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Maura Ferri, Jürgen Graen-Heedfeld, Karlheinz Bretz, Fabien Guillon, Elisa Michelini, Maria Maddalena Calabretta, Matteo Lamborghini, Nicolò Gruarin, Aldo Roda, Axel Kraft, Annalisa Tassoni
Peptide Fractions Obtained from Rice By-Products by Means of an Environment-Friendly Process Show In Vitro Health-Related Bioactivities
www.plosone.org
www.plosone.org
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1.4683 0 Td
(development/optimisation)Tj
10.8283 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4684 0 Td
(diges-)Tj
-35.2004 -1.2982 Td
(tion)Tj
1.8425 0 Td
(process)Tj
3.1804 0 Td
(conditions)Tj
4.4787 0 Td
(were)Tj
2.1203 0 Td
(performed)Tj
4.4674 0 Td
(in)Tj
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(15)Tj
1.1735 0 Td
(mL)Tj
1.5761 0 Td
(final)Tj
2.0012 0 Td
(volume)Tj
3.2031 0 Td
(reactions.)Tj
4.0762 0 Td
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3.549 0 Td
(five)Tj
1.6554 0 Td
(differ-)Tj
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(commercial)Tj
4.9549 0 Td
(proteases)Tj
3.9174 0 Td
(were)Tj
2.1204 0 Td
(tested)Tj
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(at)Tj
0.9525 0 Td
(1)Tj
0.6973 0 Td
(U/g)Tj
1.7518 0 Td
(\(1)Tj
1.0318 0 Td
(enzyme)Tj
3.3051 0 Td
(unit)Tj
1.8652 0 Td
(per)Tj
1.5307 0 Td
(gram)Tj
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(of)Tj
1.0148 0 Td
(protein\))Tj
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(enzyme/)Tj
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(substrate)Tj
3.7984 0 Td
(ratio)Tj
2.0976 0 Td
(\(E/S\),)Tj
2.5001 0 Td
(and)Tj
1.7348 0 Td
(2)Tj
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(hour)Tj
2.1543 0 Td
(incubation)Tj
4.5524 0 Td
(time)Tj
2.0239 0 Td
(at)Tj
0.9524 0 Td
(the)Tj
1.4741 0 Td
(following)Tj
3.9798 0 Td
(pH)Tj
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(and)Tj
1.7291 0 Td
(temperature)Tj
5.1307 0 Td
(optimal)Tj
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(enzyme)Tj
3.2995 0 Td
(conditions)Tj
4.4787 0 Td
(according)Tj
4.1895 0 Td
(to)Tj
1.0205 0 Td
(literature)Tj
3.8835 0 Td
(or)Tj
1.0998 0 Td
(to)Tj
1.0261 0 Td
(manufacturer's)Tj
6.2589 0 Td
(instructions:)Tj
5.227 0 Td
(Neutrase)Tj
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Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 540.6236 590.4 cm
BT
/F15 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(1)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 550.2614 585.9212 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(0.8L)Tj
-35.0246 -1.2982 Td
(\(neu,)Tj
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(pH)Tj
1.5024 0 Td
(7,)Tj
0.924 0 Td
(50C,)Tj
2.4492 0 Td
(Novozymes)Tj
4.8982 0 Td
(A/S,)Tj
1.9332 0 Td
(Denmark\),)Tj
4.6148 0 Td
(papain)Tj
2.9423 0 Td
(\(pap,)Tj
2.2507 0 Td
(pH)Tj
1.508 0 Td
(6.5,)Tj
1.6215 0 Td
(50C,)Tj
2.4491 0 Td
(Sigma-Aldrich,)Tj
-29.3781 -1.2983 Td
(Milan,)Tj
2.8346 0 Td
(Italy\),)Tj
2.5569 0 Td
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ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 286.4692 564.3779 cm
BT
/F15 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(1)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 296.107 559.9559 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(2.4L)Tj
1.9332 0 Td
(\(alc,)Tj
1.8935 0 Td
(pH)Tj
1.5024 0 Td
(7,)Tj
0.924 0 Td
(60C,)Tj
2.4492 0 Td
(Novozymes)Tj
4.8982 0 Td
(A/S,)Tj
1.9332 0 Td
