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Comparative Study on Physicochemical, Antioxidant and Antimicrobial Properties of Propolis Collected from Different Regions of Bulgaria

Yulian Tumbarski
Yulian Tumbarski
, 
Mina Todorova
Mina Todorova
, 
Mariyana Topuzova
Mariyana Topuzova
, 
Gabriela Gineva
Gabriela Gineva
, 
Velichka Yanakieva
Velichka Yanakieva
, 
Ivan Ivanov
Ivan Ivanov
 et 
Nadezhda Petkova
Nadezhda Petkova
   | 30 juin 2023
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Article Category: Original Article
Publié en ligne: 30 juin 2023
Pages: 37 - 56
Reçu: 22 déc. 2022
Accepté: 02 mai 2023
DOI: https://doi.org/10.2478/jas-2023-0004
Mots clés
© 2023 Yulian Tumbarski et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

TPC, TFC and antioxidant activity of the methanolic propolis extracts (20 mg/ml)

Sample # Polyphenols, mg GAE/g Flavonoids, mg QE/g Antioxidant activity
DPPH, mM TE/g FRAP, mM TE/g
1 172.89 ± 0.55 90.10 ± 0.39 982.01 ± 4.98 584.33 ± 1.62
2 212.77 ± 0.80 129.24 ± 4.99 1080.27 ± 5.48 738.66 ± 1.78
3 536.33 ± 1.00 182.59 ± 4.02 1598.66 ± 3.67 966.77 ± 2.44
4 72.30 ± 0.53 59.07 ± 0.38 18.56 ± 1.70 136.26 ± 1.26
5 111.56 ± 0.38 63.95 ± 1.63 372.21 ± 5.27 368.55 ± 2.58
6 183.66 ± 0.36 113.53 ± 1.13 1064.84 ± 7.28 637.44 ± 1.32
7 164.98 ± 0.60 90.58 ± 1.20 746.78 ± 2.72 566.27 ± 2.08
8 121.77 ± 1.36 73.64 ± 0.18 761.13 ± 4.62 440.20 ± 3.62
9 159.15 ± 0.41 85.24 ± 1.82 692.42 ± 5.60 548.80 ± 2.88
10 160.30 ± 0.75 83.36 ± 1.53 885.39 ± 5.09 535.41 ± 3.27
11 110.83 ± 0.91 121.17 ± 0.57 514.78 ± 3.89 954.01 ± 6.28
12 216.91 ± 0.62 132.88 ± 4.77 998.45 ± 3.03 1208.81 ± 5.25
13 191.39 ± 0.55 107.92 ± 0.39 946.54 ± 5.17 1188.24 ± 5.33
14 133.79 ± 0.54 73.94 ± 1.53 534.07 ± 2.47 715.80 ± 4.47
15 129.46 ± 0.53 71.18 ± 1.34 662.57 ± 5.38 839.27 ± 2.42
16 181.81 ± 0.56 90.84 ± 1.39 949.44 ± 4.88 1067.59 ± 1.04
17 136.24 ± 0.98 68.06 ± 1.36 575.78 ± 5.31 792.54 ± 2.27
18 102.14 ± 0.86 49.24 ± 1.81 512.78 ± 6.34 637.39 ± 4.58
19 167.08 ± 0.94 82.11 ± 1.79 816.89 ± 3.04 1046.72 ± 1.66
20 127.53 ± 0.34 69.59 ± 1.29 624.89 ± 6.00 773.27 ± 2.99
21 161.80 ± 0.79 92.06 ± 1.95 769.91 ± 4.32 1008.77 ± 5.01
22 160.23 ± 0.79 74.42 ± 1.00 616.31 ± 6.77 889.43 ± 5.78
23 117.84 ± 1.02 60.48 ± 1.08 572.03 ± 4.56 507.99 ± 1.81
24 133.63 ± 0.41 76.41 ± 0.44 587.83 ± 4.27 564.24 ± 7.34
25 134.74 ± 0.21 77.44 ± 0.88 568.52 ± 7.02 580.02 ± 4.83
26 161.10 ± 1.23 92.70 ± 1.76 725.14 ± 2.30 726.39 ± 3.79
27 110.50 ± 0.81 48.08 ± 0.21 341.12 ± 1.62 439.23 ± 5.39
28 157.02 ± 0.62 94.37 ± 2.40 569.24 ± 3.38 629.84 ± 4.53
29 144.70 ± 0.62 74.17 ± 1.52 521.70 ± 3.02 616.84 ± 7.29
30 143.96 ± 0.80 68.23 ± 1.90 477.54 ± 3.48 574.52 ± 2.47
31 151.05 ± 1.05 83.11 ± 0.88 600.98 ± 7.05 689.24 ± 2.48
32 137.32 ± 0.82 65.73 ± 0.43 479.85 ± 6.94 600.13 ± 2.11
