Development and Validation of Analytical Method by HPLC-PDA and Seasonality from Gallic Acid, Catechin, and Epicatechin in Leaf Extracts from <i>Campomanesia adamantium</i>

Cabral, Raphael O.; Borges, Leonardo L.; Oliveira, Matheus G. de; Romano, Camila A.; Fiuza, Tatiana S.; Paula, José R. de

doi:10.21577/0103-5053.20230147

Acessibilidade / Reportar erro

Brasil

Journal of the Brazilian Chemical Society

Español English

Brasil

Español English

sumário « anterior atual seguinte »

Sumário

Article • J. Braz. Chem. Soc. 35 (3) • 2024 • https://doi.org/10.21577/0103-5053.20230147 copy

Development and Validation of Analytical Method by HPLC-PDA and Seasonality from Gallic Acid, Catechin, and Epicatechin in Leaf Extracts from Campomanesia adamantium

Authorship SCIMAGO INSTITUTIONS RANKINGS

Campomanesia adamantium is native of the Brazilian Cerrado, and its leaves are popularly used as anti-infammatory, antidiarrheal, and antiseptic in the urinary tract. This study aimed to develop and validate an analytical method by high-performance liquid chromatography for the simultaneous quantifcation and evaluation of the seasonal variability of gallic acid, catechin, and epicatechin, during 12 months, in C. adamantium leaf extracts. Chromatographic separations were performed with a mobile phase gradient of acetonitrile and 0.05% (v/v) trifuoroacetic acid solution, fow rate of 1.0 mL min⁻¹, detection at 210 nm, C18 column (150 × 4.6 mm, 5 μm) and column oven temperature of 35 °C. The analytical method developed was selective, linear, precise, accurate, robust, and without matrix effect. The parameters obtained in the present study meet the requirements established by national and international guidelines. The best time for leaves harvesting, with the highest levels reached, was in September (0.0626%, m/v) and August (0.044%, m/v) for gallic acid, in June (0.3953%, m/v) and July (0.3804%, m/v) for catechin, and from May (0.1622%, m/v) and June (0.1415%, m/v) for epicatechin, in the dry season. Therefore, this study contributes by providing parameters for quality control of the raw material C. adamantium.

Keywords:
Cerrado; chemical markers; gabiroba; quality control; seasonal variability; ultrasound-assisted extraction

Sample	System suitability parameters
Sample	k’	TF	Rs	N
GA standard (0.0327 mg mL⁻¹)	2.9 (0.34)	1.5 (0.95)	–^a a Resolution evaluates the ability to separate two consecutive peaks, but some samples that were analyzed did not present conditions to obtain this parameter. Values in parenthesis are the relative standard deviation percentage (RSD / %); k’: capacity factor; TF: tail factor; Rs: resolution; N: number of dishes; GA: gallic acid; CC: catechin; EC: epicatechin; US-FDA: United States Food and Drug Administration.	4,219.96 (1.39)
GA peak in the extract (17 mg mL⁻¹)	2.88 (0.21)	1.11 (1.41)	–^a a Resolution evaluates the ability to separate two consecutive peaks, but some samples that were analyzed did not present conditions to obtain this parameter. Values in parenthesis are the relative standard deviation percentage (RSD / %); k’: capacity factor; TF: tail factor; Rs: resolution; N: number of dishes; GA: gallic acid; CC: catechin; EC: epicatechin; US-FDA: United States Food and Drug Administration.	4,531.32 (0.63)
CC standard (0.0443 mg mL⁻¹)	7.29 (0.17)	1.13 (0.43)	–^a a Resolution evaluates the ability to separate two consecutive peaks, but some samples that were analyzed did not present conditions to obtain this parameter. Values in parenthesis are the relative standard deviation percentage (RSD / %); k’: capacity factor; TF: tail factor; Rs: resolution; N: number of dishes; GA: gallic acid; CC: catechin; EC: epicatechin; US-FDA: United States Food and Drug Administration.	16,437.27 (1.78)
CC peak in the extract (17 mg mL⁻¹)	7.25 (0.07)	1.06 (0.77)	17.82 (1.14)	18,887.7 (1.97)
EC standard (0.012 mg mL⁻¹)	8.94 (0.04)	1.2 (2.31)	–^a a Resolution evaluates the ability to separate two consecutive peaks, but some samples that were analyzed did not present conditions to obtain this parameter. Values in parenthesis are the relative standard deviation percentage (RSD / %); k’: capacity factor; TF: tail factor; Rs: resolution; N: number of dishes; GA: gallic acid; CC: catechin; EC: epicatechin; US-FDA: United States Food and Drug Administration.	14,917.74 (1.08)
EC peak in extract (17 mg mL⁻¹)	8.74 (0.10)	0.99 (1.77)	5.64 (0.92)	19,665.56 (1.77)
Literature specifcations (US-FDA)²⁴24 United States Food and Drug Administration (US-FDA); Reviewer Guidance, Validation of Chromatographic Methods; Center for Drug Evaluation and Research (CDER), Rockville, Maryland, United States of America, 1994. [Link ] accessed in August 2023 Link...	> 2	≤ 2	> 2	> 2000