(Denmark\),)Tj
4.6148 0 Td
(Protamex)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 536.3716 564.3779 cm
BT
/F15 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(1)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 546.0661 559.9559 cm
BT
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1 TL
-0.004 Tc
0 0 Td
(\(pro,)Tj
-34.6051 -1.3039 Td
(pH)Tj
1.5024 0 Td
(8,)Tj
0.9241 0 Td
(60C,)Tj
2.4491 0 Td
(Novozymes)Tj
4.8982 0 Td
(A/S,)Tj
1.9332 0 Td
(Denmark\),)Tj
4.6148 0 Td
(Flavourzyme)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 415.0488 551.3952 cm
BT
/F15 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(1)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 424.6866 546.9165 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(500L)Tj
2.1883 0 Td
(\(flav,)Tj
2.2053 0 Td
(pH)Tj
1.508 0 Td
(7,)Tj
0.9185 0 Td
(60C,)Tj
2.4548 0 Td
(Novozymes)Tj
-31.7421 -1.2982 Td
(A/S,)Tj
1.9332 0 Td
(Denmark\).)Tj
4.6148 0 Td
(The)Tj
1.7858 0 Td
(digestions)Tj
4.2463 0 Td
(were)Tj
2.1203 0 Td
(stopped)Tj
3.3732 0 Td
(by)Tj
1.1792 0 Td
(boiling)Tj
3.0274 0 Td
(the)Tj
1.4682 0 Td
(samples)Tj
3.3845 0 Td
(in)Tj
1.0375 0 Td
(a)Tj
0.652 0 Td
(water)Tj
2.4321 0 Td
(bath)Tj
1.9899 0 Td
(for)Tj
1.3833 0 Td
(10)Tj
1.1735 0 Td
(min)Tj
-35.8012 -1.2983 Td
(to)Tj
1.0261 0 Td
(inactivate)Tj
4.0649 0 Td
(the)Tj
1.474 0 Td
(enzymes.)Tj
3.8891 0 Td
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4.2179 0 Td
(optimisation)Tj
5.3121 0 Td
(experiments)Tj
5.1363 0 Td
(mainly)Tj
2.9933 0 Td
(focused)Tj
3.2769 0 Td
(on)Tj
1.2755 0 Td
(E/S)Tj
1.5931 0 Td
(ratio)Tj
-34.2592 -1.3039 Td
(testing)Tj
2.88 0 Td
(0.05,)Tj
2.0976 0 Td
(0.5,)Tj
1.6214 0 Td
(1,)Tj
0.9241 0 Td
(2,)Tj
0.9184 0 Td
(and)Tj
1.7291 0 Td
(10)Tj
1.1736 0 Td
(U/g.)Tj
1.9785 0 Td
(In)Tj
1.1055 0 Td
(all)Tj
1.1452 0 Td
(experiments,)Tj
5.3688 0 Td
(the)Tj
1.4684 0 Td
(final)Tj
2.0011 0 Td
(pH)Tj
1.508 0 Td
(was)Tj
1.6951 0 Td
(checked)Tj
3.4696 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4684 0 Td
(total)Tj
-34.2819 -1.2983 Td
(digestates)Tj
4.0762 0 Td
(and)Tj
1.7348 0 Td
(the)Tj
1.4683 0 Td
(supernatant)Tj
4.9833 0 Td
(fractions)Tj
3.719 0 Td
(\(obtained)Tj
4.0705 0 Td
(after)Tj
2.0353 0 Td
(centrifugation)Tj
5.8903 0 Td
(for)Tj
1.3833 0 Td
(10)Tj
1.1735 0 Td
(min)Tj
1.8538 0 Td
(at)Tj
0.9525 0 Td
(5000)Tj
2.1203 0 Td
(rpm)Tj
-35.4611 -1.2982 Td
(at)Tj
0.9524 0 Td
(room)Tj
2.4208 0 Td
(temperature\))Tj
5.4708 0 Td
(were)Tj
2.1203 0 Td
(analysed)Tj
3.651 0 Td
(to)Tj
1.0262 0 Td
(assess)Tj
2.5228 0 Td
(total)Tj
2.0012 0 Td
(protein)Tj
3.152 0 Td
(content)Tj
3.2712 0 Td
([)Tj
0.83 0.64 0.02 0 k
(7)Tj
0 g
(])Tj
1.3776 0 Td
(and)Tj
1.7292 0 Td
(the)Tj
1.4683 0 Td
(molecular)Tj
4.2179 0 Td
(mass)Tj
-35.3817 -1.3039 Td
(distribution)Tj
4.9663 0 Td
(of)Tj
1.0148 0 Td
(proteins)Tj
3.5149 0 Td
(and)Tj
1.7291 0 Td
(peptides)Tj
3.5546 0 Td
(\(mono-dimension)Tj
7.3474 0 Td
(al)Tj
0.8956 0 Td
(SDS-PAGE,)Tj
5.0117 0 Td
(16%)Tj
1.9275 0 Td
(w/v)Tj
1.6894 0 Td
(acrylamide\))Tj
-31.6513 -1.2983 Td
([)Tj
0.83 0.64 0.02 0 k
(8)Tj
0 g
(].)Tj
1.5987 0 Td
(At)Tj
1.2019 0 Td
(least)Tj
1.9842 0 Td
(two)Tj
1.7122 0 Td
(biological)Tj
4.0875 0 Td
(replicates,)Tj
4.2122 0 Td
(with)Tj
1.9956 0 Td
(two)Tj
1.7121 0 Td
(technical)Tj
3.8098 0 Td
(replicates)Tj
3.9854 0 Td
(each,)Tj
2.2563 0 Td
(were)Tj
2.1203 0 Td
(performed)Tj
4.4674 0 Td
(for)Tj
1.389 0 Td
(all)Tj
-36.5326 -1.2983 Td
(the)Tj
1.4683 0 Td
(digestion)Tj
