Microwave-assisted extraction, HPLC analysis, and inhibitory effects on carbonic anhydrase I, II, VA, and VII isoforms of 14 blueberry Italian cultivars

Figures & data

Figure 1. Chemical structures of the reported analytes.

Table 1. Multi-component pattern for hydroalcoholic extracts of fourteen Italian cultivars expressed as μg/mg of dry extract.

Cultivar	Delphinidin-3-O- glucoside	Malvidin-3-O- galactoside	Cyanidin-3-O- galactoside	Delphinidin-3-O- galactoside	Cyanidin-3-O- rutinoside	Malvidin-3-O- glucoside	Chlorogenic acid	Total
Prima Donna	0.72	3.58	1.24	10.34	4.03	1.10	1.26	22.27
Chandler	2.84	6.91	1.69	9.68	20.02	nd	2.33	43.46
Emerald	2.26	3.57	1.30	12.58	4.86	2.82	1.94	29.34
Misty	1.27	3.01	nd	3.77	1.63	2.57	10.05	22.30
Jewels	2.55	2.35	1.23	8.83	2.98	3.77	2.18	23.89
Bluetta	2.32	3.57	nd	nd	1.45	2.52	2.82	12.68
Blue Crop	2.77	4.42	2.82	13.55	5.74	5.39	5.46	40.14
Gulf Coast	4.59	8.99	2.02	16.52	5.96	1.86	4.56	44.52
Spartan	2.83	3.50	0.35	13.31	nd	2.63	3.67	26.27
Palmetto	1.02	2.06	nd	5.34	1.67	nd	0.90	11.00
Snow Chaser	0.97	2.78	nd	8.86	2.89	4.31	4.57	24.39
Rebel	1.43	1.53	0.43	5.72	nd	1.61	1.04	11.75
Camelia	nd	3.43	1.05	3.98	1.96	1.58	3.10	15.10
Spring Wind	9.77	4.27	nd	23.87	nd	7.61	11.81	57.34
Total	35.34	53.97	12.13	136.35	53.19	37.77	55.69
Mean value	2.52	3.85	0.87	9.74	3.80	2.70	3.98

nd, not detected.

Table 2. Multi-component pattern for organic extracts of fourteen Italian cultivars expressed as μg/mg of dry extract.

Cultivar	Delphinidin-3-O- glucoside	Malvidin-3-O- galactoside	Cyanidin-3-O- galactoside	Delphinidin-3-O- galactoside	Cyanidin-3-O- rutinoside	Malvidin-3-O- glucoside	Chlorogenic acid	Total
Prima Donna	nd	nd	nd	nd	nd	nd	nd
Chandler	nd	nd	nd	nd	nd	nd	nd
Emerald	nd	nd	nd	nd	nd	nd	0.56	0.56
Misty	nd	nd	nd	nd	nd	nd	1.79	1.79
Jewels	nd	nd	nd	nd	nd	nd	nd
Bluetta	nd	nd	nd	nd	nd	nd	nd
Blue Crop	nd	nd	nd	nd	nd	nd	nd
Gulf Coast	nd	13.41	nd	nd	nd	18.19	15.51	47.11
Spartan	nd	nd	nd	nd	nd	nd	6.88	6.88
Palmetto	nd	nd	nd	nd	nd	nd	4.07	4.07
Snow Chaser	nd	nd	nd	nd	nd	nd	2.83	2.83
Rebel	nd	nd	nd	nd	nd	nd	nd
Camelia	nd	nd	nd	nd	nd	nd	nd
Spring Wind	nd	nd	nd	nd	nd	nd	nd
Total	nd	13.41	nd	nd	nd	18.19	31.64
Mean value	0	0.96	0	0	0	1.30	2.26

nd, not detected.

Table 3. Inhibition data of human CA isoforms hCA I, II, VA, and VII for reported Blueberry cultivar extracts by a stopped flow CO₂ hydrase assay. O, organic extract; A, hydroalcoholic extract.

	IC50 (μg/mL)*
Extract	hCA I	hCA II	hCA VA	hCA VII
Prima Donna A	>1000	380	0.007	1.59
Prima Donna O	134.7	>1000	0.35	0.17
Blue Crop A	>1000	>1000	2.38	2.73
Blue Crop O	308	>1000	0.07	0.24
Emerald A	>1000	>1000	>1000	>1000
Emerald O	>1000	>1000	>1000	>1000
Jewels A	>1000	>1000	>1000	>1000
Jewels O	>1000	>1000	>1000	>1000
Mysty A	>1000	>1000	>1000	>1000
Mysty O	>1000	>1000	>1000	>1000
Chandler A	>1000	>1000	>1000	>1000
Chandler O	>1000	>1000	0.087	>1000
Spring Wind A	>1000	>1000	>1000	>1000
Spring Wind O	>1000	>1000	>1000	>1000
Bluetta A	>1000	>1000	>1000	>1000
Bluetta O	0.072	0.082	0.045	>1000
Rebel A	>1000	0.73	0.73	0.059
Rebel O	>1000	>1000	>1000	>1000
Palmetto A	>1000	>1000	>1000	>1000
Palmetto O	>1000	>1000	>1000	>1000
Gulf Coast A	>1000	0.63	0.47	0.52
Gulf Coast O	>1000	0.027	0.077	>1000
Snow Chaser A	>1000	0.17	0.0009	0.25
Snow Chaser O	>1000	>1000	>1000	>1000
Spartan A	>1000	>1000	>1000	>1000
Spartan O	>1000	0.0063	>1000	>1000
Camelia A	>1000	0.22	0.008	0.082
Camelia O	>1000	0.070	>1000	>1000

*From three different assays, errors ± 5–10% of the reported value.

Table 4. Inhibition data of human CA isoforms hCA I, II, VA, and VII of the two representative anthocyanins, chlorogenic acid and the standard sulfonamide inhibitor acetazolamide (AAZ) by a stopped flow CO₂ hydrase assay.

	Ki (μM)*
Compound	hCA I	hCA II	hCA VA	hCA VII
Delphinidin-3-O-galactoside	>100	>100	18.7	>100
Cyanidin-3-O-rutinoside	>100	>100	609	>100
Chlorogenic acid	25.0	30.1	0.05	>100
Acetazolamide (AAZ)	250	12	25	6.0

*From three different assays, errors ± 5–10% of the reported value.