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

Tumbarski, Yulian; Todorova, Mina; Topuzova, Mariyana; Gineva, Gabriela; Yanakieva, Velichka; Ivanov, Ivan; Petkova, Nadezhda

doi:10.2478/jas-2023-0004

Figures & Tables

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

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

Figures & Tables

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

Origin of the propolis samples

Colour index of the raw propolis samples

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

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

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

pH and moisture content the propolis samples

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