3.8835 0 Td
(conditions.)Tj
-4.1556 -1.2982 Td
(The)Tj
1.7858 0 Td
(three)Tj
2.2621 0 Td
(best)Tj
1.8084 0 Td
(digestion)Tj
3.8892 0 Td
(processes)Tj
3.9684 0 Td
(and)Tj
1.7292 0 Td
(the)Tj
1.4683 0 Td
(not)Tj
1.576 0 Td
(digested)Tj
3.5149 0 Td
(control)Tj
3.1181 0 Td
(at)Tj
0.9524 0 Td
(room)Tj
2.4265 0 Td
(temperature)Tj
5.1306 0 Td
(\(ND\))Tj
-34.8261 -1.304 Td
(were)Tj
2.1203 0 Td
(scaled-up)Tj
4.0308 0 Td
(in)Tj
1.0375 0 Td
(500)Tj
1.6441 0 Td
(mL)Tj
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(flasks)Tj
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(containing)Tj
4.5297 0 Td
(240)Tj
1.6441 0 Td
(mL)Tj
1.5817 0 Td
(of)Tj
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(final)Tj
2.0012 0 Td
(volume)Tj
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(reaction)Tj
3.498 0 Td
(\(six)Tj
1.6497 0 Td
(replicates)Tj
3.9855 0 Td
(for)Tj
-35.9203 -1.2982 Td
(each)Tj
2.0296 0 Td
(treatment\))Tj
4.473 0 Td
(using)Tj
2.3925 0 Td
(the)Tj
1.4683 0 Td
(following)Tj
3.9798 0 Td
(conditions:)Tj
4.6998 0 Td
(1\))Tj
1.0375 0 Td
(0.5)Tj
1.3946 0 Td
(U)Tj
0.9581 0 Td
(neu/g,)Tj
2.7382 0 Td
(pH)Tj
1.5023 0 Td
(7,)Tj
0.9241 0 Td
(50C,)Tj
2.4491 0 Td
(2h;)Tj
1.4514 0 Td
(2\))Tj
1.0374 0 Td
(0.5)Tj
1.3947 0 Td
(U)Tj
0.9581 0 Td
(alc/g,)Tj
-34.8885 -1.2983 Td
(pH)Tj
1.5023 0 Td
(7,)Tj
0.9241 0 Td
(60C,)Tj
2.4491 0 Td
(2h;)Tj
1.4514 0 Td
(3\))Tj
1.0318 0 Td
(0.5)Tj
1.4003 0 Td
(U)Tj
0.9524 0 Td
(pro/g,)Tj
2.6362 0 Td
(pH)Tj
1.5081 0 Td
(8,)Tj
0.9184 0 Td
(60C,)Tj
2.4548 0 Td
(2h.)Tj
1.4456 0 Td
(pH,)Tj
1.7348 0 Td
(dry)Tj
1.5704 0 Td
(weight,)Tj
3.1123 0 Td
(protein)Tj
3.1521 0 Td
(content)Tj
3.2712 0 Td
(and)Tj
1.7291 0 Td
(SDS-)Tj
-33.2444 -1.3039 Td
(PAGE)Tj
2.7496 0 Td
(molecular)Tj
4.2179 0 Td
(mass)Tj
2.194 0 Td
(distribution)Tj
4.9663 0 Td
(were)Tj
2.1259 0 Td
(analysed.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 351.4393 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Fractionation)Tj
5.9291 0 Td
(process)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 334.4881 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(The)Tj
1.7801 0 Td
(liquid)Tj
2.5625 0 Td
(supernatants)Tj
5.3461 0 Td
(\(about)Tj
2.8346 0 Td
(850)Tj
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(mL)Tj
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(each\))Tj
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(the)Tj
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(scaled-up)Tj
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(alc,)Tj
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(neu)Tj
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(and)Tj
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(pro)Tj
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(digestions)Tj
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(sub-fractionated)Tj
6.786 0 Td
(by)Tj
1.1793 0 Td
(cross-flow)Tj
4.3199 0 Td
(membrane)Tj
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(filtration)Tj
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(isolate)Tj
2.7609 0 Td
(peptide)Tj
3.1918 0 Td
(samples)Tj
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(having)Tj
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(weight)Tj
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(fractionation)Tj
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(was)Tj
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(a)Tj
