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GC-FID guided Identification and Quantification of detectable Phytochemicals in selected Commercial Chamomile Herbal Tea

Article Information

Kenneth C. Ugoeze1, Nora Amadi1, Ngozi A. Okoronkwo2, Sunday O. Abali1, Kennedy E. Oluigbo3, Bruno C. Chinko*, 4

1Department of Pharmaceutics & Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, University of Port Harcourt, Port Harcourt, Nigeria.

2Department of Zoology & Environmental Biology, Michael Okpara University of Agriculture, Umudike, Abia State, Nigeria.

3Department of Clinical Pharmacy and Biopharmaceutics, Faculty of Pharmaceutical Sciences, Enugu State University of Science and Technology, Agbani City, Enugu, Nigeria.  

4Department of Human Physiology, Faculty of Basic Medical Sciences, University of Port Harcourt, Port Harcourt, Nigeria.

*Corresponding author: Bruno C. Chinko, Department of Human Physiology, Faculty of Basic Medical Sciences, University of Port Harcourt, Port Harcourt, Nigeria

Received: 23 December 2022; Accepted: 05 January 2023; Published: 14 January 2023

Citation: Kenneth C. Ugoeze, Nora Amadi, Ngozi A. Okoronkwo, Sunday O. Abali, Kennedy E. Oluigbo, Bruno C. Chinko. GC-FID guided Identification and Quantification of detectable Phytochemicals in selected Commercial Chamomile Herbal Tea. International Journal of Applied Biology and Pharmaceutical Technology 14 (2023): 01-11.

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Abstract

Background and objective: Phytochemicals are plant-based bioactive ingredients found in tea, fruits, and vegetables with multiple health benefits. This study aimed to identify and quantify the detectable phytochemicals in selected products of chamomile herbal tea (CHT) using the Gas Chromatography - Flame Ionization Detector (GC-FID) approach.

Methods: Selected CHTs were coded as CHT-A to CHT-G. CHT-F was crude and unbranded however, CHT-G was an oil extract of CHT. Ethanolic extracts of CHT were analysed and quantified for their phytoconstituents using the GC-FID method.

Results: Phytochemicals detected in their order of abundance in most CHTs were flavonoids > glycosides > alkaloids > steroids > anti-nutrients > saponins > tannins > resveratrol. The flavonoids comprised rutin > flavanone > flavone > anthocyanin > epicatechin > kaempferol > naringenin > proanthocyanin, etc. where the glycosides found in each CHT involved cardiac and cyanogenic glycosides. Alkaloids were highest in CHT-A as ribalinidine > spartein > lunamarin with ephedrine detected just in CHT-B, CHT-C, CHT-D, CHT-E and CHT-F. Saponins occurred in four samples mainly as sapogenin. Anti-nutrients were detected in each CHT and included the phytates and oxalates. Resveratrol was detected in three samples only.

Conclusion: Most of the CHTs evaluated showed the presence of flavonoids, glycosides, alkaloids, steroids, anti-nutrients, saponins, tannins and resveratrol.

Keywords

GC-FID identification and quantification, Phytochemical, Chamomile, Herbal tea

GC-FID identification and quantification articles, Phytochemical articles, Chamomile articles, Herbal tea articles

Article Details

1. Introduction

Phytochemicals are considered plant-based bioactive chemicals with various therapeutic and nutritious benefits [1-3]. They are classified as primary (PM) or secondary metabolites (SM). The PM comprises the common sugars, amino acids, proteins, purines and pyrimidines of nucleic acids, chlorophylls, etc. whereas the SM are the plant chemicals such as alkaloids, terpenes, flavonoids, lignans, plant steroids, saponins, phenolics, glycosides, etc. [1].   Due to their antimicrobial, anti-inflammatory, anthelmintic, anti-carcinogenic, anti-genotoxic, anti-proliferative, anti-mutagenic, anti-allergic, immune-modulatory and anti-oxidative properties, the SM could play protective roles against pathogens or damaging disorders [4-9]. There are over 8000 natural flavonoids [5] with their health-based biotic actions ascribed to their antioxidant influence [6, 7]. The antioxidant and anti-inflammatory actions of the flavonoids aid in toxin-mediated stress and protracted infection inhibition [10]. Flavonoid-rich foods include all nourishments of plant basis, predominantly tea, fruit, vegetables, grains, legumes, nuts, and wine [11, 12].

