Optimization of the Extraction Process of Bioactive Compounds from <i>Zingiber officinale</i> Roscoe, Evaluation of Acetylcholinesterase Enzyme Inhibition and Cytotoxic Activity of the Free and Encapsulated Extract

Nardino, Danielli A.; Aranha, Ana C. R.; Tonin, Lilian T. D.; Defendi, Rafael O.; Ishikawa, Sabrina; Bressiani, Patricia A.; Santana, Ana B. S.; Dusman, Elisângela; Yonekawa, Murilo K. A.; Jaques, Jeandre A. S.; Santos, Edson A.; Tonin, Angélica P. P.; Meurer, Eduardo C.; Sipoli, Caroline C.; Suzuki, Rúbia M.

doi:10.21577/0103-5053.20230186

Danielli A. Nardino

original draft

data analysis

figures (preparation and editing) formal analysis

methodology

writing (review and editing)

discussed the results and contributed to the final manuscript

Departamento de Química, Universidade Estadual de Maringá (UEM), 87020-900 Maringá-PR, Brazil

Ana C. R. Aranha

figures (editing, formal analysis)

writing (review and editing)

discussed the results and contributed to the final manuscript

Departamento de Engenharia Química, Universidade Estadual de Maringá (UEM), 87020-900 Maringá-PR, Brazil

https://orcid.org/0000-0002-9576-4868

Lilian T. D. Tonin

formal analysis

review and editing

discussed the results and contributed to the final manuscript

Programa de Pós-Graduação em Engenharia Química (PPGEQ-AP), Universidade Tecnológica Federal do Paraná (UTFPR), 86812-460 Apucarana-PR, Brazil

Rafael O. Defendi

formal analysis

review and editing

discussed the results and contributed to the final manuscript

Programa de Pós-Graduação em Engenharia Química (PPGEQ-AP), Universidade Tecnológica Federal do Paraná (UTFPR), 86812-460 Apucarana-PR, Brazil

Sabrina Ishikawa

investigation cytotoxic analysis

discussed the results and contributed to the final manuscript

Departamento de Química e Biologia, Universidade Tecnológica Federal do Paraná (UTFPR), 85602-863 Francisco Beltrão-PR, Brazil

Patricia A. Bressiani

investigation cytotoxic analysis

discussed the results and contributed to the final manuscript

Departamento de Química e Biologia, Universidade Tecnológica Federal do Paraná (UTFPR), 85602-863 Francisco Beltrão-PR, Brazil

Ana B. S. Santana

investigation cytotoxic analysis

discussed the results and contributed to the final manuscript

Departamento de Química e Biologia, Universidade Tecnológica Federal do Paraná (UTFPR), 85602-863 Francisco Beltrão-PR, Brazil

Elisângela Dusman

investigation

supervision cytotoxic analysis

formal analysis

review

discussed the results and contributed to the final manuscript

Departamento de Química e Biologia, Universidade Tecnológica Federal do Paraná (UTFPR), 85602-863 Francisco Beltrão-PR, Brazil

Murilo K. A. Yonekawa

investigation AChE enzyme inhibition

discussed the results and contributed to the final manuscript

Setor de Bioquímica, Instituto de Biociências, Universidade Federal do Mato Grosso do Sul (UFMS), 79070-900 Campo Grande-MS, Brazil

Jeandre A. S. Jaques

resources

discussed the results and contributed to the final manuscript

Setor de Bioquímica, Instituto de Biociências, Universidade Federal do Mato Grosso do Sul (UFMS), 79070-900 Campo Grande-MS, Brazil

Edson A. Santos

investigation supervision AChE enzyme inhibition

formal analysis

review

discussed the results and contributed to the final manuscript

Setor de Bioquímica, Instituto de Biociências, Universidade Federal do Mato Grosso do Sul (UFMS), 79070-900 Campo Grande-MS, Brazil

Angélica P. P. Tonin

investigation mass spectrometry data processing

discussed the results and contributed to the final manuscript

Laboratório Fenn de Espectrometria de Massas, Departamento de Química, Universidade Federal do Paraná (UFPR), 86900-000 Jandaia do Sul-PR, Brazil