Precision	Sample concentration / (mg mL⁻¹)	Area / (µV s⁻¹)	Marker concentration / (mg mL⁻¹)	Content / %	RSD / %
Gallic acid
Precision 1 day (analyst 1)-repeatability	13.6	369,864	0.0047	0.0342	1.15
	17	470,209	0.0059	0.0346
	20.4	575,439	0.0072	0.0351
Precision 2 day (analyst 2)	13.6	373,855	0.0047	0.0346	0.58
	17	469,261	0.0059	0.0345
	20.4	571,891	0.0071	0.0349
Intermediate precision				0.0346	0.89
Catechin
Precision 1 day (analyst 1)-repeatability	13.6	3,241,675	0.0358	0.2629	1.33
	17	4,158,007	0.0456	0.2684
	20.4	4,998,292	0.0547	0.2680
Precision 2 day (analyst 2)	13.6	3,217,704	0.0355	0.2610	1.85
	17	4,134,409	0.0454	0.2669
	20.4	5,046,474	0.0552	0.2705
Intermediate precision				0.2663	1.56
Epicatechin
Precision 1 day (analyst 1)-repeatability	13.6	887,435	0.0088	0.0649	1.39
	17	1,141,234	0.0113	0.0666
	20.4	1,370,794	0.0136	0.0666
Precision 2 day (analyst 2)	13.6	887,647	0.0088	0.0650	2.34
	17	1,128,032	0.0112	0.0659
	20.4	1,405,561	0.0139	0.0683
Intermediate precision				0.0662	1.88

Sample concentration / (mg mL⁻¹)	Marker area in the sample / (µV s⁻¹)	Marker area in swatch + marker pattern / (µV s⁻¹)	Recovered marker pattern concentration / (mg mL⁻¹)	Recovery / %	RSD / %
Gallic acid (theoretical concentration of standard GA added: 0.0025 mg mL⁻¹)
13.6	367,722	566,520	0.0026	101.79	1.14
17	477,382	675,633	0.0026	101.53
20.4	573,835	770,133	0.0026	100.59
Catechin (theoretical concentration of CC standard added: 0.0141 mg mL⁻¹)
13.6	3,232,637	4,448,419	0.0139	99.24	1.23
17	4,192,389	5,435,546	0.0142	101.33
20.4	5,012,151	6,229,671	0.0140	99.37
Epicatechin (theoretical concentration of EC pattern added: 0.0068 mg mL⁻¹)
13.6	882,714	1,558,156	0.0067	99.14	0.99
17	1,112,191	1,791,895	0.0068	99.75
20.4	1,329,356	2,004,407	0.0067	99.08