Tea is processed from the vegetative parts of Camellia sinensis [13]. They are used as a medicinal drink by two-thirds of the globe [14] and have been designated as “safe” by the US Food and Drug Administration's (FDA) list of compounds generally recognized as safe (GRAS) [15]. Normally, Tea refers to dried leaves processed from leaves of Camellia sinensis (true tea).  Other infusions got from parts of other suitable plants are also noted as tea, but, are called tisanes or herbal tea, hence, there are true or traditional tea and herbal tea [12, 13]. True tea is grouped as green, black/dark, white, yellow and oolong which is established on the extent of oxidation of the leaves [16]. Green tea is a non-fermented tea [17], they are processed from freshly harvested vegetation of the tea plant without fermentation after withering, steaming or pan firing, drying and grading to retain its content of polyphenols [15, 16, 18]. On the other hand, black tea is wholly fermented tea, processed with polyphenol oxidase converting its polyphenols into a series of new products like theaflavins, theaflagallins and thearubigens obtainable in black tea  [16, 18]. Oolong tea is a semi-fermented tea gotten when leaves are wilted in the sun and moderately battered to attain brief partial oxidation resulting in an intermediate product, the oolong tea [18-20].

Herbal tea or tisanes implies any infusions taken as a drink derived from other parts of vegetation other than Camellia sinensis [6, 15]. They are often caffeine free compared to traditional teas. It could be gotten from plants like chamomile, rose hip, ginger, turmeric, valerian, hibiscus, peppermint, etc. [21]. Chamomile is one of the early therapeutic herbs with standardized forms prepared from dried flowers of Matricaria species and of the daisy family, Asteraceae/ Compositae [22]. Two common varieties are the German Chamomile (Matricaria chamomilla) and Roman Chamomile (Chamaemelum nobile) [23]. Their preparations are used to manage several ailments like hay fever, inflammation, muscle spasms, menstrual disorders, insomnia, ulcers, wounds, gastrointestinal disorders, rheumatic pain and haemorrhoids while their essential oils are applied widely in cosmetics and aromatherapy [24, 25]. Their therapeutic influence is largely based on their dried flowers' terpenoid and flavonoid content [24]. Chamomile herbal tea is essentially caffeine-free and is believed to be a calming and soothing effect which promotes sleep and reduces anxiety [26, 27]. They have also been found to be effective in treating injuries, ulcers, eczema, gout, skin irritations, bruises, burns, canker sores, neuralgia, sciatica, rheumatic pain, haemorrhoids and mastitis [25, 28]. Aside from their use as tea, and tincture, they are used as poultices and tincture [29]. Some of the flavonoids found in chamomile are apigenin, quercetin, and patuletin [30, 31]. Apigenin is known to bind to benzodiazepine receptors in the brain to enhance sleep and diminish insomnia [32, 33]. Similarly, their essential oils are used in aromatherapy to enhance sleep and relieve anxiety [34].

Despite the aforementioned benefits and safety margin of chamomile, there has been no confirmation of their reaction to other medications or their safety among children and pregnant women. Therefore, the present study aims to identify and quantify the phytochemicals in selected commercial chamomile herbal tea using Gas Chromatography with flame ionization detection (GC-FID) methods.

