®

The original bilberry extract.

These statements may not comply with your country’s laws and regulations or with Reg. EC n.
1924/2006 and have not been evaluated by the Food and Drug Administration. The products are notintended to diagnose, treat, cure or prevent any disease. Marketers of finished products containing
this ingredient are responsible for ensuring compliance with the applicable legal framework.

1.

Indena:beyond quality, naturally
There is a substantial amount of evidence supporting the strong
connection between health and nutrition.

Life expectancy is continuously
increasing together with a higher
attention to quality, this becoming
a key factor for both consumers
and the food industry.
2Consumers are more and more aware of what is contained in food, theylook for safe and effective ingredients and the presence of available
evidence supporting new ingredients offered in terms of benefits andsafety of their dietary supplementation may heavily inﬂuence theirbuying decision. On the other hand, the food industry is expected to meet
consumer demands, bringing to the market safe and effective products
containing safe and certified ingredients.
Natural products in various forms have been used all over the world since
time immemorial for the treatment of pathological conditions or for healthbenefits.1 Plants represent a rich source of new active principles andbotanical extracts can be used in different markets including healthfoods and supplements. The overall quality is essential, together withclinical substantiation to guarantee the necessary safety and efﬁcacy
proﬁles.

Bombardelli E., Bombardelli V.,
Twenty years experience in thebotanical health food market,
Fitoterapia 76, 495-507, 2006.
1In western countries where an allopathic systemof medicine substituted almost completely the useof crude extracts, plant derived compounds constitute
25% of the existing drugs.

Setting aside their often unsatisfactory biological and clinicaldocumentation, many extracts today are poorly chemically characterizedwith their active constituents un identified, and their constancy andstability erratic. For these reasons, many extracts with totally different
compositions claim to possess the same therapeutic properties. It would
emerge from this scenario, that to obtain biological data, reproduciblein terms of safety and efficacy, the active ingredients must be the sameover time; they must be stable and devoid of unpredictable toxicity or sideeffects. The answer can be found only in the preparation of standardized
extracts, identifying the active constituents and controlling the unknown
substances in terms of stability and constancy. Standardization is a hardtask which should begin from the biomass obtained through cultivationor collecting the plant in the wild in line with Good Agricultural Practice(GAP); this is followed by chemical isolation and the characterization ofactive reference substances, the setting up of validated analytical methods
and finally applying Good Manufacturing Practice (GMP) to the productionof the final ingredient.
A large number of ingredients with a distinctive high quality proﬁle has
been developed by Indena, starting out from astate-of-the-art clinical design study with the aim of meeting andpossibly exceeding market expectations.

Mirtoselect® is a stunning primeexample of this.

3

2.

The precious bilberry
Many red fruits are full of healthy
antioxidants, in particularanthocyanins, and may be helpful
as an adjuvant in the preventionof several conditions.

2Nutrient U.S. Department of Agriculture,A.R.S., USDA Database for the Flavonoid
Content of Selected Foods, Release 3.0
2011.
3

Prior R.L. et al., Antioxidant Capacity

As Influenced by Total Phenolic andAnthocyanin Content, Maturity, andVariety of Vaccinium Species, J. Agric.
Food Chem. 46, 2686 (1998).
4Tutin T.G., Diapensiaceae to

Myoporaceae, Flora Europaea Vol. 3,

Anthocyanins are found naturally in a number of foods - in red wine,
certain varieties of cereals, and certain leafy and root vegetables(aubergines, cabbage, beans, onions, radishes) - but are most abundant incoloured fruits like bilberries.2
Furthermore, Prior et al.3 reported that Vaccinium myrtillus L. containhigher amounts of anthocyanins than any other berries.

Cambridge University Press, Cambridge,
1972, pp 12-13.
5

Hutchinson J., British Wild Flowers Vol.

1, David and Charles, Newton Abbot, 1972,
p 226.
6

Benigni R., Piante Medicinali - Chimica,

farmacologia e terapia, Vol. II, Inverni

Vaccinium myrtillus L. is a small edible berry which grows onsmall wiry shrubs on hillsides throughout Central and Northern
Europe.4-8 Among the 450 species9 belonging to the genus Vaccinium,
the traditional medicinal use of Vaccinium myrtillus L. has been
documented since the Middle Ages, when its fruits were recommended
for a variety of conditions, and from the 16th century the plant has beensystematically mentioned in all major herbal treatises.

della Beffa, Milano, 1962, pp 951-958.
7Hegi G., Illustrierte Flora von Mittel

Europa, Band 5/3 Teil, C. Hanser Verlag,Munchen, 1927 (Reprint 1966), pp 16671681.
8Upton R. American Herbal

Pharmacopoeia and TherapeuticCompendium – Bilberry Fruit 2001.