Parameter	Area of GA / (µV s⁻¹)	Area of CC / (µV s⁻¹)	Area of EC / (µV s⁻¹)	Content of GA / %	Content of CC / %	Content of EC / %
Original method	469,749	4,186,501	1,160,040	0.0345	0.2702	0.0677
Column oven temperature / °C
34	468,180 (1.04)	4,115,517 (1.16)	1,167,272 (0.93)	0.0344 (1.01)	0.2657 (1.14)	0.0682 (0.92)
36	469,464 (0.72)	4,078,673 (1.57)	1,138,796 (1.17)	0.0345 (0.70)	0.2633 (1.54)	0.0665 (1.16)
Mobile phase pH
2.5	485,307 (1.90)	4,137,986 (0.98)	1,232,848 (3.41)	0.0356 (1.85)	0.2671 (0.96)	0.0719 (3.38)
2.7	487,159 (2.11)	4,154,149 (1.01)	1,235,828 (3.49)	0.0358 (2.06)	0.2661 (0.99)	0.0721 (3.46)
Flow rate / (mL min⁻¹)
0.9	481,252 (1.52)	4,187,375 (0.65)	1,224,298 (3.01)	0.0354 (1.48)	0.2702 (0.64)	0.0715 (2.99)
1.1	479,405 (1.36)	4,101,216 (1.28)	1,223,608 (3.12)	0.0352 (1.32)	0.2648 (1.26)	0.0715 (3.09)

Date	Total rainfall / mm	Relative humidity / %	Average temperature / °C
Date	Total rainfall / mm	Relative humidity / %	Maximum	Minimum
02/20/2015	155.1	68.9	33.2	21
03/31/2015	156.2	67.8	32.3	21.2
04/30/2015	1.3	50.3	33.4	20.2
05/31/2015	70.7	66	29.7	18.4
06/30/2015	0	56.3	30.2	17
07/31/2015	2.7	50.9	31.4	16.7
08/31/2015	3.6	38.4	33.3	17.5
09/30/2015	30.4	42.5	30.4	20.3
10/31/2015	18.2	43.5	36.7	22.1
11/30/2015	354.8	63.9	33.9	21
12/31/2015	207.7	66	33	21.2
01/31/2016	484.8	80.2	29.8	21

Date	Average GA content / %	RSD / %	Average CC content / %	RSD / %	Average EC content / %	RSD / %
02/20/2015	0.0393	1.95	0.2663	0.62	0.0390	0.65
03/31/2015	0.0414	1.47	0.2410	0.45	0.0430	0.5
04/30/2015	0.0423	2.29	0.2540	0.67	0.0482	0.5
05/31/2015	0.0401	0.57	0.3783	0.98	0.1622	0.95
06/30/2015	0.0357	2.73	0.3953	1.09	0.1415	1
07/31/2015	0.0344	1.53	0.3804	0.21	0.1231	0.44
08/31/2015	0.0440	0.79	0.3416	1.18	0.0734	1.32
09/30/2015	0.0626	0.19	0.3275	0.33	0.0035	2.68
10/31/2015	0.0313	0.34	0.1599	0.9	–	-
11/30/2015	0.0336	1.28	0.1839	0.31	0.0067	0.63
12/31/2015	0.0350	1.77	0.2041	0.87	0.0164	0.57
01/31/2016	0.0355	2.45	0.2252	0.65	0.0244	1.87

	Eingenvalues functions	Canonical R	Wilk’s Lambda	p-level^a
F1^b	5.17	0.91	0.13	0.0015
	Standardized coeffcients for Canonical variables
Gallic acid	−1.22
Catechin	1.49
Eigenvalues	5.17
Cumulative proportion	0.951

	Total well-classifcation / %	Cluster I	Cluster II	Cluster III
		p = 0.5	p = 0.5
Cluster I	100	5	0	0
Cluster II	50	2	2	0
Cluster III	100	0	0	3
Total	83.3	7	2	3

Sociedade Brasileira de Química Instituto de Química - UNICAMP, Caixa Postal 6154, 13083-970 Campinas SP - Brazil, Tel./FAX.: +55 19 3521-3151 - São Paulo - SP - Brazil
E-mail: office@jbcs.sbq.org.br

Acompanhe os números deste periódico no seu leitor de RSS

[1] * e-mail: jose_realino@ufg.br