2. Materials and Methods

2.1 Procurement of Samples

Five brands of chamomile herbal tea (CHT) were randomly sourced from retail outlets in Port Harcourt, Nigeria and coded CHT-A, CHT-B, CHT-C, CHT-D and CHT-E. Also, a crude unpackaged dry chamomile flower was sourced from a retail outlet in Abuja and coded as CHT-F while its oil extract was coded as CHT-G.

2.1 Preparation of samples and extraction of phytoconstituents

A teabag of each CHT was macerated in 200 ml of ethanol (Honeywell, Germany) for 48 hours. It was filtered and evaporated to obtain the extract. A 0.1 g of the extract was re-extracted in 25 ml of ethanol in a test tube immersed in a water bath (60 o C) for 90 mins. It was transferred to a separatory funnel. The test tube was washed in steps with 20 ml of ethanol, 10 ml of cold water, 10 ml of hot water and 3 ml of n-hexane (BDH, England) respectively and transferred into the separatory funnel. The extracts were pooled and washed thrice with 10 ml of 10 % v/v ethanol-aqueous solution. The solution was dried with anhydrous sodium sulphate (Sigma-Aldrich, USA) and the solvent was evaporated. The sample was solubilized in 1000 ul of pyridine (Sigma-Aldrich, USA), of which 200 ul was transferred to a vial for analysis [35, 36].

2.2 Quantification of the phytoconstituents by GC-FID

The analysis of phytoconstituents was carried out on a BUCK M910 Gas Chromatography fitted with an HP-5MS column (30 m in length × 250 μm in diameter × 0.25 μm in thickness of film). Spectroscopic detection by GC–FID involved an electron ionization system which used high-energy electrons (70 eV). Pure helium gas (99.995 %) was employed as the carrier gas with a flow rate of 1 mL/min. The early temperature was set at 50 –150 °C with an increasing rate of 3 °C/min and a holding time of about 10 min. Later the temperature was amplified to 300 °C at 10 °C/min. A 1µl of the prepared 1% of the extracts diluted with respective solvents was injected in a splitless mode. The relative quantity of the chemical compounds present in each of the extracts was expressed as a percentage based on the peak area produced in the chromatogram [36, 37]. Bioactive compounds extracted from the respective batches of extracts were identified based on the GC retention time on the HP-5MS column and matching the spectra with the computer software standards data (Replib and Mainlab data of GC–FID systems).

3. Results

3.1 GC-FID Quantified Phytoconstituents of selected Chamomile Herbal Tea

Table 1: Composition of the phytochemical constituents detected in CHT-A

Phytochemical

Component

Retention

Area

Height

Conc. (ug/mol)

% Composition

Flavonoids (48.76%)

Naringin

0.873

4824.9767

377.473

3.7794

5.42

Anthocyanin

5.483

2954.8422

231.937

2.6609

3.81

Flavan-3-ol

7.873

4968.2281

389.42

2.886

4.14

Naringenin

14.036

3250.6604

255.555

2.7644

3.96

Rutin

19.973

4928.684

387.165

3.976

5.7

Flavonones

20.616

3419.6849

268.983

2.6396

3.78

Kaempferol

25.96

7289.2832

570.18

5.721

8.2

Flavone

34.15

3084.5871

241.227

2.6798

3.84

Epicatechin

36.773

4449.9298

348.939

5.1499

7.38

Catechin

38.64

8024.0488

624.601

1.7616

2.53

Alkaloids (13.25%)

Ribalinidine

10.83

6003.0656

470.455

4.9682

7.12

Lunamarin

0.386

2631.0434

206.27

1.1557

1.66

Spartein

43.536

5031.2166

394.788

3.123

4.48

Glycoside (2.11%)

Cardiac glycoside

3.093

3389.3006

265.8

1.4722

2.11

Tannin (2.54%)

Tannin

9.35

3057.1151

240.132

1.7753

2.54

Steroids (12.95%)

Steroids

23.363

6954.4788

536.637

9.0361

12.95

Anti-nutrients (20.38)