4

Unlike most other berries, Vaccinium myrtillus L. is extremely difficult
to grow; it does not produce clusters of berries, but single or, more
rarely, pairs of berries, in limited numbers considering the biomass ofthe plant. Bilberries are softer and juicier than most other berries, andtherefore more susceptible to damage and more difficult to transport.
Bilberries cannot be cultivated, and are therefore picked from wild plants
growing on publicly accessible lands; nor can they be processed unfrozen,
since tissue damage triggers the deglycosidation of anthocyanins, withdetrimental effects on their chemical stability.

9Mabberly D.J., The Plant-Book II Ed.,

Cambridge University Press, 1997.

A number of compounds belonging to different chemical classes havebeen isolated from the berries of Vaccinium myrtillus L., particularly
the anthocyanins (also called anthocyanosides) which have been
extensively documented in scientiﬁc literature.
In the early 1970s, Indena developed a standardized bilberry extract whose
efficacy has been extensively clinically evaluated in vascular health, withover 60 positive studies, including
at least 30 controlled or double-blind studies publishedin peer-reviewed titles.10

10Morazzoni P., Vaccinium myrtillus L.,

Fitoterapia 1996, 67, 3-29.

Bilberry is one of the richest
sources of anthocyaninsc. 0.3-0.5%

on a fresh
weight basis
RR1

The anthocyanin profile
of bilberry is combinatorial

5 anthocyaninsand 3 sugars

HOO+R2O—R3OHThe profile of all other sources
is additive and characterizedby 1-3 major compounds

53.

From bilberry
to Mirtoselect®
The freshly collected berries of Vaccinium myrtillus L. are frozen inventilated tunnels at -35°C. Once frozen, the berries are machineprocessed, separated into single berries according to size, (small berries
are removed), weighted (in order to remove stones), and any leaves
discarded.
Any red lingonberries (Vaccinium vitis-idaea) which may be present areremoved through a system called Niagara. A belt moves the berries up toa point where they fall and sensors detect any falling berries which areout of the colour range. Manual checks are also carried out before thecleaned berries are either packaged or temporarily stored in large boxes.

The process from bilberry to Mirtoselect®

12Berries
are frozen
at -35°C.

Frozen berries
are sorted by size.
Small ones
are removed.

Extraction and puriﬁcation phases
follow which take the raw bilberry
extract to a puriﬁed dry extract.

63Red ligonberries
are removed throughNiagara system.

4Extraction
and purification.

Mirtoselect®
Mirtoselect® is a standardized bilberry extract containing ≥36% ofanthocyanins, and is characterized by a very specific and consistent HPLC
profile that represents the “fingerprint” of the extract. Mirtoselect® is theauthentic bilberry extract (Vaccinium myrtillus L.) obtained exclusively
from fresh fruit harvested when ripe, between July and September.

Authentic bilberry has a very specific “ﬁngerprint”
identified through HPLC analysis11-12≥36%

Authentic bilberry contains ≥36% anthocyanins,plus the full range of the non-anthocyanin components,
critical for its efficacy11-12

Indena has developed a specific and indisputable analytical method for theidentification of bilberry anthocyanins and anthocyanidins.
The HPLC based method defines the anthocyanins content of thestandardized bilberry extract as 36% anthocyanins (equivalent to 25%
anthocyanidins by UV). It has become a standard reference for mostPharmacopoeias in western countries.13-17

11Gizzi C. et al., Bilberry extracts are notcreated equal: the role of non anthocyanin
fraction. Discovering the “dark side of theforce” in a preliminary study, European
Review for Medical and PharmacologicalSciences, 2016 Jun; 20(11):2418-24.
12Motohashi N., Occurrences, Structure,Biosynthesis and Health Benefits
based on their evidences of medicinal
phytochemicals in vegetables and fruits -The Indena extract is the moststudied bilberry ingredienton the market

