Softwoods have a numerically large group of economically important renewable plants. Waste processing of trees mainly bark, needles are reasonable extent not recovered. The waste contains relatively high levels of phenolic compounds. Phenolic compounds are one of the main components that have a high potential in various fields of food, pharmacy, and other industries. This review focuses on the main uses of softwood bark and overviews the extraction and analytical methods used to determine phenolic bioactive compounds in this matrix. At this time, various extraction techniques are used to obtain secondary metabolites from bark mainly bio-active phenolic compounds. The amount of bioactive compounds derived from the matrix affects the: extraction conditions, choice of the solvent, particle size, content of the water and, in particular, the extraction method. Amount and nature of the isolated compounds greatly depend on the isolation; the isolation is possible to use different methods: extraction in a Soxhlet apparatus, Soxtec extraction, accelerated solvent extraction, ultrasound-assisted, supercritical fluid extraction, pressurized liquid extraction, and microwave assisted extraction. According to literature were selected nutraceuticals phenolic compounds (isolated from softwood bark):Astringin; Catechin; Epicatechin; Ellagic acid; Ferulic acid; Gallic acid; Hydroxymatairesinol; isolariciresinol; Isorhapontigenin; Isorhapontin,lariciresinol; Lariciresinol-9-p-coumarate; Methylthy mol;p-Coumaric acid; Piceatannol; Piceid; Podocarpic acid; Quercetin; Resveratrol; Sesquipinsapol B; Sinapic acid; Tannic acid; Taxifolin; Vanillic acid; Vladinol D. From this viewpoint, it is important to collect information on pharmacokinetic properties of the nutraceuti- cal phenolic substances isolated from bark according to published papers. Pharmacokinetics properties of phenolic bioactive substances extracted by different techniques such as: molecular weight, logP, AlogP, H-bond acceptor, H-bond donor, total polar surface area, atom molar refractivity, number of rotatable bond, number of atom, rotatable bond count, number of rigid bond, number of atom ring, and number of Hydrogen Bond were calculated by DruLito (Drug LiknessTool).

1. NUTRACEUTICALS AS POLYPHENOLIC
BIOACTIVE COMPOUNDS ANALYSIS OF
SOFTWOOD BARK AND THEIR POSSIBILITIES
OF INDUSTRY APPLICATIONS
Michal Jablonsky1*, Ales Haz1, Alexandra Sladkova1,
Petra Strizincova1, Veronika Majova1, Andrea Skulcova1,
Jozef Jablonsky2
Department of Wood, Pulp, and Paper, Institute of Natural and Synthetic Polymers,
Slovak University of Technology, Radlinského 9, 81237 Bratislava, Slovak Republic
2Central Military Hospital SNP Ruzomberok - Teaching Hospital, Gen. Miloša Vesela
21, 03426 Ružomberok, Slovak Republic
michal.jablonsky@stuba.sk

2. Challenge
• Biomass is a unique source of raw materials
• Simple and clean fractionation of the main
components of biomass represents a very
important step in the field of clean renewable
carbon economy
• new efficient methods or pre-treatment /
fractionation / extraction methods

3. Bark
• contains - secondary metabolites
• source of bioactive compounds
• application = food additives, cosmetics and
pharmacological or agricultural products

4. Bark – compounds and properties
• non-polar and polar solvents, such as fats,
saturated and unsaturated fatty acids, resins,
resin acids, waxes, phenolic and polyphenolic
compounds, stilbens, flavonoids, terpenoids,
alkaloids and ligands
• antioxidant, antimycotical, cytotoxic, antiviral,
antitumor, antimalarial, insecticidal,
antimutagenic, tumorigenic, antimycotic,
pharmacokinetic activities and other properties.