Phytate

29.326

5273.2844

410.247

5.3158

7.62

Oxalate

31.653

6802.6676

532.675

8.9007

12.76

Table 2: Composition of the phytochemical constituents detected in CHT-B

Phytochemical

Component

Retention

Area

Height

Conc. (ug/ml)

% Composition

Flavonoids (61.18%)

Proanthocyanin

0.113

1472.1354

410.405

0.6902

0.71

Proanthocyanin

0.17

5680.6418

256.735

2.6632

2.75

Anthocyanin

3.946

8222.3654

131.434

7.4044

7.66

Flavan-3-ol

6.893

4472.4692

71.805

2.598

2.69

Naringenin

13.3

4660.7432

77.1

3.9636

4.1

Rutin

15.783

12553.4794

203.066

10.1269

10.47

Flavonones

19.573

12438.6034

195.876

9.601

9.93

Kaempferol

26.003

6617.3268

107.594

5.1937

5.37

Flavone

34.073

9054.95

127.289

7.8665

8.13

Epicatechin

37.363

5799.3524

113.423

6.7116

6.94

Catechin

38.293

10686.5711

156.747

2.3461

2.43

Alkaloids (10.38)

Riblinidine

10.59

4269.5216

68.918

3.5335

3.65

Ephedrine

42.72

11673.1373

120.423

6.5073

6.73

Saponins (2.16%)

Sapogenin

40.976

2989.3324

57.369

2.0892

2.16

Steroids (6.23%)

Steroids

22.29

4635.9377

75.497

6.0235

6.23

Glycosides

Cyanogenic glycoside

28.653

10030.8184

98.828

14.8305

15.34

18.48%

Cardiac glycoside

2.22

7007.3973

110.755

3.0439

3.15

Other Phenols

Resveratrol

39.563

3987.0637

71.359

1.5148

1.57

1.57%

Table 3: Composition of the phytochemical constituents detected in CHT – C

Phytochemical

Component

Retention

Area

Height

Conc. (ug/ml)

% Composition

Flavonoids (46.89%)

Proanthocyanin

0.086

469.529

179.446

0.2553

0.18

Anthocyanin

4.063

4293.166

73.064

3.8661

2.65

Flavan-3-ol

6.07

27561.7994

195.841

16.0103

10.99

Flavonones

12.966

5856.362

62.803

4.5203

3.1

Naringenin

15.46

4561.1764

49.349

3.9752

2.73

Rutin

17.966

11050.1446

115.177

7.3233

5.03

Catechin

20.313

12511.0674

129.573

3.0213

2.07

Kaempferol

25.683

9599.0294

103.696

7.5339

5.17

Flavone

33.03

20029.9992

149.673

12.4294

8.53

Epicatechin

36.88

6804.3615

71.638

9.3826

6.44

Alkaloids (5.68%)

Lunamarin

0.173

3559.1162

92.448

1.5633

1.07

Epihedrine

42.4

2720.3349

37.666

1.2832

0.88

Spartein

44.16

10938.5209

109.349

5.4319

3.73

Saponins (4.90%)

Sapogenin

39.196

10225.2962

105.22

7.1464

4.9

Steroids (10.05%)

Steroids

22.726

9150.8282

96.755

14.6366

10.05

Glycosides (15.41%)

Cardiac glycoside

2.413

13711.4174

130.202

5.3774

3.69

Cyanogenic glycoside

27.513

11552.32

118.637

17.08

11.72

Anti-nutrients

Oxalate

29.853

5038.285

54.96

5.1795

3.55

17.06%

Phytate

10.366

19525.9631

201.089

19.6834

13.51

Table 4: Composition of the phytochemical constituents detected in CHT-D

Phytochemical

Component

Retention

Area

Height

Conc. (ug/ml)

% Composition

Flavonoids (43.17%)