33 158.38 ± 0.42 70.96 ± 1.99 580.17 ± 3.21 647.47 ± 2.02
34 146.42 ± 0.21 76.27 ± 2.47 608.38 ± 4.32 616.81 ± 4.45
35 137.84 ± 0.42 72.93 ± 2.55 543.90 ± 2.98 639.19 ± 4.04
36 141.08 ± 0.87 54.90 ± 0.69 531.72 ± 4.91 636.80 ± 4.61
37 145.39 ± 1.24 65.93 ± 2.84 451.22 ± 3.19 590.35 ± 2.61
38 175.85 ± 0.62 123.80 ± 3.50 880.94 ± 6.44 925.23 ± 3.61
39 179.40 ± 0.38 86.82 ± 0.61 749.55 ± 2.52 835.86 ± 5.34
40 213.16 ± 0.34 97.85 ± 2.54 816.25 ± 5.11 811.95 ± 2.24
41 63.14 ± 0.85 29.22 ± 1.80 21.25 ± 1.16 98.91 ± 0.57
42 93.69 ± 0.51 60.94 ± 1.26 76.79 ± 1.15 82.28 ± 1.69
43 133.38 ± 1.03 57.93 ± 0.18 680.20 ± 2.27 404.47 ± 4.59
44 145.05 ± 0.57 84.30 ± 3.01 606.80 ± 3.37 514.01 ± 4.89
45 157.82 ± 0.66 91.60 ± 2.09 623.41 ± 4.92 551.75 ± 5.86
46 160.13 ± 0.56 99.31 ± 3.14 673.64 ± 2.52 510.15 ± 6.52
47 135.15 ± 0.88 77.85 ± 0.56 586.01 ± 5.89 470.76 ± 4.08
48 119.58 ± 0.73 57.16 ± 0.38 450.65 ± 2.39 407.17 ± 1.15
49 165.38 ± 0.27 104.41 ± 1.28 701.95 ± 4.97 536.42 ± 4.80
50 120.14 ± 0.61 54.92 ± 1.34 319.64 ± 2.27 357.21 ± 1.79
51 152.67 ± 0.60 78.13 ± 0.84 543.50 ± 4.36 510.88 ± 3.54
52 177.78 ± 1.85 124.61 ± 0.95 1013.64 ± 3.07 1009.48 ± 2.00
53 158.35 ± 1.78 94.02 ± 0.48 898.27 ± 2.98 981.19 ± 3.01
54 154.72 ± 2.10 79.96 ± 0.88 707.57 ± 5.29 644.32 ± 2.19
55 125.49 ± 0.47 72.20 ± 0.99 691.62 ± 2.09 719.70 ± 2.03
56 135.91 ± 0.44 68.33 ± 1.15 833.28 ± 3.60 513.52 ± 2.38
57 162.14 ± 0.80 84.99 ± 1.45 958.49 ± 2.46 639.48 ± 5.09
58 148.78 ± 1.76 94.15 ± 1.39 982.84 ± 1.94 643.46 ± 2.50
59 148.04 ± 0.11 90.10 ± 0.47 1145.43 ± 6.85 695.37 ± 3.62
60 125.72 ± 2.84 62.30 ± 1.61 902.38 ± 4.28 768.80 ± 4.55
61 89.25 ± 0.96 54.22 ± 1.13 360.69 ± 1.92 574.67 ± 1.38
62 127.01 ± 3.33 74.98 ± 1.58 694.29 ± 6.99 376.74 ± 2.77
63 190.83 ± 3.46 152.69 ± 1.18 1026.52 ± 2.15 594.17 ± 3.62
64 131.54 ± 0.70 64.26 ± 0.50 668.90 ± 2.82 552.82 ± 2.66
65 154.95 ± 3.57 89.97 ± 1.10 440.13 ± 1.09 460.33 ± 2.96
66 194.26 ± 3.67 98.48 ± 0.72 538.23 ± 5.63 538.87 ± 4.15
67 197.35 ± 1.50 102.36 ± 0.49 955.39 ± 2.99 758.80 ± 6.43
68 170.02 ± 0.70 85.07 ± 0.74 891.30 ± 1.56 763.06 ± 5.26
69 145.10 ± 0.11 76.52 ± 0.67 534.79 ± 1.57 482.80 ± 2.55
70 124.69 ± 0.34 78.59 ± 0.47 672.02 ± 2.77 461.11 ± 3.75
71 159.38 ± 1.60 93.35 ± 1.44 753.68 ± 4.20 531.95 ± 4.60
72 152.72 ± 0.96 84.53 ± 0.90 517.65 ± 3.89 640.56 ± 1.00
73 163.78 ± 1.43 96.37 ± 0.69 1165.56 ± 3.74 918.94 ± 7.80
74 188.50 ± 1.75 100.04 ± 0.65 1192.44 ± 7.60 863.00 ± 2.63
75 234.18 ± 3.30 129.85 ± 0.70 902.11 ± 1.94 842.92 ± 2.33
76 234.29 ± 1.46 151.53 ± 1.42 1459.09 ± 3.63 1012.26 ± 3.61
77 737.27 ± 1.08 234.17 ± 1.14 1464.75 ± 3.46 916.64 ± 5.66
78 103.20 ± 2.32 50.12 ± 1.40 397.38 ± 2.01 522.24 ± 5.90
79 206.25 ± 0.86 110.31 ± 0.46 1149.19 ± 2.45 884.08 ± 3.32
80 158.50 ± 3.71 83.74 ± 0.85 832.65 ± 4.99 959.66 ± 4.88

