Effect of extraction time on antioxidants and bioactive volatile components of green tea (Camellia sinensis), using GC/MS

Figures & data

Figure 1. (a)–(c) Showing GC–MS chromatograph of leaf-grade tea type at different extraction times

Figure 2. (a)–(c) Showing GC–MS chromatograph of sanding-grade tea type at different extraction times

Table 1. (a)–(c) Showing the identification and relative percentage of volatile compounds in leaf-grade type of green tea at 50°C in 20-, 40-, and 120-min time of extraction

Peak	R Time	Area %	Name of compound
(a) Leaf grade, 20 min
4	12.01	8.19	Heptadecanoic acid and methyl ester
5	13.558	0.8	Cyclononasiloxane and octadecamethyl
6	14.589	24.43	Palmitic Acid and hexadecanoic acid
7	15.959	0.67	1H-purine-6-amine
8	16.219	3.85	Phthalic Acid
9	17.331	2.24	Stearic acid, methyl ester, octadecanoic acid, and methyl ester
10	17.493	36.94	9-octadecenoic acid, methyl ester, and oleic acid
12	19.568	1.95	12-octadecadienoic acid, methyl ester, and linoleic acid
13	20.512	1.64	Tetradecanoic acid and myristic acid
16	28.268	7.42	Caffein
(b) Leaf grade, 40 min
4	12.02	5.77	Methyl heptadecanoate
6	14.607	18.31	Palmitic acid, methyl ester, hexadecanoic acid, and methyl ester
7	15.89	0.34	Phenol, 2-methoxy-4-(1-propenyl)
8	16.012	0.35	1H-purine-6-amine
9	16.228	3.73	Phthalic Acid
10	17.349	0.57	Octadecanoic acid, methyl ester, stearic acid, and methyl ester
11	17.506	31.24	9-octadecenoic acid, methyl ester, oleic acid, and methyl ester
12	17.917	5.57	Linoleic acid and 9,12-octadecadienoic acid
14	19.584	3.36	13,16-octadecadienoic acid and methyl ester
15	20.247	3.23	Tetradecanoic acid and myristic acid
18	28.287	18.08	Caffein
(c) Leaf grade, 120 min
3	9.75	0.16	Methyl hexadecanoate
5	12.019	6.41	Methyl heptadecanoate
7	14.599	19.22	Methyl hexadecanoate, palmitic acid, and methyl ester
8	15.888	0.38	Phenol, 2-methoxy-4-(1-propenyl)-
9	15.996	0.73	1H-purine-6-amine,
10	16.225	3.45	Phthalic acid
11	17.342	1.61	Methyl octadecanoate, stearic acid, and methyl ester
12	17.502	33.09	9-octadecenoic acid, methyl ester, oleic acid, and methyl ester
13	17.916	1.43	9,12-octadecadienoic acid, methyl ester, and linoleic acid
15	19.584	1.49	9,12-octadecadienoic acid and methyl ester
16	20.496	1.42	Tetradecanoic acid
18	21.629	0.43	12,15-octadecadiynoic acid and methyl ester
20	28.278	17.8	Caffein

Table 2. (a)–(c) Showing the identification and relative percentage of volatile compounds in sanding type of green tea at 40°C in 20-, 40-, and 120-min time of extraction

