Development of an analytical method for determination of polyphenols and total tannins from leaves of <i>Syzygium cumini</i> L. Skeels

GOMES, CAMILA L.; SILVA, CAIO C.A.R.; MELO, CAMILA G. DE; FERREIRA, MAGDA R.A.; SOARES, LUIZ A.L.; DA SILVA, ROSALI M.F.; ROLIM, LARISSA A.; ROLIM NETO, PEDRO J.

doi:10.1590/0001-3765202120190373

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CELLULAR AND MOLECULAR BIOLOGY • An. Acad. Bras. Ciênc. 93 (2) • 2021 • https://doi.org/10.1590/0001-3765202120190373 linkcopy

Development of an analytical method for determination of polyphenols and total tannins from leaves of Syzygium cumini L. Skeels

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Syzygium cumini L. Skeels belongs to Myrtaceae family. This species has been recognized by its antidiabetic, anti-inflammatory, and antimicrobial activities. Despite ever-increasing scientific interest for this species there is no pharmacopeia method for characterization and standardization of S. cumini yet. So, toward this aim, the objective of this work was to develop an efficient analytical methodology able to determine polyphenols and tannins content from leaves hydroethanolic extract of S. cumini using Folin-Ciocalteu method by ultraviolet absorption spectrophotometry (UV-Vis). The analytical methodology was developed for the first time in the literature for leaves of this specie shown to be fast and low-cost with results expressed through tannic acid equivalent (TAE). Moreover, the methodology presented selectivity with maximum absorption at 706 nm wavelength, linearity with R²>0.99; limit of detection 0.275 µg TAE mL^-1 and 0.102 µg TAE mL^-1; limit of quantification 1.046 µg TAE mL^-1 and 0.912 µg TAE mL^-1 for total polyphenols and total tannins, respectively. Furthermore, the methodology was accurate with recover value greater than 98%, as well as exact, reproductive, and robust with coefficient of variation values less than 15% for both compounds. All the results are found within the fixed limits according to RDC 166/2017.

Key words
Polyphenols; Syzygium cumini; Tannins; UV-Vis

Parameters	TT (mg TAE g^-1 ext) ± SD	CV (%)
Casein 100 mg	2.378 ± 0.013	0.55
Casein 250 mg	2.403 ± 0.009	0.37
Casein 500 mg	2.622 ± 0.063	2.29
Casein 750 mg	2.701 ± 0.008	0.31
Casein 1000 mg	2.912 ± 0.006	0.19

	Merits evaluation	Standard Solution	Extractive Solution
TP	LOD	0.083 µg mL^-1	0.275 µg TAE mL^-1
	LOQ	0.314 µg mL^-1	1.046 µg TAE mL^-1
	Cochran`s Test	0.674 (p-value)	1 (p-value)
	Durbin-Watson Test	0.298 (p-value)	0.963 (p-value)
TT	LOD	0.100 µg mL^-1	0.102 µg TAE mL^-1
	LOQ	0.894 µg mL^-1	0.912 µg TAE mL^-1
	Cochran`s Test	1 (p-value)	0.709 (p-value)
	Durbin-Watson Test	0.963 (p-value)	0.961 (p-value)

Parameters	TP level µg TAE mL^-1 ± SD	TT level µg TAE mL^-1 ± SD	F. calculated TP/TT	F. tabulated TP/TT
Sodium Carbonate 10%	6.194 ± 0.009	5.777 ± 0.118	1.189/ 1.997	5.318/ 5.591
Sodium Carbonate 15%	6.266 ± 0.146	5.652 ± 0.140	1.189/ 1.997	5.318/ 5.591
Shimadzu® Equipment	6.365 ± 0.472	6.334 ± 0.212	3.572/1.847	7.709/ 7.709
Micronal® Equipment	6.732 ± 0.162	6.615 ± 0.289	3.572/1.847	7.709/ 7.709

Compund	Parameters	Level µg TAE mL^-1 ± SD (CV%) (1)	Level µg TAE mL^-1 ± SD (CV%) (2)	F _tabulated	F _calculated
TP	Days 1 e 2	7.169 ± 0.708 (9.87)	7.211 ± 0.119 (1.65)	5.317	1.222
	Analysts 1 e 2	6.233 ± 0.072 (1.16)	6.151 ± 0.103 (1.68)	3.466	0.020
TT	Days 1 e 2	6.280 ± 0.113 (1.80)	6.261 ± 0.829 (13.24)	5.317	0.001
	Analysts 1 e 2	5.695 ± 0.091 (1.59)	5.858 ± 0.080 (1.37)	7.708	5.467

Compound	Extractive Solution + TA	Theoretical Value	Real Value ± SD	Recovery
TP	4.80 µg TAE mL^-1 + 1 µg TA mL^-1	5.80 µg TAE mL^-1	5.77 µg TAE mL^-1 ±0.002	99.48%
	6.00 µg TAE mL^-1 + 1 µg TA mL^-1	7.00 µg TAE mL^-1	6.97 µg TAE mL^-1 ±0.005	99.57%
	7.20 µg TAE mL^-1 + 1 µg TA mL^-1	8.20 µg TAE mL^-1	8.05 µg TAE mL^-1 ±0.002	98.17%
TT	4.20 µg TAE mL^-1 + 1 µg TA mL^-1	5.20 µg TAE mL^-1	5.20 µg TAE mL^-1 ±0.017	100,00%
	5.14 µg TAE mL^-1 + 1 µg TA mL^-1	6.14 µg TAE mL^-1	6.15 µg TAE mL^-1 ±0.019	100.16%
	6.05 µg TAE mL^-1 + 1 µg TA mL^-1	7.05 µg TAE mL^-1	7.05 µg TAE mL^-1 ±0.019	100.00%

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[tfnI] Work data; CV = Coefficient of Variation; SD = Standard Deviation; TAE = Tannic Acid; Equivalent; TT = Total Tannins.