING MARKET GLOBAL LACTOSE-FREE LAUNCHES2

In 2018, lactose-free milk sales were €831 million with a 14% Number of products with no/low/reduced lactose
growth rate1
• BECOMING MAINSTREAM
Once unknown, lactose-free versions of even the biggest
mainstream brands are now an everyday sight in the +296%
supermarket1
• ROOM FOR PRODUCT DIVERSIFICATION
The market is dynamic and offers an extensive range of
product options, reflected in the number of new product
launches 2010 2011 2012 2013 2014 2015 2016 2017 2018

• HEALTH CONSCIOUS CONSUMERS

Over 50% of consumers occasionally choose lactose-free
foods, despite only 13% claim to be lactose intolerant2

1 New Nutrition Business 2019

2 New Nutrition Business survey of over 2,000 consumers in the US, Brazil, UK and Spain, 2018
3 Mintel 2019

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 The threshold level for lactose concentration differs
across countries
LactoSens® ensures that production lives up to all national regulations by measuring
values below the threshold levels.

THRESHOLD LEVELS IN SOME EU MEMBER STATES FOR USE OF THE TERMS LACTOSE-FREE AND
LOW-LACTOSE IN FOODS
COUNTRY LACTOSE-FREE LOW LACTOSE
Denmark 10 mg/100g* 1 g/100 g*
Estonia 10 mg/100g* 1 g/100 g*
Finland 10 mg/100g* 1 g/100 g*
Norway 10 mg/100g* 1 g/100 g*
Sweden 10 mg/100g* 1 g/100 g*
Germany 100 mg/100g* NA
Slovenia 100 mg/100g* NA
Hungary 100 mg/100g or mL* NA
Ireland No lactose or galactose present 1 g/100 g*

* Final product

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 Consumers increasingly focus on food safety and
quality
Food safety is moving to the foreground, with increasing demands for
transparency and trust building1

FROM FOLLOWING REGULATIONS… …TO ALWAYS TRANSPARENT

From a consumer perspective, food safety With distrust among consumers growing, food
has typically been in the background safety is moving to the foreground, raising
being a matter between the food industry demands for transparency and building trust
and regulators

1 Red Associates for Chr. Hansen

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 The challenge of controlling lactose-
free dairy products

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 Study design

Workload: Getting/generating number of required samples, …

Large variations in scope/requirements; scope not always so
defined

Choice of matrices: Which product types? How to get them?

There will always be more than you can test in!

Reference method and reference materials?

Spiking or analysis of original samples?

Robustness and interferences.

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 When and why to measure the
lactose content
WHEN WHY
• Documentation
Product release • Demand from retailers

• Traceability
QC sampling • Audits

• Process optimization
Monitoring of
• Increasing capacities
production process

• Quantity of lactase
Product development • Hydrolyzation time

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 LactoSens®

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 LactoSens® offers a unique set of benefits

SAMPLE PREPARATION NORDVAL VALIDATION

The fast sample preparation and easy test protocol LactoSens® is as accurate and validated against the
make LactoSens® flexible and always ready to use. reference method HPLC and is the only rapid method
which has obtained the validation.

TIME OF MEASUREMENT APPLICATION AREAS

The one minute measurement saves the customer The method is applicable to milk, yogurt, cream,
time and increases productivity and allow for fresh cheeses.
immediate product release.

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 LactoSens® measures the lactose level based on the reaction Enzyme
Lactose
between the immobilized enzyme and the lactose Glucose
Galactose

Electrons

PRE-TEST DURING TEST REACTION

The sensor with an immobilized ONLY lactose binds to the The enzyme oxidizes lactose and
enzyme is ready to measure liquid immobilized enzyme - generates electrons that are picked up
with a mix of sugars sugars like glucose and and measured by the biochip and the
galactose do not interfere reader

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 LactoSens® is an established method across most of the world

WE ARE PRESENT IN 50+ COUNTRIES

• Over 150 customers are
currently using LactoSens® world
wide

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 LactoSens® procedure: From getting ready to using data

GETTING READY TO USE RUNNING LACTOSENS® TEST USING DATA

LACTOSENS®

LactoSens®R
Reader Kit

Computer/
software setup
Analyse (.xls)/
Sample preparation Run test(s)
export data (.csv)
LactoSens®R
Assay Kit

Equipment needed
but not included

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 Sample preparation step-by-step

LACTOSENS®R SAMPLE PREPARATION

RELEVANT DILUTION

LACTOSENS®R AND
LACTOSENS® 0.02%
LACTOSENS®R FOR NOLA® FIT

1+4 dilution 1+1 dilution

Liquid product
e.g. 1 ml milk + e.g. 1.2 ml sample +
e.g. milk, cream
4 ml buffer 1.2 ml buffer
Solid product 1+4 dilution 1+1 dilution
e.g. yogurt, e.g. 1 g yogurt + e.g. 0.91 g curd +
curd, cheese 4 ml buffer 0.91 ml buffer