Peak	R Time	Area %	Compound name
(a) Sanding, 20 min
4	11.477	0.25	Carbamic acid, phenyl ester, and phenyl carbamate
5	12.022	4.26	Methyl tetradecanoate and tetradecanoic acid
6	13.332	0.37	Phenol and 2-methoxy-4-2-propenyl
8	14.608	13.98	Hexadecanoic acid and methyl ester
9	15.894	0.76	Phenol, 2-methoxy-4-(1-propenyl)
11	16.227	3.45	1,2-benzenedicarboxylic acid and diethyl ester
12	17.505	24.29	9-octadecenoic acid (Z) and oleic acid
13	17.911	2.38	Cis-9,cis-12-octadecadienoic acid, and cis-linoleic acid
15	19.574	1.28	Cyclopropanebutanoic acid,
16	20.129	6.88	Tetradecanoic acid and myristic acid
17	20.804	1.21	1H-purine-6-amine
18	23.507	4.74	Hexadecanoic acid and palmitic acid
19	28.284	30.62	1H-purine-2,6-dione and 3,7-dihydro-1,3,7-trimethyl
(b) Sanding, 40 min
1	9.76	0.19	Dodecanoic acid, methyl ester, and lauric acid
3	12.025	4.83	Tetradecanoic acid and methyl ester
4	13.323	0.26	Phenol and 2-methoxy-4-(2-propenyl)
6	14.613	16.61	Hexadecanoic acid and methyl ester
7	15.887	1.04	Phenol, 2-methoxy-4-(1-propenyl), phenol, and 2-methoxy-4-propenyl
8	16.026	0.36	1H-purine-6-amine
9	16.226	3.14	1,2-benzenedicarboxylic acid and diethyl ester
10	17.364	0.27	Octadecanoic acid and methyl ester
11	17.512	26.58	9-octadecenoic acid, methyl ester, and oleic acid
12	17.797	0.29	9-octadecenoic acid and octadec-9-enoic acid
13	17.927	2.82	Methyl octadeca-9,12-dienoate, and octadeca-9,12-dienoic acid
15	19.58	0.83	Cyclopropanebutanoic acid
16	20.066	6.93	Tetradecanoic acid and myristic acid
17	20.804	0.95	1H-purine-6-amine
18	23.403	8.19	n-Hexadecanoic acid, hexadecanoic acid, n-Hexadecoic acid, and palmitic acid
19	23.8	0.94	1H-purine-6-amine
20	28.253	18.69	1H-purine-2,6-dione and 3,7-dihydro-1,3,7-trimethyl
21	29	0.43	1H-purine-6-amine
21	29.534	5.07	Oleic Acid, 9-octadecenoic acid, and delta.(Sup9)-cis-oleic acid
(c) Sanding, 120 min
3	9.759	0.24	Tridecanoic acid and methyl ester
5	12.02	7.71	Methyl ester, myristic acid, methyl ester, and methyl myristate
6	13.325	0.19	Phenol, 2-methoxy-4-(2-propenyl), 4-allyl-2-methoxy-phenol, and 1,3,4-eugenol
8	14.602	22.22	Hexadecanoic acid, methyl ester, palmitic acid, and methyl ester
9	16.005	0.67	1H-purine-6-amine, [(2-fluorophenyl)methyl]
11	17.347	2.12	Octadecanoic acid, methyl ester, stearic acid, and methyl ester
12	17.506	38.18	9-octadecenoic acid, methyl ester, and oleic acid
13	17.92	1.81	10,13-octadecadienoic acid and methyl ester
14	18.926	2.01	1H-purine-6-amine and [(2-fluorophenyl)methyl]
15	19.579	1.72	Cyclopropanebutanoic acid and methyl ester
16	20.493	2.37	9-octadecenoic acids
18	23.787	1.79	1H-purine-6-amine and [(2-fluorophenyl)methyl]
19	28.278	5.16	Cafeina, caffein, caffine, cafipel, coffeine, guaranine, and koffein

Table 3. Relative percentages of some major compounds present in both types of samples