Eduardo C. Meurer ^** e-mail: eduardo.meurer@ufpr.br

supervision mass spectrometry

discussed the results and contributed to the final manuscript

Laboratório Fenn de Espectrometria de Massas, Departamento de Química, Universidade Federal do Paraná (UFPR), 86900-000 Jandaia do Sul-PR, Brazil

Programa de Pós-Graduação em Biotecnologia Ambiental (PEB), Universidade Estadual de Maringá (UEM), 87020-900 Maringá-PR, Brazil

https://orcid.org/0000-0003-4835-7773

Caroline C. Sipoli

review and supervision

discussed the results and contributed to the final manuscript

Programa de Pós-Graduação em Engenharia Química (PPGEQ-AP), Universidade Tecnológica Federal do Paraná (UTFPR), 86812-460 Apucarana-PR, Brazil

Rúbia M. Suzuki

review and supervision

discussed the results and contributed to the final manuscript

Programa de Pós-Graduação em Engenharia Química (PPGEQ-AP), Universidade Tecnológica Federal do Paraná (UTFPR), 86812-460 Apucarana-PR, Brazil

* e-mail: eduardo.meurer@ufpr.br

Editor handled this article: Andrea R. Chaves (Associate)

Independent variables	Coded and real levels of independent variables
Independent variables	–1.41	–1	0	+1	+ 1.41
Temperature / °C	20	25	40	55	60
time / h	0.5	4	12	20	24

Component	Condition 1	Condition 2
Desolvation gas temperature / °C	200	250
Source gas temperature / °C	110	110
Ionization mode	MS^-	MS^-
Capillary voltage / kV	2.0	2.5
Cone voltage / V	20.0	40.0
Collision energy / V	15.0	30.0
Collision gas pressure / mbar	3.5 × 10^-3	3.5 × 10^-3

Run	Temperature / °C	time / h	ABTS / %	DPPH / %	CA / %
1	25	4	21.05 ± 0.21^g	82.20 ± 0.13^f,^g	92.63 ± 0.15^b,^c
2	55	20	27.29 ± 0.15^b	82.52 ± 0.08 ^e,^f	99.90 ± 0.02^a
3	40	0.5	23.36 ± 0.17^d,^e	86.34 ± 0.15^b	88.61 ± 0.15f
4	25	20	27.60 ± 0.27^b	88.49 ± 0.13^a	90.35 ± 0.06^e
5	40	12	22.39 ± 0.22^f	82.46 ± 0.17^e,^f	91.60 ± 0.09^d
6	60	12	29.87 ± 0.21^a	83.58 ± 0.14^d	99.78 ± 0.06^a
7	40	12	22.58 ± 0.09^e,^f	82.78 ± 0.15^e	91.28 ± 0.18^d
8	55	4	23.52 ± 0.07^d	81.77 ± 0.05^g	92.28 ± 0.13^c
9	20	12	24.70 ± 0.11^c	85.17 ± 0.05^c	87.15 ± 0.22^g
10	40	12	23.14 ± 0.04^d,^e,^f	82.49 ± 0.08^e,^f	91.27 ± 0.21^d
11	40	24	23.74 ± 0.17^d	85.66 ± 0.11^c	93.22 ± 0.11^b

	Source of variation	Df	SS	MS	F value	Pr (> F)
ABTS	T	1	60.03	67.03	37.71	7.15 × 10⁸ ^a
	T	1	88.61	88.61	49.85	1.97 × 10⁹ ^a
	T²	1	158.92	158.92	89.40	1.71 × 10^-13 ^a
	t²	1	2.67	2.67	1.50	0.22
	T × t	1	11.55	11.55	6.50	0.01^b
	residuals	60	106.66	1.78
DPPH	T	1	56.08	56.08	33.68	2.61 × 10^{-7 a}
	T	1	27.75	27.75	16.67	1.34 × 10^{-4 a}
	T²	1	0.11	0.11	0.06	0.80
	t²	1	39.08	39.08	23.47	9.31 × 10^{-6 a}
	T × t	1	46.06	46.06	27.66	2.03 × 10^{-6 a}
	residuals	60	99.90	1.67
CA	T	1	548.66	548.66	345.80	2.20 × 10^-16 ^a
	T	1	105.75	105.75	66.65	2.57 × 10^{-11 a}
	T²	1	72.18	72.18	45.49	6.80 × 10^-9 ^a
	t²	1	0.97	0.97	0.61	0.44
	T × t	1	146.83	146.83	92.54	9.08 × 10^{-14 a}
	residuals	60	95.20	1.59