Proanthocyanin

0.086

357.1588

179.448

0.2679

0.16

Naringin

0.233

2989.6825

117.856

2.7796

1.62

Anthocyanin

4.12

6154.4164

144.253

5.5421

3.23

Flavan-3-ol

6.016

18053.9923

409.964

13.1092

7.65

Flavonones

12.97

6235.5473

141.356

4.813

2.81

Naringenin

15.46

4965.0218

112.527

4.2223

2.46

Rutin

17.966

11330.0842

256.503

8.2963

4.84

Catechin

20.313

12744.4217

285.881

3.6373

2.12

Flavone

32.996

14335.5145

323.847

12.4541

7.27

Kaempferol

25.65

991 7.8566

226.671

7.7841

4.54

Epicatechin

36.876

6977.1841

158.007

11.0654

6.46

Lunamarin

0.16

539.1959

116.491

0.2368

0.14

Alkaloids (12.33%)

Ephedrine

42.276

3426.0155

78.72

1.9099

1.11

Spartein

44.17

10459.4821

237.898

8.4403

4.93

Ribalinidine

7.47

8392.9472

191.094

10.5452

6.15

Saponins (4.17%)

Sapogenin

39.2

10228.6703

231.342

7.1488

4.17

Glycosides (13.40%)

Cyanogenic glycoside

27.536

11365.4653

259.34

16.8038

9.81

Cardiac glycoside

2.39

12099.0446

277.929

6.1565

3.59

Steroids (8.93%)

Steroids

22.73

9568.19

216.422

15.3042

8.93

Other Phenolics (2.28%)

Resveratol

34.6

6049.213

137.14

3.9072

2.28

Anti-nutrients (15.71%)

Oxalate

29.86

5472.7816

124.058

7.1606

4.18

Phytate

10.366

19594.4088

442.508

19.7524

11.53

Table 5: Composition of phytochemical constituents detected in CHT-E

Phytochemical

Component

Retention

Area

Height

Conc. (ug/ml)

% Composition

Flavonoids

Proanthocyanin

0.116

3681.8254

411.025

3.4523

3.51

57%

Anthocyanin

3.95

8180.0436

637.037

8.4186

8.56

Flavan-3-ol

6.893

4491.1913

350.845

2.6089

2.65

Naringenin

13.3

4918.608

385.135

4.4167

4.49

Rutin

15.783

12794.3857

919.8

7.9394

8.07

Flavonones

19.516

12631.1433

566.996

9.7496

9.91

Kaempferol

26

6833.3794

529.584

4.7323

4.81

Flavone

34.206

5932.5289

458.609

5.1539

5.24

Epicatechin

37.26

6525.2532

508.857

7.5517

7.68

Catechin

38.326

9393.7324

727.257

2.4747

2.52

Alkaloids (10.60)

Ephedrine

42.086

6000.1311

470.143

3.3448

3.4

Spartein

42.943

6524.2634

510.968

4.0498

4.12

Ribalinidine

10.593

4339.0384

337.681

3.0329

3.08

Steroids (6.27%)

Steroids

22.293

4749.7578

372.508

6.1714

6.27

Glycosides (12.71%)

Cyanogenic glycoside

28.566

5744.9478

450.313

8.4939

8.63

Cardiac glycoside

2.223

6793.2211

528.999

4.0047

4.07

Other Phenolics (1.70%)

Resveratol

39.586

4412.4024

345.673

1.6764

1.7

Saponins (2.45%)

Sapogenin

40.93

3451.568

270.978

2.4123

2.45

Anti-nutrients  (8.83%)

Phytate

29.493

4459.3978

349.793

4.4954

4.57

Oxalate

33.753

3207.2732

252.912

4.1939

4.26

Table 6: Composition of the phytochemical constituents detected in CHT-F

Phytochemical

Component

Retention

Area

Height

Conc. (ug/ml)

% Composition

Flavonoids (42.46)