Origin of the propolis samples

Sample # Town/village District GPS Coordinates Climatic zone
1 Stamboliyski Plovdiv 42°08′N 24°36′E TrC
2 Silistra Silistra 44°07′N 27°17′E TC
3 Simitli Blagoevgrad 41°54′N 23°08′E CM
4 Burgas Burgas 42°30′N 27°28′E BS
5 Ruse Ruse 43°50′N 25°57′E TC
6 Kozarevets Stara Zagora 42°13′N 25°34′E TrC
7 Valchedram Montana 43°45′N 23°26′E TC
8 Targovishte Targovishte 43°15′N 26°34′E TC
9 Matenitsa Plovdiv 42°19′N 24°22′E TrC
10 Brezovo Plovdiv 42°20′N 25°08′E TrC
11 Krichim Plovdiv 42°03′N 24°28′E TrC
12 Gorna Malina Sofia 42°41′N 23°42′E TC
13 Shumen Shumen 43°16′N 26°55′E TC
14 Varna Varna 43°13′N 27°55′E BS
15 Krasnovo Plovdiv 42°27′N 24°32′E TrC
16 Asenovgrad Plovdiv 42°01′N 24°52′E TrC
17 Batak (Tsarnat) Pazardzhik 41°57′N 24°13′E M
18 Batak (St. Troitsa) Pazardzhik 41°57′N 24°13′E M
19 Shabla Dobrich 43°49′N 27°09′E BS
20 Lenovo Plovdiv 41°57′N 25°04′E TrC
21 Hisarya Plovdiv 42°30′N 24°42′E TrC
22 Karnobat Burgas 42°39′N 26°59′E TrC
23 Dimitrovgrad Haskovo 42°03′N 25°36′E TrC
24 Glavan Stara Zagora 42°03′N 26°07′E TrC
25 Tsarimir Plovdiv 42°19′N 24°40′E TrC
26 Dren Pernik 42°25′N 23°09′E TC
27 Negovantsi Pernik 42°27′N 22°57′E TC
28 Divlya Pernik 42°32′N 22°40′E TrC
29 Vrabcha Pernik 42°51′N 22°44′E TC
30 Montana Montana 43°25′N 23°13′E TC
31 Bezdenitsa Montana 43°55′N 23°20′E TC
32 Barkach Pleven 43°18′N 24°25′E TC
33 Pernik Pernik 42°36′N 23°02′E TC
34 Osenets Razgrad 43°33′N 26°22′E TC
35 Popina Silistra 44°07′N 26°57′E TC
36 Loznitsa Dobrich 43°58′N 27°55′E TC
37 Debrene Dobrich 43°23′N 27°51′E TC
38 Boychinovtsi Montana 43°27′N 23°22′E TC
39 Bankya Sofia 42°42′N 23°08′E TC
40 Vladimir Pernik 42°26‘N 23°05‘E TC
41 Pernik (Tsarkva) Pernik 42°36′N 23°02′E TC
42 Petrich Blagoevgrad 41°24′N 23°13′E CM
43 Yakoruda Blagoevgrad 42°01′N 23°41′E M
44 Pernik (Draganovets) Pernik 42°36′N 23°02′E TC
45 Brezovo 2 Plovdiv 42°20′N 25°08′E TrC
46 Chirpan Stara Zagora 42°12′N 25°20′E TrC
47 Razgrad Razgrad 43°32′N 26°31′E TC
48 Belovitsa Plovdiv 42°25′N 24°31′E TrC
49 Lenovo 2 Plovdiv 41°57′N 25°04′E TrC
50 Lenovo 3 Plovdiv 41°57′N 25°04′E TrC
51 Yundola Pazardzhik 42°03′N 23°52′E M
52 Dyulevo Burgas 42°23′N 27°09′E BS
53 Zagortsi Burgas 42°24′N 27°04′E BS
54 Krusheto V. Tarnovo 43°13′N 25°40′E TC
55 Dzhurovo Sofia 42°56′N 24°03′E M
56 Dolna Dikanya Pernik 42°25′N 23°07′E TC
57 Bornarevo Pernik 42°32′N 22°55′E TC
58 Cherkovo Burgas 42°29′N 27°02′E TrC
59 Nisovo Ruse 43°39′N 26°04′E TC
60 Belene Pleven 43°39′N 25°07′E TC
61 Aleksandrovo Shumen 42°59′N 26°58′E TC
62 Elena V. Tarnovo 42°56′N 25°53′E TC
63 Cherven breg Kyustendil 42°18′N 23°10′E TrC
64 Gorsko Novo selo V. Tarnovo 43°05′N 25°56′E TC
65 Borovitsa Kardzhali 41°66′N 25°22′E CM
66 Simeonovgrad Haskovo 42°02′N 25°50′E TrC
67 Gamzovo Vidin 44°05′N 22°45′E TC
68 Medeshevtsi Vidin 43°53′N 22°38′E TC
69 Voynitsa Vidin 43°56′N 22°41′E TC
70 Kotel Sliven 42°53′N 26°27′E TrC
71 Dolna Kremena Vratsa 43°10′N 23°44′E TC
72 Smolyan Smolyan 41°35′N 24°42′E CM
73 Bulgarsko Slivovo V. Tarnovo 43°33′N 25°18′E TC
74 Parsha Gabrovo 42°57′N 25°29′E TC
75 Ritya Gabrovo 42°59′N 25°25′E TC
76 Kozi rog Gabrovo 42°57′N 25°16′E TC
77 Burya Gabrovo 43°02′N 25°19′E TC
78 Yambol Yambol 42°29′N 26°30′E TrC
79 Malinovo Lovech 42°90′N 24°90′E TC
80 Starosel Plovdiv 42°29′N 24°34′E TrC