5. Methods of extraction
• soxhlet apparatus,
• soxtec extraction,
• accelerated solvent extraction,
• ultrasound-assisted extraction,
• supercritical fluid extraction,
• pressurized liquid extraction,
• microwave-assisted extraction,
• pressurized solvent extraction and enhanced
solvent extraction

6. • work Jablonsky et al. 2017
• 237 identified compound in softwood bark
• cytotoxic (properties of 25 identified substances),
• antioxidant (26 substances),
• fungicidal (20 substances),
• antibacterial (42 substances) effects,
• repellents (9 substances),
• antifeedants (2 substances)
• causes an inhibition of the growth (8 substances),
• increase the activity of pheromones or act
themselves as pheromones (10)
Jablonsky, M., Nosalova, J., Sladkova, A., Haz, A., Kreps, F., Valka, J., Miertus, S., Frecer,
V.,Ondrejovic, M, Sima, J. Surina, I. (2017). Valorisation of softwood bark through extraction of
utilizable chemicals. A review. Biotechnol Adv 35(6), 726-750. DOI: 10.1016/j.biotechadv.2017.07.007

7. Selected nutraceuticals polyphenolic
compounds (isolated from softwood bark):
• Astringin; Catechin; Epicatechin; Ellagic acid; Ferulic
acid; Gallic acid; Hydroxymatairesinol; Isolariciresinol;
Isorhapontigenin; Isorhapontin, Lariciresinol;
Lariciresinol-9-p-coumarate; Methylthymol; p-Coumaric
acid; Piceatannol; Piceid; Podocarpic acid; Quercetin;
Resveratrol; Sesquipinsapol B; Sinapic acid; Tannic acid;
Taxifolin; Vanillic acid; Vladinol D.

8. Pharmacokinetics properties of nutraceuticals
• molecular weight (MW)
• partition coefficient (logP)
• octanol-water partition coefficient (AlogP)
• H-bond acceptor (HBA),
• H-bond donor (HBD),
• total polar surface area (TPSA),
• atom molar refractivity (AMR),
• number of rotatable bond (nRB),
• rotatable bond count (RC),
• number of rigid bond (nRigidB),
• nHB (number of Hydrogen Bond).
Drug Likeness Tool (DruLiTo)

9. Table 1 Drug-Likeness Descriptors Calculated by
DruLiTo Application
Compound Mw logP AlogP HBA HBD TPSA AMR
nR
B
nAto
m
RC
nRigid
B
nH
B
Astringin 406.13 0.879 -1.46 9 7 160.07 108.91 5 51 3 26 16
Catechin 290.08 0.852 -0.936 6 5 110.38 81.07 1 35 3 22 11
Epicatechin 290.08 0.852 -0.936 6 5 110.38 81.07 1 35 3 22 11
Ellagic acid 302.01 1.366 -1.304 8 4 133.52 74.69 0 28 4 25 12
Ferulic acid 194.06 0.78 0.267 4 2 66.76 55.45 3 24 1 11 6
Gallic acid 170.02 0.964 -0.721 5 4 97.99 41.77 1 18 1 11 9
Hydroxymatairesinol 374.14 0.751 -0.428 7 3 105.45 104.11 6 49 3 23 10
isolariciresinol 360.16 0.231 -1.093 6 4 99.38 103.39 5 50 3 23 10
Isorhapontigenin 258.09 2.077 0.695 4 3 69.92 81.25 3 33 2 17 7
Isorhapontin 420.14 0.771 -1.395 9 6 149.07 113.95 6 54 3 26 15
lariciresinol 360.16 1.062 -0.64 6 3 88.38 103.34 6 50 3 22 9
Lariciresinol-9-p-coumarate 506.19 1.802 0.282 8 4 125.68 148.7 9 67 4 31 12
Methylthymol 164.12 2.808 1.728 1 0 9.23 54.93 2 28 1 10 1
p-Coumaric acid 164.05 0.751 0.766 3 2 57.53 48.8 2 20 1 10 5
Piceatannol 244.07 2.185 0.631 4 4 80.92 76.21 2 30 2 17 8
Piceid 390.13 0.742 -0.897 8 6 139.84 107.31 5 50 3 25 14
Podocarpic acid 274.16 3.16 0.778 3 2 57.53 79.32 1 42 3 21 5
Quercetin 302.04 1.834 -1.244 7 5 127.45 83.44 1 32 3 23 12
Resveratrol 228.08 2.048 1.194 3 3 60.69 74.61 2 29 2 16 6
Sesquipinsapol B 540.24 0.812 -1.468 9 5 138.07 155.22 12 75 4 30 14
Sinapic acid 224.07 1.2 -0.231 5 2 75.99 62.09 4 28 1 12 7
Tannic acid 1700.17 9.537 -5.356 46 25 777.98 420.15 31 174 11 101 71
Taxifolin 304.06 0.803 -1.369 7 5 127.45 81.47 1 34 3 23 12
Vanillic acid 168.04 0.508 -0.094 4 2 66.76 45.2 2 20 1 10 6
Vladinol D 374.14 0.314 -1.064 7 3 105.45 104.75 6 49 3 23 10