Proanthocyanin

0.19

5184.3944

427.802

2.4306

1.72

Naringin

1.583

4709.7496

369.572

4.3788

3.09

Anthocyanin

3.55

3903.4112

306.58

3.5151

2.48

Flavan-3-ol

4.4

10229.5051

797.09

7.4278

5.24

Flavonones

12.99

7261.1404

564.292

5.6047

3.96

Naringenin

15.62

5351.2845

419.38

4.5508

3.21

Rutin

18.95

6368.0202

498.244

4.6629

3.29

Flavone

35.65

17427.5578

1329.989

15.1403

10.69

Epicatechin

36.526

5159.9954

404.908

5.9717

4.22

Kaempferol

25.563

4875.0349

382.357

3.8262

2.7

Catechin

28.276

9186.5206

716.488

2.6218

1.85

Alkaloids (7.39%)

Ephedrine

42.706

13247.6644

1026.936

7.3851

5.21

Ribalinidine

13.973

3725.9862

292.76

3.0837

2.18

Tannins (2.22%)

Tannin

13.273

5414..6802

430.677

3.1444

2.22

Steroids (7.83%)

Steroids

22.456

8539.7226

666.846

11.0958

7.83

Glycoside

Cardiac glycoside

2.633

12170.5138

945.575

6.1929

4.37

18.70%

Cyanogenic glycoside

27.91

13725.1531

1063.342

20.2926

14.33

Anti-nutrients

Oxalate

33.81

18147.5364

1384.596

23.7444

16.77

21.40%

Phytate

12.62

6505.2012

510.587

6.5577

4.63

Table 7: Composition of the phytochemical constituents detected in CHT-G

Phytochemical

Component

Retention

Area

Height

Conc. (ug/ml)

% Composition

Flavonoids (49.61%)

Naringin

0.763

5779.5425

98.904

5.3734

8.11

Naringenin

14.036

3240.2268

52.096

2.7555

4.16

Rutin

20.17

8342.6846

101.927

6.1088

9.22

Kaempferol

25.956

7126.6852

115.871

5.5934

8.44

Anthocyanin

5.48

2719.094

45.696

2.4486

3.7

Flavone

34.15

2915.6571

48.251

2.533

3.82

Epicatechin

36.643

7925.2515

127.307

2.2619

3.41

Catechin

38.643

7925.2515

127.307

2.2619

3.41

Flavan-3-ol

7.876

4852.1955

78.442

3.5232

5.32

Alkaloids (12.23%)

Ribalinidine

10.83

6000.1402

95.538

4.9658

7.5

Spartein

43.533

5050.4626

80.811

3.135

4.73

Tannins (2.48%)

Tannin

9.346

2829.2458

48.179

1.643

2.48

Glycosides

Cardiac glycoside

3.093

3039.8329

51.73

1.5468

2.34

2.34%

Steroids  (13.12%)

Steroids

23.366

6688.5282

108.88

8.6905

13.12

Anti-nutrient (20.22%)

Phytate

29.33

4936.9085

81.904

4.9767

7.51

Oxalate

31.65

6434.1545

106.616

8.4185

12.71

4. Discussions

The results obtained from the GC-FID of selected chamomile herbal tea and dry chamomile flower and essential oils are summarised in Tables 1 – 7. These results are discussed in this section under flavonoids, alkaloids, glycosides, saponins, tannins, steroids, anti-nutrients and other phenols such as resveratrol.