Colour index of the raw propolis samples

Sample # Colour Colour index
L a b C-ab hᵒ
1 Light brown 55.81 ± 5.01 5.02 ± 0.26 8.88 ± 0.11 10.26 ± 2.49 59.45 ± 7.0
2 Light brown 54.44 ± 1.19 5.99 ± 1.02 8.98 ± 1.13 10.80 ± 1.35 56.42 ± 1.8
3 Brown 50.50 ± 5.63 6.80 ± 2.67 6.91 ± 0.63 9.74 ± 4.97 43.24 ± 6.7
4 Dark brown 43.42 ± 2.88 1.54 ± 0.35 1.07 ± 0.35 1.89 ± 0.25 35.40 ± 8.9
5 Dark brown 49.48 ± 0.78 4.72 ± 0.24 3.61 ± 0.16 5.94 ± 0.25 37.46 ± 1.9
6 Dark brown 45.54 ± 1.25 2.75 ± 0.50 3.38 ± 0.91 4.37 ± 0.40 51.17 ± 4.0
7 Dark green 47.93 ± 2.12 2.65 ± 0.87 2.44 ± 0.37 3.61 ± 1.39 41.73 ± 4.1
8 Dark green 47.89 ± 2.30 3.00 ± 1.07 2.58 ± 0.70 4.04 ± 0.74 41.90 ± 4.0
9 Brown 50.23 ± 4.12 4.85 ± 0.54 7.33 ± 0.91 8.82 ± 1.05 56.12 ± 6.7
10 Green 52.68 ± 2.51 1.97 ± 0.47 1.43 ± 0.18 2.44 ± 0.44 36.52 ± 5.3
11 Brown 49.04 ± 1.28 3.67 ± 0.50 3.48 ± 0.47 5.10 ± 0.72 43.08 ± 8.7
12 Light brown 58.22 ± 1.71 6.71 ± 0.73 19.20 ± 3.03 20.35 ± 3.30 70.53 ± 2.8
13 Light brown 54.81 ± 0.84 8.28 ± 0.84 18.86 ± 0.38 20.62 ± 4.63 65.82 ± 3.3
14 Dark brown 42.53 ± 2.15 2.84 ± 0.23 3.67 ± 0.93 4.67 ± 0.74 51.59 ± 7.2
15 Dark green 40.52 ± 0.63 5.01 ± 0.17 8.53 ± 0.55 9.89 ± 0.08 59.58 ± 1.3
16 Dark brown 43.45 ± 2.33 11.07 ± 0.41 6.75 ± 0.19 12.97 ± 0.43 31.35 ± 2.0
17 Brown 50.00 ± 2.27 6.55 ± 1.37 9.13 ± 0.89 11.36 ± 2.65 53.50 ± 1.4
18 Dark brown 38.92 ± 2.22 2.80 ± 0.19 2.74 ± 0.92 3.93 ± 0.22 44.20 ± 5.3
19 Brown 49.74 ± 1.59 5.65 ± 0.30 10.78 ± 1.23 12.21 ± 2.68 61.54 ± 5.9
20 Dark brown 46.75 ± 0.66 7.53 ± 0.73 13.52 ± 1.74 15.48 ± 0.83 60.92 ± 1.4
21 Brown 51.85 ± 0.51 6.19 ± 0.14 10.67 ± 0.42 12.34 ± 0.10 59.86 ± 1.0
22 Green 51.94 ± 0.13 5.34 ± 0.48 12.78 ± 2.52 13.85 ± 1.00 67.34 ± 0.4
23 Dark brown 42.41 ± 2.58 3.98 ± 0.33 5.10 ± 0.72 6.49 ± 0.72 51.71 ± 5.7
24 Dark brown 39.84 ± 2.33 4.47 ± 0.42 3.70 ± 0.97 5.89 ± 0.41 39.06 ± 2.3
25 Light brown 52.49 ± 0.12 6.80 ± 0.36 11.38 ± 0.56 13.27 ± 1.65 58.92 ± 2.9
26 Dark brown 47.51 ± 1.03 5.98 ± 0.54 6.12 ± 1.91 8.57 ± 0.18 45.66 ± 4.4