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(Sartorius)Tj
3.8324 0 Td
(Slice)Tj
2.058 0 Td
(200)Tj
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(Benchtop)Tj
4.0421 0 Td
(system)Tj
2.9424 0 Td
(\(Sartorius-Stedim,)Tj
7.5571 0 Td
(Melsungen,)Tj
4.8869 0 Td
(Germany\))Tj
4.3256 0 Td
(using)Tj
2.3868 0 Td
(polyethersulfone)Tj
6.8938 0 Td
(\(PESU\))Tj
-33.0347 -1.3039 Td
(cassettes)Tj
3.5999 0 Td
(with)Tj
2.0013 0 Td
(a)Tj
0.6519 0 Td
(cut-off)Tj
2.9027 0 Td
(of)Tj
1.0148 0 Td
(0.2)Tj
/F16 1 Tf
1.4003 0 Td
[()]TJ
/F8 1 Tf
(m)Tj
1.542 0 Td
(\(0.02)Tj
2.2167 0 Td
(m)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 361.474 286.9228 cm
BT
/F8 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 365.0456 282.444 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(\),)Tj
0.7823 0 Td
(8)Tj
0.6973 0 Td
(kDa)Tj
1.8879 0 Td
(\(0.04)Tj
2.211 0 Td
(m)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 429.0519 286.9228 cm
BT
/F8 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 432.5669 282.444 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(\),)Tj
0.788 0 Td
(5)Tj
0.6973 0 Td
(kDa)Tj
1.8879 0 Td
(\(0.04)Tj
2.211 0 Td
(m)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 496.5732 286.9228 cm
BT
/F8 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 500.1448 282.444 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(\))Tj
0.5612 0 Td
(and)Tj
1.7291 0 Td
(1)Tj
0.6973 0 Td
(kDa)Tj
1.8879 0 Td
(\(0.04)Tj
-34.8885 -1.2982 Td
(m)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 208.1763 273.8834 cm
BT
/F8 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 211.748 269.4614 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(\))Tj
0.5612 0 Td
(molecular)Tj
4.2179 0 Td
(weight.)Tj
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1.7801 0 Td
(filtration)Tj
3.7474 0 Td
(was)Tj
1.7008 0 Td
(performed)Tj
4.4673 0 Td
(on)Tj
1.2756 0 Td
(ice)Tj
1.3323 0 Td
(using)Tj
2.3924 0 Td
(a)Tj
0.652 0 Td
(flow)Tj
1.9445 0 Td
(rate)Tj
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(of)Tj
1.0148 0 Td
(190)Tj
1.6441 0 Td
(mL/min)Tj
-32.7682 -1.2982 Td
(with)Tj
1.9955 0 Td
(a)Tj
0.652 0 Td
(maximum)Tj
4.3766 0 Td
(trans-membrane)Tj
6.9279 0 Td
(pressure)Tj
3.5773 0 Td
(of)Tj
1.0147 0 Td
(2)Tj
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(bar.)Tj
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1.7802 0 Td
(filtration)Tj
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(stopped)Tj
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(when)Tj
2.4038 0 Td
(the)Tj
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(retentate)Tj
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(volume)Tj
3.2031 0 Td
(reached)Tj
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(50)Tj
1.1679 0 Td
(mL.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 217.0204 cm
BT
/F7 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(In)Tj
1.0441 0 Td
(vitro)Tj
/F0 1 Tf
2.0882 0 Td
(biological)Tj
4.3181 0 Td
(activities)Tj
4.0063 0 Td
(assessment)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 200.0125 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Antioxidant)Tj
5.0626 0 Td
(capacity)Tj
3.4752 0 Td
(was)Tj
1.7008 0 Td
(measured)Tj
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(by)Tj
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(the)Tj
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(ABTS)Tj
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