	ABTS		DPPH		CA
	PC 1	PC 2	PC 1	PC 2	PC 1	PC 2
T	0.9997	0.0213	–0.4407	0.8976	0.9237	–0.3832
t	–0.0213	0.9997	0.8976	0.4407	0.3832	0.9237
Variance / %	64.03	35.97	74.92	25.08	93.77	6.23

Analyte	[M – H]^-	MRM transition (m/z)	Condition 1 (S/N:RMS)	Condition 2 (S/N:RMS)	Reference
Malic acid	133	133 > 115	98	–	33
Galic acid	169	169 > 125	65	–	33
Ferrulic acid	193	193 > 134	–	97	35
6-Gingerol	293	293 > 99; 193	820; 289	997	32
8-Shogaol	303	303 > 167	125	193	37
8-Gingerol	321	321 > 127	160	–	32
10-Shogaol	331	331 > 195	205	–	37
10-Gingerol	349	349 > 155; 193	323; 110	–	32
Quercetina-3-O-hexosideo	463	463 > 179	63	–	34

	Ginger extract / %
	0	10	20	30
D_h / nm	184.73 ± 0.52^a	174.45 ± 0.70^a	168.04 ± 5.02^a	168.06 ± 5.14^a
Polydispersity / %	26.17 ± 1.14^a	25.33 ± 0.50^a	25.93 ± 0.29^a	26.23 ± 0.74^a
ζ / mV	–34.80 ± 0.40^a	31.80 ± 0.93^a	30.80 ± 1.12^a	31.47 ± 1.13^a

Group	VC / %
	Human kidney tumor cells (786-0)			Human liver tumor cells (HUH7.5)			Monkey kidney normal cell
	24 h	48 h	72 h	24 h	48 h	72 h	24 h	48 h	72 h
CO^-	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00
CO⁺	65.24	45.09	20.74	39.76	12.94	27.31	6.32	35.52	29.63
5 µg mL^-1	115.77	122.77	100.00	98.06	112.23	97.33	106.31	94.13	117.98
10 µg mL^-1	99.89	121.97	122.00	101.67	115.91	105.51	95.49	106.83	101.39
50 µg mL^-1	121.57	145.20	94.39	105.55	106.04	87.84	98.72	102.05	101.33
100 µg mL^-1	98.93	136.07	111.21	85.15	93.06	89.63	101.83	106.63	90.84
200 µg mL^-1	136.48	127.86	89.75	98.33	77.05	85.49	108.06	84.90	93.62
300 µg mL^-1	132.08	138.15	71.67	95.63	58.07	97.81	115.57	74.68	94.56
400 µg mL^-1	113.52	108.55	56.06	75.92	56.42	66.37	78.76	64,.94	92.67
500 µg mL^-1	106.44	93.87	48.70	62.04	49.01	37.03	115.87	131.63	78.80
1000 µg mL^-1	52.25	38.03	25.38	99.93	27.01	31.28	121.55	110.86	63.76

Group	VC human kidney tumor (786-0)
	0%			10%			20%			30%
	24 h	48 h	72 h	24 h	48 h	72 h	24 h	48 h	72 h	24 h	48 h	72 h
CO^-	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00
CO⁺	104.79	118.24	98.42	93.29	81.75	103.05	65.82	67.96	67.01	55.35	75.93	80.92
5 µg mL^-1	100.96	159.04	106.98	86.84	100.16	72.58	80.03	63.74	65.65	84.38	93.05	67.84
10 µg mL^-1	95.40	91.84	85.51	89.56	90.37	65.61	83.89	97.55	65.60	101.03	67.95	72.51
50 µg mL^-1	106.13	119.20	114.36	135.33	89.74	85.24	86.20	83.69	71.74	119.75	77.84	72.95
100 µg mL^-1	80.46	103.36	111.46	96.22	86.53	90.05	75.92	85.55	70.22	76.29	65.60	110.67
200 µg mL^-1	137.36	108.48	66.53	106.35	81.75	97.16	99.37	95.91	56.85	85.25	69.56	81.87
300 µg mL^-1	97.32	142.40	82.21	127.16	79.56	84.97	102.79	96.87	56.97	91.67	71.10	80.85
400 µg mL^-1	95.79	93.92	94.99	87.52	84.10	94.43	88.74	86.64	59.11	69.63	74.88	72.51
500 µg mL^-1	162.64	97.60	95.52	94.30	84.34	66.22	91.70	66.36	74.45	96.35	56.76	83.92
1000 µg mL^-1	139.85	146.72	76.42	83.21	60.22	71.29	102.55	59.51	51.72	56.15	60.22	148.25