27 Light brown 54.67 ± 0.65 7.85 ± 0.58 19.58 ± 1.00 21.10 ± 2.56 68.07 ± 1.0
28 Light brown 53.74 ± 0.42 7.03 ± 0.67 26.43 ± 1.21 27.36 ± 0.63 75.09 ± 1.6
29 Brown 50.11 ± 3.31 3.55 ± 0.14 3.08 ± 0.51 4.73 ± 0.70 40.18 ± 8.1
30 Light brown 54.82 ± 2.07 4.41 ± 0.12 7.62 ± 0.78 8.80 ± 0.55 59.86 ± 1.1
31 Dark brown 45.66 ± 4.31 6.31 ± 1.78 7.36 ± 0.65 9.71 ± 3.32 48.42 ± 4.8
32 Dark brown 45.90 ± 1.14 4.64 ± 0.21 7.92 ± 1.18 9.18 ± 0.96 59.49 ± 2.3
33 Brown 50.86 ± 4.13 6.60 ± 0.84 19.00 ± 1.62 20.12 ± 1.39 70.88 ± 1.4
34 Dark brown 36.65 ± 3.55 6.36 ± 0.15 11.61 ± 2.16 13.25 ± 0.38 61.25 ± 1.6
35 Dark brown 44.85 ± 1.45 6.69 ± 0.22 11.45 ± 0.41 13.27 ± 0.79 59.66 ± 0.9
36 Light brown 54.01 ± 0.77 4.71 ± 1.11 8.46 ± 0.88 9.70 ± 1.07 61.19 ± 4.3
37 Dark brown 45.05 ± 2.35 5.11 ± 0.36 8.82 ± 0.20 10.19 ± 1.19 59.78 ± 1.9
38 Brown 51.89 ± 1.99 3.86 ± 0.72 6.47 ± 1.23 7.53 ± 1.75 58.95 ± 1.9
39 Light brown 55.57 ± 1.02 5.55 ± 0.62 16.78 ± 1.52 17.68 ± 2.16 71.61 ± 2.1
40 Brown 50.91 ± 2.03 6.47 ± 0.92 19.18 ± 0.39 20.25 ± 0.66 71.40 ± 2.2
41 Dark brown 42.34 ± 1.14 3.92 ± 0.33 10.73 ± 0.23 11.42 ± 0.85 69.88 ± 2.0
42 Dark brown 45.48 ± 2.68 1.99 ± 0.77 9.29 ± 0.48 2.01 ± 0.78 8.17 ± 0.5
43 Dark brown 45.96 ± 1.63 8.17 ± 0.41 16.23 ± 0.59 18.17 ± 1.27 63.24 ± 0.8
44 Dark brown 48.31 ± 0.97 8.54 ± 3.49 17.69 ± 0.43 19.77 ± 2.66 64.78 ± 7.7
45 Dark brown 45.51 ± 5.21 8.26 ± 0.38 19.84 ± 0.34 21.49 ± 0.34 67.39 ± 1.1
46 Brown 51.65 ± 2.20 7.47 ± 0.21 17.97 ± 0.43 19.46 ± 0.22 67.42 ± 0.6
47 Dark brown 36.37 ± 0.70 4.89 ± 0.33 9.04 ± 0.81 10.28 ± 0.56 61.58 ± 0.8
48 Light brown 59.75 ± 4.77 8.95 ± 0.78 25.06 ± 0.48 26.62 ± 0.81 70.37 ± 1.2
49 Green 49.88 ± 0.37 5.23 ± 0.61 18.19 ± 0.11 18.93 ± 0.41 73.97 ± 1.9
50 Light brown 57.09 ± 0.45 7.38 ± 0.20 21.91 ± 1.13 23.12 ± 0.37 71.39 ± 0.3
51 Dark brown 32.27 ± 1.19 4.61 ± 0.28 7.45 ± 0.63 8.76 ± 0.47 58.22 ± 1.4
52 Brown 51.08 ± 0.87 7.75 ± 0.24 14.80 ± 1.35 16.71 ± 0.82 62.36 ± 0.6
53 Light brown 52.27 ± 0.20 8.86 ± 0.22 23.28 ± 0.16 24.91 ± 0.39 69.15 ± 0.8