4.1 Flavonoids

The level of flavonoids detected in the various CHT were as CHT-B > CHT-E > CHT-G > CHT-A > CHT-C > CHT-C > CHT-D > CHT-F. The types of flavonoids identified include proanthocyanin, anthocyanin, flavan-3-ol, naringenin, rutin, flavanones, kaempferol, flavone, catechin, epicatechin, etc. (Tables 1-7). The variation in the composition of these flavonoids from one CHT to the other may be attributed to differences in environment or influences like latitude, longitude, rainfall, temperature and soil quality [38]. Given these differences, and with CHT-B presenting the highest concentration of flavonoids displayed its subgroup of flavonoids as Rutin   > flavonone > flavone > anthocyanin > epicatechin > kaempferol > naringenin > proanthocyanin, etc. (Table 2). Flavonoids have been known to possess immense pharmacological benefits such as anti-oxidative, anti-mutagenic, anti-inflammatory, anti-carcinogenic, antitumour, anti-HIV, antidiarrhoeal, antihepatotoxic, antifungal, antilipolytic, vasodilator, immunostimulant and anti-ulcerogenic properties and enzyme modulatory functions [39, 40]. Rutins have been found in fresh leaves, red wine and tea [12, 41, 42]

4.2 Alkaloids

The composition of alkaloids detected was CHT-A > CHT-D > CHT-G > CHT-E > CHT-B > CHT-F > CHT- C. The various forms of alkaloids detected were ribalinidine, lunamarin, spartein, ephedrine, etc., though, ephedrine was not detected in CHT-A and CHT-G.  Alkaloids are natural products that have heterocyclic nitrogen atoms [43-45]. They found use in the ancient preparation of spices, drugs and poisons. Lunamarin retains anticancer, immunomodulatory, anti-estrogenic and anti-amoebic activities [46] while ribalinidine possesses a radical scavenging influence.

4.3 Glycosides

The composition of glycosides identified in the various CHT were CHT-F > CHT-B > CHT-C > CHT-D > CHT-E > CHT-G > CHT-A with cardiac and cyanogenic glycosides prevailing (Tables 1-7). With the highest level of glycosides detected in CHT-F, the extent of cardiac and cyanogenic glycosides was 4.37 and 14.33 % respectively.  In CHT-B, they were 3.15 and 15.34 % correspondingly whereas it was 3.69 and 11.72 % in CHT-C. Further, it was 3.59 and 9.81% in CHT-D while in CHT-E it was 4.07 and 8.63 %. Only cardiac glycoside was detected in CHT-A and CHT-G at 2.11 and 2.34 % respectively. The cyanogenic glycosides retained higher concentrations where it was detected. Glycosides are plant-based substances comprising of a glucose unit confined to an aglycone like alcohol, phenol or steroid nucleus through a glycosidic bond [47] with potent antibacterial, antifungal, anti-inflammatory, antioxidant, antiviral and anticancer activities [48, 49]. Cardiac glycosides are used in the treatment of cardiac insufficiency [48, 50] by increasing the output force of the heart and decreasing its rate of contractions by inhibiting the cellular Sodium-Potassium-ATPase pump [50]. However, their relative toxicity prevents their extensive application [51]. On the other hand, cyanogenic glycosides which are mostly found in foods including linamarin, amygdalin and prunasin [52] are known to release hydrogen cyanide when chewed or digested [53] resulting in significant cyanide poisoning. However, processing methods, such as peeling, drying, grinding, soaking and fermentation, boiling or cooking have been reported to cause a significant reduction in the cyanogenic glycosides of processed foods [54].

4.4 Saponins

The saponin detected was sapogenin which occurred in these CHTs as CHT-C (4.90 %) > CHT-D (4.17 %) > CHT-E (2.45 %) > CHT-B (2.16 %) (Tables 1-7). Saponins constitute a vast group of glycosides occurring in many plants and are characterized by their surfactant properties. They are grouped as triterpenoid and steroid saponins [55].  The steroidal saponins are essential precursors for steroid drugs, comprising anti-inflammatory agents, androgens, oestrogens and progestins [56]  while triterpene saponins exhibit various pharmacological activities, including anti-inflammatory, molluscicidal, antitussive, expectorant, analgesic and cytotoxic influences and include the ginsenosides, which are responsible for some of the pharmacological activity of ginseng and the active triterpenoid saponins from liquorice [57, 58].