Tea type	Compounds present	20-min	40-min	120-min
Sanding	Myristic acid	6.88^a ± 0.07	6.93^a ± .09	7.71^b ± 0.08
	palmitic acid	13.98^a ± 0.13	16.61^b ± 0.17	22.22^c ± 0.12
	Stearic acid	^*	0.27^a ± 0.12	2.12^a ± 0.06
	oleic acid	24.29^a ± 0.22	26.58^b ± 0.06	38.18^c ± 0.12
	Linoleic acid	2.38^b ± 0.10	2.82^c ± 0.12	1.72^a ± 0.04
	Caffein	30.62^c ± 0.22	18.69^b ± 0.12	5.16^a ± 0.22
	1H-purin-6-amine	1.21^a ± 0.02	1.21^a ± 0.03	1.91^b ± 0.07
Leaf grade	Myristic acid	1.64^b ± 0.07	3.23^c ± .22	1.42^a ± 0.06
	palmitic acid	24.43^c ± 0.23	18.31^a ± 0.06	19.22^b ± 0.08
	Stearic acid	2.24^c ± 0.07	0.57^a ± 0.03	1.61^b ± 0.07
	oleic acid	36.94^c ± 0.12	31.24^a ± 0.08	33.09^b ± 0.32
	Linoleic acid	1.95^b ± 0.08	5.57^c ± 0.22	1.43^a ± 0.31
	Caffein	7.42^a ± 0.09	18.08^b ± 0.05	17.8^b ± 0.19
	1H-purin-6-amine	2.1^a ± 0.09	1.95^a ± 0.05	1.76^a ± 0.13

Notes: The values are mean ± SD of three independent determinations.

Means followed by different superscripts within a row are significantly (p ≤ 0.05) different.

^* Not detected.

Table 4. Biological activity of some phytocomponents identified in the green extracts

Compound name	Bioactivity	References
Methyltetradecanoate/Myristic acid	Antioxidant, Cancer-preventive, Hypercholesterolemic, Lubricant, Nematicide	Gopalakrishnan and Vadivel (2011)
Hexadecanoic acid/Palmitic acid	Antioxidant, Hypocholesterolemic Nematicide, Pesticide, Anti androgenic, Flavor, Hemolytic,5-Alpha reductase inhibitor	Gopalakrishnan and Vadivel (2011)
Octadecanoic acid/Stearic acid	5-Alphareductase inhibitor, Cosmetic, Flavor, Hypocholesterolemic	Gopalakrishnan and Vadivel (2011)
9-octadecenoic acid/Oleic acid	Antiinflammatory, Antiandrogenic Cancer preventive, Dermatitigenic Hypocholesterolemic, 5-Alpha reductase inhibitor, Anemiagenic Insectifuge, Flavor	Jananie, Priya, and Vijayalakshm (2001)
9,12-octadecadienoic Acid/Linoleic acid	Hypocholesterolemic, Nematicide Antiarthritic, Hepatoprotective Anti androgenic, Hypocholesterolemic Nematicide, 5-Alpha reductase inhibitor, Antihistaminic, Anticoronary Insectifuge, Antieczemic, Antiacne Anticancerous and diuretic	Mathew (2011), Nehlig, Daval, and Debry (1992)
1H-purine-2,6-dione/Caffein	Prevents: liver cirhossis, Type 2 diabetes, apnea of prematurity, bronchopulmonary dysplasia in premature infants.Secondary metabolic products of caffeine: paraxanthin increase lipolysis, theobromine is a vasodilator, theophylline acts as a muscle relaxant, anti asthmatic.	Kugelman and Durand (2011), Van (2008), Muriel and Arauz (2010)

Table 5. Effect of extraction time on total phenolic content (TPC) and antioxidant activity of two green tea types

Time	Leaf-grade type		Sanding type
	TPC (GAE/g)	DPPH (% inhibition)	TPC (GAE/g)	DPPH (% inhibition)
20	120.79^b ± 0.69	73.69^b ± 0.31	116.59^c ± 1.3	72.55^b ± 1.2
40	137.59^a ± 1.56	78.34^a ± 0.94	131.37^a ± 1.2	76.4^a ± 0.62
60	118.71^b ± 0.48	74.13^b ± 1.0	127.45^b ± 0.96	74.45^ab ± 1.3

Notes: The values are mean ± SD of three independent determinations.

Means followed by different superscripts within a column are significantly (p ≤ 0.05) different.

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