54 Brown 50.37 ± 3.19 5.66 ± 1.37 17.21 ± 0.55 18.14 ± 0.70 71.85 ± 4.0
55 Light brown 51.51 ± 2.63 5.24 ± 1.09 8.73 ± 2.39 10.19 ± 2.58 58.79 ± 2.7
56 Light brown 53.77 ± 2.43 5.86 ± 2.16 20.22 ± 3.94 21.07 ± 4.38 74.16 ± 2.7
57 Brown 48.19 ± 0.55 7.99 ± 2.12 17.57 ± 1.81 19.33 ± 2.49 65.84 ± 3.8
58 Brown 52.23 ± 2.31 5.85 ± 2.15 22.27 ± 4.19 23.04 ± 4.56 75.62 ± 2.9
59 Dark brown 45.15 ± 6.27 2.10 ± 0.60 1.94 ± 0.31 2.86 ± 0.64 43.27 ± 4.0
60 Green 50.94 ± 3.92 4.71 ± 0.74 9.21 ± 1.59 10.35 ± 1.71 62.83 ± 2.2
61 Green 53.08 ± 6.74 4.96 ± 1.89 14.55 ± 2.79 15.46 ± 5.76 69.90 ± 8.3
62 Dark brown 39.47 ± 0.80 2.48 ± 0.25 3.41 ± 0.49 4.23 ± 0.33 53.76 ± 5.9
63 Light brown 51.50 ± 3.72 11.30 ± 2.73 25.99 ± 1.96 28.42 ± 1.93 66.55 ± 5.7
64 Light brown 51.87 ± 3.44 7.94 ± 1.45 20.84 ± 1.67 22.31 ± 2.05 69.26 ± 2.1
65 Light brown 56.38 ± 0.64 5.30 ± 1.71 21.35 ± 4.66 22.01 ± 4.93 76.24 ± 1.4
66 Light brown 59.04 ± 1.62 5.99 ± 0.61 19.20 ± 2.67 20.12 ± 2.72 72.63 ± 0.8
67 Green 51.60 ± 0.90 5.88 ± 0.72 14.02 ± 2.76 15.25 ± 2.42 66.73 ± 5.5
68 Green 53.48 ± 1.49 6.17 ± 0.53 21.50 ± 0.39 22.37 ± 0.40 74.00 ± 1.3
69 Green 52.38 ± 1.32 7.04 ± 0.22 22.33 ± 1.39 23.42 ± 1.39 72.48 ± 0.6
70 Dark brown 40.23 ± 0.45 4.66 ± 0.17 9.67 ± 0.24 10.73 ± 0.27 64.27 ± 0.5
71 Brown 47.02 ± 1.20 9.45 ± 0.34 16.31 ± 0.24 18.85 ± 0.37 59.92 ± 0.6
72 Brown 45.62 ± 1.21 6.81 ± 0.80 10.21 ± 0.97 12.27 ± 1.16 56.33 ± 2.3
73 Dark green 35.00 ± 4.78 4.46 ± 0.84 12.76 ± 2.18 13.52 ± 2.33 70.77 ± 0.6
74 Green 48.95 ± 2.70 5.11 ± 0.38 18.45 ± 1.60 19.15 ± 1.62 74.50 ± 0.8
75 Green 46.15 ± 0.95 6.76 ± 0.51 16.41 ± 0.21 17.75 ± 0.37 67.63 ± 1.3
76 Dark brown 41.99 ± 5.17 7.49 ± 1.08 16.02 ± 2.23 17.72 ± 5.15 63.96 ± 5.1
77 Green 55.15 ± 0.25 6.71 ± 2.49 11.58 ± 1.64 13.43 ± 2.67 60.64 ± 5.5
78 Dark green 43.56 ± 3.52 5.55 ± 1.26 9.43 ± 1.06 10.95 ± 1.54 59.80 ± 3.1
79 Green 51.85 ± 1.86 4.09 ± 0.46 6.27 ± 0.38 7.49 ± 0.57 56.93 ± 1.4
80 Brown 45.66 ± 1.00 4.39 ± 0.33 11.79 ± 0.95 12.59 ± 0.83 69.47 ± 2.6