Volume 5 – 2016.
13

Cassinese C., New liquid

chromatography method with ultraviolet
detection for analysis of anthocyanins andanthocyanidins in Vaccinium myrtillus
fruit dry extracts and commercial
preparations., J. AOAC Int. 2007, 90 (4),The major applications investigated so far for Mirtoselect® are in therealms of vascular health and ophthalmology (retinal health), but animalmodels and pilot clinical studies suggest a broader clinical profile foranthocyanins that encompasses memory improvement, gastrointestinal
and cardiovascular health, metabolic syndrome and obesity.
It shows consistent clinical benefits especially for eye health. Thisconsistency is the result of careful biomass collection, storage andreproducible batch to batch industrial production. Clinical findings arealso linked with an optimal tolerability profile.
Mirtoselect® is a dark red-violet powder and may be utilized in a variety ofsupplement formulations. Recommended dose: 160-320 mg/day.

911-919.
14Baj E. et al, Qualitative and quantitativeevaluation of Vaccinium myrtillus
anthocyanins by HRGC and HPLC., J.
Chromatogr. 1983, 279, 365-372.
15

European Pharmacopoeia 6th Ed.

Suppl. 6.2 Fresh bilberry Fruit Dry Extract
Refined and Standardised, 3745-3747.
16

FU Mirtillo Nero Estratto Secco

Purificato e Titolato XI Ed. Suppl. 1, 2005,1469-1472
17

USP 31–NF 26 - Second Supplement

Powdered Bilberry Extract (Identificationand Assay).

74.

Main applications
The major applications investigated so far for Mirtoselect® are in therealms of vascular health and ophthalmology (eye protection at theretina level), but epidemiological surveys, animal models and pilot
clinical studies suggest a broader clinical profile of anthocyaninsencompassing memory improvement, gastrointestinal and cardiovascular
health. In particular, preliminary trials have shown potential metabolic
syndrome and glucose metabolism management activity with the use of
Mirtoselect®, which may represent an opportunity for future applications
following further research into its use in
Eye Health.18-22

18adults., J Agric Food Chem. 2010 Apr
14;58(7):3996-4000.
19

Karlsen A., Anthocyanins inhibit nuclear

factor-kappaB activation in monocytes andreduce plasma concentrations of pro-inflammatory mediators in healthy adults. J.
Nutr. 2007, 137, 1951-1954.
20

Through the beneficial effect on vascular circulation, Mirtoselect® canimprove oxygen and blood delivery to the eye, restoring the functionalityof tear secretion. Furthermore, the free radical scavenging properties
counteract oxidative stress, one of the major risk factors of dry eye
discomfort.

Krikorian R. et al., Blueberry supple-mentation improves memory in older

Mink, P. J. Flavonoid intake and cardio-

vascular disease mortality: a prospective
study in postmenopausal women. Am. J.
Clin. Nutr. 2007, 85, 895-909.
21Campbell, F. M., et al., Decrease inplasma protein markers of oxidative stressand inflammation in overweight/obese
type 2 diabetics after supplementation with

Mirtoselect® is the only bilberry
extract, void of any known side
effect, whose properties have beenclinically and pharmacologicallydemonstrated.

bilberry extract., In: Obesity facts 2012, 5(s1) p198.
22

Hoggard N. et al., A single supplement ofa standardised bilberry (Vaccinium myrtil-lus L.) extract (36 % wet weight anthocyanins) modifies glycaemic response inindividuals with type 2 diabetes controlled
by diet and lifestyle. Journal of Nutritional
Science, 2013, vol. 2, e22, page 1 of 9.

8

Studies have shown that Mirtoselect® can:Support healthy blood sugar level21-22

Improve contrast sensitivity in retinal health23

Attenuate free radical damage
associated with school-age children24

Support eye health in retinal challenges25Improve tear secretion in dry eye26

60+

More than 60 positive studies, including at least
30 controlled or double-blind studies published inpeer-reviewed titles, have validated the efficacy
of Indena’s bilberry extract in vascular health24

New positive data has recently emerged regarding the supportive roleof Mirtoselect® in eye health, in particular dry eye conditions. The
new randomized, double-blinded, placebo-controlled clinical study 26
offers further proof of the efficacy and safety profile of Mirtoselect®,where preserving adequate tear levels and maintaining the ocularsurface healthy may possibly help in attenuating discomfort and visual
disturbance.