REMEMBER
The quality of the test is never better than the quality of
the sample – make sure the sample is representative,
e.g. from a well-mixed and homogenous product
volume

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 Customer feedback and innovation
led to second generation with
improved features - LactoSens®R

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 LactoSens® customers are very satisfied

POSITIVE FEEDBACK CHALLENGES EXPERIENCED

Customer A Customer A
• ”Now we have that fast method to positively release our • ”The durability of the adaptor which you insert the sensors in.” This
lactose-free products with, so we are certain that we do and other customers experienced issues with the robustness of the
not release anything to the market which will cause adaptor.
people stomach aches. It feels good.” Customer B
Customer B • “We have some brown [Chocolate and coffee] products which I gave
• “It was really important for us that we could measure the up on a long time ago, we knew it did not work on LactoSens”.
mixed sample fast before we continue with the Customer D
production instead of measuring on the finished product
which we have packed and spent time on.” • “A certificate extended to more matrices would be of great help”
Customer C Customer E
• “We are spared from cost of analysis and shipping, we • “Sample application and pressing start in time is confusing at the
save a lot of time on stock… timewise, a huge difference.” beginning”

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 Based on customer feedback, the LactoSens®R Reader has been
developed with improved specifications

ADAPTOR SAMPLE DETECTION CALIBRATION MODULE

The adaptor has been improved and When a liquid is detected the Reader can be checked regularly and
integrated into the reader for a more software automatically counts down software generates calibration report
stable sensor connection for the measurement to begin for documentation

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 The calibration module works with both the new and the old reader

HOW TO CALIBRATE THE READER

• Calibration requires the new software

version 2.0

• Select reader calibration in the program

menu and follow instructions

• The reader automatically generates a PDF

for documentation

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 Feedback inspired the development of the LactoSens®R Assay Kit

INCREASED VERSATILITY RESILIENT AND ROBUST

LactoSens®R is applicable for a vast range of dairy Interference-free (Vitamins, sugars, ingredients).
products. It works with white mass or finished Highest reproducibility with an RSDR < 10%.
product (e.g. coffee, vanilla, fruit).

DETECTION LIMIT OF 0.008% EXTERNALLY CERTIFIED

Quantitative results with high accuracy at and below LactoSens®R is certified by NordVal in an extended
the threshold limit of 0.01%. range of products.

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 LactoSens®R shows very high precision on previously difficult samples
This opens a much wider range of applications for use with LactoSens®R

LACTOSENS®R VS HPLC LactoSens®R

% lactose HPLC The application range of
0.06 LactoSens®R has been
expanded. The following
0.05 ingredients no longer
experience
0.04 interferences:
• Coffee
0.03 • Chocolate
• Most fruit preps
0.02

0.01

0.00
Fruit yogurt Milk coffee Chocolate milk High protein Flavored
milk drink dairy drink

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 Test performance: White yogurt and fruit preps
LactoSens®R performs perfectly equally to the HPLC reference method and avoids previous interference issues

LACTOSENS® 0.01% VS HPLC LactoSens® 0.01% LACTOSENS®R VS HPLC LactoSens®R [%]

External HPLC [%] External HPLC [%]
% lactose % lactose

0.030 0.030

0.025 0.025

0.020 0.020

0.015 0.015

0.010 0.010

0.005 0.005

0.000 0.000
White yogurt Cherry fruit Banana fruit Peach apricot Strawberry White yogurt Cherry fruit Banana fruit Peach apricot Strawberry
mass 0.1% fat prep 18% prep 17% fruit prep 17% fruit prep 17% mass 0.1% fat prep 18% prep 17% fruit prep 17% fruit prep 17%

”< 0.01” for HPLC and LactoSens® 0.01% both shown as 0.008 for comparative reasons. ”< 0.008” for LactoSens®R also shown as 0.008

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 Test performance: Difficult samples (coffee, chocolate, …)
LactoSens®R performs perfectly equally to the HPLC reference method and avoids previous interference issues

LACTOSENS® 0.01% VS HPLC LACTOSENS®R VS HPLC

LactoSens® 0.01% LactoSens®R [%]
% lactose External HPLC % lactose External HPLC

0.12 0.12

0.10 0.10

0.08 0.08

0.06 0.06

0.04 0.04

0.02 0.02

0.00 0.00
Milk coffee <0.1% High protein High protein Buttermilk papaya Milk coffee <0.1% High protein High protein Buttermilk papaya
pudding pudding mango <0.1% pudding pudding mango <0.1%
chocolate 1.5% coconut 1.5% chocolate 1.5% coconut 1.5%

”< 0.01” for HPLC and LactoSens® 0.01% both shown as 0.008 for comparative reasons. ”< 0.008” for LactoSens®R also shown as 0.008