Antimicrobial activity of the methanolic propolis extracts (20 mg/ml)

Test microorganism Inhibition zones (IZ), mm*
Minimum Maximum Average MeOH (control)
B. subtilis ATCC 6633 15 25 18.91 ± 1.63 0
B. amyloliquefaciens 4BCL-YT 8 19 15.43 ± 1.56 0
S. aureus ATCC 25923 13 25 19.95 ± 3.05 0
L. monocytogenes NBIMCC 8632 12 30 19.65 ± 3.51 0
L. innocua ATCC 33090 12 24 18.15 ± 2.71 0
E. faecalis ATCC 19433 12 19 14.74 ± 1.83 0
E. faecium ATCC 19434 11 18 14.88 ± 1.35 0
M. luteus 2YC-YT 15 28 19.99 ± 2.56 0
S. enteritidis ATCC 13076 11 18 14.09 ± 1.15 0
Klebsiella sp. (clinical isolate) 8 11 8.94 ± 0.94 0
E. coli ATCC 25922 12 20 16.12 ± 1.71 0
P. vulgaris ATCC 6380 8 12 8.56 ± 1.06 0
P. aeruginosa ATCC 9027 8 22 13.41 ± 4.96 0
C. albicans NBIMCC 74 8 15 12.10 ± 1.37 0
S. cerevisiae ATCC 9763 9 17 12.85 ± 1.53 0
A. niger ATCC 1015 10 16 12.56 ± 1.11 0
A. flavus (plant isolate) 10 20 14.30 ± 2.73 0
Penicillium sp. (plant isolate) 9 18 13.11 ± 1.32 0
Rhizopus sp. (plant isolate) 10 30 17.72 ± 5.68 0
F. moniliforme ATCC 38932 8 17 12.27 ± 2.17 0

Correlation between the TPC, TFC and antioxidant activity of the propolis samples

Methods Correlation, r2
TPC / TFC 0.85
TPC / DPPH 0.68
TPC / FRAP 0.44
TFC / DPPH 0.78
TFC / FRAP 0.55
DPPH / FRAP 0.68