10. Sample
Lipinski´s
Rule of
Five
Ghose
Filter
CMC-50
Like Rule
Vebers
Rule
MDDR
Like
Rule
BBB
Likeness
Rule
Unweighted
QED
Weighted
QED
Astringin 0 1 0 0 0 0 1 1
Catechin 1 1 0 1 0 0 1 1
Epicatechin 1 1 0 1 0 0 1 1
Ellagic acid 1 1 0 1 0 0 1 1
Ferulic acid 1 1 0 1 0 0 1 1
Gallic acid 1 0 0 1 1 0 1 1
Hydroxymatai
resinol
1 1 0 1 0 0 1 1
isolariciresino
l
1 1 0 1 0 0 1 1
Isorhapontige
nin
1 1 1 1 0
1
1 1
Isorhapontin 0 1 0 0 1 0 1 1
lariciresinol 1 1 0 1 1 0 1 1
Lariciresinol-
9-p-coumarate
0 0 0 1 1 0 1 1
Methylthymol 1 1 0 1 0 1 1 1
p-Coumaric
acid
1 1 0 1 0 0 1 1
Piceatannol 1 1 1 1 0 1 1 1
Piceid 0 1 0 1 0 0 1 1
Podocarpic
acid
1 1 1 1 0 0 1 1
Quercetin 1 1 1 1 0 0 1 1
Resveratrol 1 1 0 1 0 1 1 1
Sesquipinsapo
l B
0 0 0 0 1 0 1 1
Sinapic acid 1 1 0 1 0 0 1 1
Tannic acid 0 0 0 0 1 0 0 0
Taxifolin 1 1 0 1 0 0 1 1
Vanillic acid 1 1 0 1 0 0 1 1
Vladinol D 1 1 0 1 1 0 1 1

11. Table 3 The various filters for screening
purposes evaluated by DruLiTo Application
Selected filters
Total number of
molecular filter
Total number of molecule
violated the rule
Lipinskies
Rule of Five
19
6
Ghose_Filter 21 4
CMC-50 Like
Rule
4
21
Vebers Rule 21 4
MDDR Like
Rule
7
18
BBB Likeness
Rule
4
21
Unweighted
QED
24
1
Weighted QED 24 1
All Selected
Filters
0
25

12. Determination of polyphenolic bioactive
compounds and their properties
• Various methods are used to evaluate the
content of substances isolated from plants
• no standardized methodology for identifying
these substances
• permanganate titration, colourimetry, and
Folin-Denis method and Folin-Ciocalteu method

13. • different reagents, with a different composition of the reaction
mixture, standards and analytical evaluations
• Antioxidant activity of bark extracts can be determined by different
assays such as
• DPPH (2,2-diphenyl-1-picrylhydrazyl),
• ABTS [2,2´-azinobis (3-ethyl benzothiazoline 6-sulfonate)],
• FRAP (ferric reducing antioxidant potential), ORAC (oxygen radical
absorption capacity),
• HORAC (hydroxyl radical averting capacity),
• FTC (ferric thiocyanate assay),
• TEAC (Trolox equivalent antioxidant capacity),
• CUPRAC (cupric reducing antioxidant power),
• PFRAP (potassium ferricyanide reducing power)
• and different type of HPLC, electrophoresis, fluorimetry, cyclic
voltammetry, amperometry, biamperometry and GC/MS

14. Industrial application and concluding remarks
• waste material in the forest industry is specific.
• bark waste is a rich source of natural compounds
acting as important substances in the field of
nutrition, health and medicine
• Two ways of utilization
▫ using raw material directly from bark after milling
to more environmentally friendly products
▫ utilization of metabolites (compounds)

15. THANK YOU
This work was supported by the Slovak Research and Development
Agency under the contracts No. APVV-15-0052, APVV-14-0393 and
APVV-16-0088.