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 LactoSens®R shows very high precision and accuracy
with recovery rates close to 100%
RECOVERY PERFORMANCE EXPLANATION
Tests run by 3 users over 3 days. Deviation from
100% recovery (0% = center = perfectly accurate)
Recovery is a measure
40% of the percentage
35%
identified of a known
30%
25% amount of lactose
20% tested.
15%
10% • High accuracy: Results
5% in the series fall
0%
within ±10%
• High precision: All the
results are rather
tightly grouped with
no obvious outliers

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 LactoSens®R is certified by NordVal

THE COMPANY BEHIND THE CERTIFICATE

• NordVal is an independent The certificate 048/7 March 2019 is a comparison of LactoSens®R against the HPLC
third-party that evaluates and reference method at two external ISO 17025 accredited labs (Austrian Agency for
reviews the quality Health and Food Safety and LZBW Wangen, Germany)
characteristics of analytical
methods NordVal ensures high quality by measuring:
• Ruggedness: temperature, fat, cross reactivities (sugars, vitamins, flavors and
• A NordVal certificate fruit preps)
documents that a new method • Method performance characteristics: field of application, concentration range,
gives results that correspond LOD and LOQ
to the reference method for
the analysis • Internal reproducibility
• Storage stability, batch to batch variation

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 Documentation available for the different applications

LS LactoSens® 0.02% LSR LactoSens®R LSRN LactoSens®R for NOLA® Fit1

APPLICATION NOTE NORDVAL CERTIFIED

Milk LS LSR LSRN LSR
Cream LS LSR LSRN LSR
Yogurt LS LSR LSRN LSR
Sour Cream LSR LSR
Mozzarella LS LSR
Cottage cheese LS LSR LSR
Curd LSR LSR
Fresh cheese LSR
Hard cheese LSR

1 LactoSens®R for NOLA® Fit will be certified by NordVal in 2019

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 Sample preparation process for different groups of dairy products

PRODUCT GROUP SAMPLEa SHREDb HOMOGENIZEc DILUTEd MIXe CUT TIPf

Milk and cream P

Yogurt and sour cream W
Cottage, curd and
fresh cheese W
Mozzarella and
hard cheese W
a pipette (P) or weigh out on balance (W) d with appropriate volume of dilution buffer
b use a knife or grater e vortex or shake vigorously
c with use of mortar, stomacher bag or equivalent f to avoid clogging of pipette

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 A full range of dairy products have been successfully tested with
LactoSens®R
PRODUCT GROUP
These groups are directly mentioned in the NordVal certificate
Milk and cream
• Liquid dairy products that can be pipetted, diluted and tested as milk and cream can be considered
similar and will test in a similar manner
Yogurt and sour cream • Products that can be weighed out like yogurt and then diluted and tested can be considered to test in
the same manner
Cottage, curd and
fresh cheese

Mozzarella and Products similar to mozzarella and hard cheese were not included in the certification
hard cheese • Testing these with LactoSens®R products will be covered by application notes and data in those

The NordVal method covers groups characterized by their physical characteristics and sample preparation steps (see previous
slide). Dairy products that can be tested similarly can be considered as covered by the certification.

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 Choose the right LactoSens® product based on lactase and intended
lactose concentration

Concentration 0.01% 0.02% 0.2% 1%

Both the existing LactoSens® and new
LactoSens®R reader can be used with all tests.
Ha-lactase™ LactoSens® 0.02% Prod No.714978 The new reader offers improved features such
(neutral as drop detection for increased sample-to-
yeast sample consistency. To run LactoSens®R tests,
Prod No.
lactases) LactoSens®R software version 2.0 is required.
719766

LactoSens® Reader (714977)1

NOLA® Fit LactoSens®R for Prod No. LactoSens®R Reader Kit (719604)
NOLA® Fit 719602

1 Under deletion March 2019

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 Thank you

DKCHBE@CHR-HANSEN DKOMA@CHR-HANSEN DKSOMA@CHR-HANSEN

Christian Bendix Jensen, Ole Madsen, Sofie Martinussen,
Director Test kits Equipment Commercial Development Manager Product Management Assistant

30 © 2021 Chr. Hansen. All rights reserved.

 Thank you

FOLLOW CHR. HANSEN

DISCLAIMER. The information contained herein is presented in good faith and is, to the best of our knowledge and belief, true and reliable. It is offered solely for your consideration, testing and evaluation, and it is subject to change without prior
and further notice unless otherwise required by law or agreed upon in writing. There is no warranty being extended as to its accuracy, completeness, correctness, non-infringement, merchantability or fitness for a particular purpose. To the best of
our knowledge and belief, the product(s) mentioned herein do(es) not infringe the intellectual property rights of any third party. The product(s) may be covered by pending or issued patents, registered or unregistered trademarks or similar
intellectual property rights. All rights reserved.
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 Appendix

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 LactoSens is the result of more than 10 years of research

HISTORY OF DIRECTSENS GMBH

• 10 years of fundamental research

• Founded in June 2013, privately held
• Biosensors for food- and med-tech
• 2 patents protect core technology platform
• ISO 9001 certified sensor production and kit
assembly in Vienna
• 12 employees (8 R&D and product
innovation)

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