Interrelationship between the colour, colour components (L, a, b, C-ab, and hᵒ), TPC, TFC, and antioxidant activity of the propolis samples

Parameter Groups
Colour LB B DB G DG
TPC 157.46 ± 32.57 179.87 ± 91.20 136.86* ± 34.40 214.96 ± 168.8 136.64* ± 27.07
TFC 87.85 ± 27.88 96.81 ± 32.30 76.50 ± 22.68 101.42 ± 46.93 76.38 ± 18.20
DPPH 698.2 ± 239.0 789.0 ± 304.6 605.9 ± 331.1 879.4 ± 302.7 746.4 ± 276.0
FRAP 665.4 ± 235.5 771.9 ± 190.7 555.6 ± 236.4 734.6 ± 159.2 657.2 ± 209.4
L ≤40 40–45 45–50 50–55 ≥55
TPC 137.26 ± 19.92 138.34 ± 50.35 150.16 ± 28.20 169.07* ± 27.78 236.93 ± 204.0
TFC 75.61 ± 14.84 76.17 ± 31.51 85.81 ± 19.02 89.78 ± 29.13 105.56 ± 54.48
DPPH 671.1 ± 202.8 638.3 ± 438.8 679.7 ± 236.5 773.0 ± 258.1 742.6 ± 370.7
FRAP 585.0 ± 150.3 623.9 ± 319.7 626.6 ± 215.0 716.4 ± 185.5 663.5 ± 331.4
a ≤2 2–4 4–6 6–8 ≥8
TPC 110.32 ± 48.53 136.63 ± 33.22 154.49 ± 23.05 199.14* ± 85.05 159.73* ± 24.95
TFC 67.79 ± 13.52 81.24 ± 28.56 81.51 ± 17.65 98.16* ± 42.83 92.20 ± 28.23
DPPH 327.0 ± 484.1 664.1 ± 295.5 714.8 ± 239.0 774.7** ± 338.2 770.2 ± 194.9
FRAP 251.0 ± 247.4 597.6 ± 231.8 665.2 ± 169.7 730.3** ± 211.1 693.2 ± 300.2
b ≤5 5–10 10–15 15–20 ≥20
TPC 133.38 ± 28.87 166.48 ± 85.71 183.22 ± 151.20 174.15* ± 34.50 148.38 ± 21.36
TFC 78.90 ± 19.97 82.78 ± 35.11 86.14 ± 35.95 96.25* ± 26.76 84.68 ± 27.05
DPPH 637.9 ± 308.0 706.0 ± 360.0 709.4 ± 371.1 795.1** ± 274.3 692.0 ± 244.9
FRAP 553.5 ± 195.6 655.7 ± 206.0 653.9 ± 206.9 709.4** ± 253.4 580.3 ± 175.9
C-ab ≤5 5–10 10–15 15–20 ≥20
TPC 132.46 ± 33.64 175.89 ± 102.6 173.94 ± 100.62 166.36* ± 35.43 157.83* ± 28.90
TFC 76.69 ± 17.64 89.20 ± 33.48 85.24 ± 32.15 95.46* ± 22.75 87.74 ± 26.40
DPPH 632.9 ± 356.1 692.5 ± 313.2 725.4 ± 272.7 827.6** ± 216.9 691.9 ± 234.9
FRAP 494.3 ± 216.1 686.3 ± 187.4 721.0 ± 235.0 686.6** ± 175.2 653.8 ± 249.4
hᵒ, ≤40 40–50 50–60 60–70 ≥70
TPC 128.12 ± 42.64 182.31 ± 134.3 153.91 ± 28.86 175.57 ± 115.49 157.86 ± 29.96
TFC 72.06 ± 12.98 95.50 ± 37.82 84.09 ± 21.36 90.73 ± 40.12 85.69 ± 19.94
DPPH 470.7 ± 392.9 799.1 ± 356.1 704.2 ± 223.1 754.5 ± 298.8 712.7 ± 262.3
FRAP 467.3 ± 361.8 693.1 ± 174.7 645.6 ± 143.9 708.4 ± 250.8 657.4 ± 211.7

pH and moisture content the propolis samples

Parameter Minimum Maximum Average*
pH 4.82 ± 0.02 5.87 ± 0.01 5.32 ± 0.18
Moisture, % 0.98 ± 0.25 2.97 ± 0.35 1.76 ± 0.41
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