23

Kim E.S., Clinical Evaluation of Patientswith Nonproliferative Diabetic Retinopathy Following Medication of Anthocyanoside: Multicenter Study, J. Korean Ophthalmology Soc., 2008;49(10):1629-1633.
24

Kajimoto S. et al., Therapeutic Effect ofBlueberry Extract in School Children withPseudomyopia at the Age of Crammingfor Exam to Restore Their Vision, J. New
Rem. Clin., 2000, Vol.49 N.1, 72-79.

Today Mirtoselect® is the onlybilberry extract (Vaccinium
myrtillus L.) supported by clinicaldata on dry eye condition.

25

Mazzolani F. et al., Minerva Oftalmol

2017 June;59(2)_38-41.
26

Riva A. et al, The effect of a natural,standardized bilberry extract (Mirtoselect®) in dry eye: a randomized, double
blinded, placebo controlled trial, European Review for Medical and Pharmacological Sciences, 2017;21: 2518-2525.

9

5.

Almost half a centuryof bilberry extract
Indena has been working on bilberry extract since the early seventies.
Over the years, our interest in the stunning properties of the precious
small blue berry has risen and our standardized extract Mirtoselect® istoday not only the most extensively studied bilberry extract available, butalso the market leader and benchmark.
Indena brings to the market, to customers and to final consumers a levelof quality that few others are able to guarantee; Mirtoselect® is one of
the finest examples of how Indena pioneers the development of botanicalderived dietary ingredients, and proof of the company’s expertise ininnovation and quality.

10Milestones in the story of Mirtoselect®

19701976

1980

Indena began research intoa standardized bilberry extract,
first named Myrtocyan®
“Bilberry 25% dry extract”.

Indena registered a medicinal
specialty soon to become
the world’s best-selling OTC
bilberry product.

The Indena productincluded in dietary
supplements market
under the Mirtoselect®
brand name and used
as a benchmark for thebilberry extract market.

2004

2003

Indena’s new dedicated state-of-the-art
production facility for Mirtoselect® wasinaugurated at the Settala site.
It processes up to 26,000kg of bilberries
a day, and has an annual production
capacity of more than 45 tons of extract.

The European Scientific
Cooperative On Phytotherapy(ESCOP) published a monographon Indena’s bilberry extract
in “The Scientific Foundationfor Herbal Medicinal Products”.

2005

2006

Indena updated an analyticalmethod, already used for therelease of its first registeredproduct in Italy.14 This new
validated HPLC method forbilberry quantifies all theanthocyanins present in bothplant material and in extracts

and is able to detect
anthocyanin extracts produced
from different plant materials(raspberry, blackberry, black
currant, elderberry, etc.).
This method represents
a standard reference forthe main pharmacopoeias.

Indena filed a new logo
to promote the consistency
and quality of Mirtoselect®.
This may also be used by Indena
customers to reassure consumers
about the quality of the active
ingredients they use.

2008

2007

Widespread official welcome
for the company’s efforts against
adulterated bilberry extracts.
Indena continues to actively contributeto the collection of available bilberry
extract monographs in the European,
Italian and US pharmacopoeia.28-30

Indena published its internal methodof analysis for bilberry extract13,
applicable also to finished products,
for the benefit of the Industry.
In the same year, Indena denounced
the presence on the market offraudulent bilberry extracts andimproperly labelled finishedproducts.27 The new Mirtoselect®
web site went online.

2017
To date, almost 60 peer-reviewed
papers have been published on
Indena’s standardized bilberry extract
since the early seventies, including
at least 30 controlled or double-blindstudies published in peer-reviewed
titles. Confirmation indeed of its
quality, safety and efficacy.

27
C. Artaria, R. Pace, G. Maramaldi,
G. Appendino., “DIFFERENT
BRANDS OF BILBERRY EXTRACT – A
comparison of selected components”,
NUTRAfoods 2007 6(4) 13-18.

European Pharmacopoeia 6th
Edition – Supplement 6.2 – «Freshbilberry Fruit Dry Extract Refined andStandardised».
2829
FU XI Ed, 1st Suppl. 2005, «Mirtillo
Nero Estratto Secco Purificato e
Titolato».

USP 31–NF 26 – SecondSupplement, «Powdered Bilberry
Extract» (Identification and Assay).
301112DPR0279 - 01/2018*08/2020, trademarks of Indena S.p.A. © 2020, Indena S.p.A.

® TM