Group	VC human hepatoma
	0%			10%			20%			30%
	24 h	48 h	72 h	24 h	48 h	72 h	24 h	48 h	72 h	24 h	48 h	72 h
CO^-	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00
CO⁺	58.72	59.40	31.43	37.71	40.63	51.36	55.41	49.42	27.05	104.33	41.16	36.67
5 µg mL^-1	85.61	108.80	106.25	128.24	71.67	84.63	133.02	98.14	107.06	102.11	106.48	163.26
10 µg mL^-1	71.85	97.07	90.92	106.57	134.58	79.02	111.82	96.52	123.37	116.86	122.03	170.03
50 µg mL^-1	83.99	141.40	110.57	108.91	83.23	100.81	106.93	111.95	133.75	130.09	87.58	176.91
100 µg mL^-1	100.90	154.00	119.54	97.94	95.00	120.16	115.54	129.00	135.77	144.73	91.23	159.40
200 µg mL^-1	89.57	117.21	99.61	86.87	102.40	94.76	96.88	129.47	129.18	128.57	81.33	166.62
300 µg mL^-1	87.41	139.65	117.49	88.56	122.92	108.86	117.99	125.64	122.48	107.26	77.64	120.28
400 µg mL^-1	89.30	130.77	106.03	99.16	135.42	132.64	102.20	106.03	126.81	108.08	75.23	185.08
500 µg mL^-1	107.55	117.05	110.74	92.12	99.48	110.31	97.30	110.44	132.98	137.94	97.56	199.21
1000 µg mL^-1	75.36	97.46	81.52	77.20	91.77	69.17	83.61	88.05	129.77	125.64	95.73	157.83

Group	VC normal monkey kidney
	0%			10%		20%			30%
	24 h	48 h	72 h	24 h	48 h	72 h	24 h	48 h	72 h	24 h	48 h	72 h
CO^–	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00
CO⁺	10.05	6.08	43.83	5.25	4.47	32.00	16.70	42.16	41.65	6.76	28.09	34.48
5 µg mL^-1	82.46	119.69	103.74	113.47	139.22	90.72	103.85	107.74	99.11	111.51	111.12	124.00
10 µg mL^-1	100.99	141.17	112.67	77.49	128.75	108.35	94.67	105.48	90.13	108.73	104.20	107.63
50 µg mL^-1	106.99	142.21	138.76	130.81	131.91	110.50	137.06	135.56	123.39	80.70	113.90	107.18
100 µg mL^-1	141.38	161.90	94.68	97.23	136.42	111.36	132.15	145.31	99.85	97.24	111.07	97.87
200 µg mL^-1	281.22	446.27	316.02	120.98	133.27	99.75	118.35	141.54	115.00	97.49	90.24	121.15
300 µg mL^-1	242.56	341.86	314.63	114.46	151.65	78.26	117.29	94.59	89.24	115.82	96.42	103.30
400 µg mL^-1	276.21	456.75	403.29	112.72	136.26	107.25	115.39	109.94	117.97	92.09	75.77	138.75
500 µg mL^-1	189.50	439.30	317.92	98.97	108.63	62.59	95.56	114.22	64.51	102.49	124.80	73.42
1000 µg mL^-1	334.53	452.75	379.90	71.21	40.02	59.77	65.90	52.70	84.43	98.13	51.70	42.63

[1] * e-mail: eduardo.meurer@ufpr.br

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Optimization of the Extraction Process of Bioactive Compounds from Zingiber officinale Roscoe, Evaluation of Acetylcholinesterase Enzyme Inhibition and Cytotoxic Activity of the Free and Encapsulated Extract