4.5 Tannins

Tannins were detected only in these CHTs as CHT-A (2.54 %) > CHT-G (2.48 %) > CHT-F (2.22 %). The existence of tannins in tea leaves accounts for the bitter and dry sensation felt when tea is tasted. Tannins are higher in black tea than in oolong, green and white teas [59]. Tannins also occur in red wine, coffee, grapes, apple juice, strawberry, raspberry, blackberry, pomegranate, plums, walnuts, olives, chickpeas, lentils, chocolate and cocoa [60].  Foods rich in tannins have been considered to be of low nutritional value since tannins have been reflected as an anti-nutrient, due to their ability to decrease the efficiency in converting the absorbed nutrients to relevant substances [61, 62]. Tea polyphenols and several components of tannin have been suggested as anti-carcinogenic and many tannin molecules have also been shown to reduce the mutagenic activity of several mutagens. These properties have been attributed to their anti-oxidative properties which enable them to defend against oxidative impairment [61].

4.6 Anti-nutrients

The ‘anti-nutrients’, comprise lectins, oxalates, phytates, phytoestrogens and tannins [63]. Phytates and oxalates were detected in the CHTs as CHT-F > CHT-A > CHT-G > CHT-C > CHT-D > CHT-E (Tables 1, 3-7). Oxalate was higher than phytate in CHT-F (16.77 %), CHT-A (12.76 %) and CHT-G (12.71 %) whereas phytate was higher than oxalate in CHT-C (13.51 %), CHT-D (11.53 %) and CHT-E (4.57 %). Anti-nutrients limit the bioavailability of vital nutrients by binding to vital micronutrients which prevents the body from absorbing them or hindering the peak effects of some digestive enzymes, thereby, inhibiting the appropriate breakdown of food [63]. For example, oxalates are known to affect calcium absorption and use by forming calcium oxalate crystals which could lead to kidney stones. They also irritate and cause swelling in the mouth and throat, and are capable of forming tissue crystals leading to indications of arthritis [64]. Some of the health benefits derivable from dietary phytate include anti-cancer, anti-calcification, antioxidant, antihyperglycaemic and hypolipidaemic activities [63, 65]. They can bind to harmful trace elements like lead and cadmium thereby reducing their bioavailability. It has been associated with certain health benefits, including blood glucose – and lipid-lowering effects, anticancer activity, antioxidant properties, and anti-calcification. The ability of phytate to bind toxic trace elements such as cadmium and lead and reduce their bioavailability has been documented [66].

4.7 Steroids

Steroids were detected in the CHTs, but, due to the limitations of the study, the type of steroid identified was not identified. However, steroids were detected in the order CHT-G (13.12 %) > CHT-A (12.95 %) > CHT-C (10.05 %) > CHT-D (8.93 %) > CHT-F (7.83 %) > CHT-E (6.27 %) > CHT-B (6.23 %) (Tables 1-7). Amongst the plant-based steroids, phytosterols are the most abundant [67]. They are known to reduce blood cholesterol by inhibiting intestinal absorption of cholesterol thereby reducing the risk of heart attack and stroke [67]. They have also been demonstrated to slow the in vitro development and progression of various cancers [68].

4.8 Resveratrol

Resveratrol, a non-flavonoid polyphenol was detected in low levels in CHT-D (2.28 %) > CHT-E (1.70 %) > CHT-B (1.57 %). It is a polyphenolic phytoalexin formed by plants like grapes, peanuts and berries and retains anti-inflammatory, antioxidant, antiplatelet, anticancer and anti-diabetic activities [69, 70]. In vitro investigations have also revealed its ability to prevent all phases of carcinogenesis comprising initiation, promotion and progression at lower doses [71]. However, at higher doses, resveratrol acts as a pro-apoptotic compound which signals the death of cancer cells. They are also able to depress cardiac function [71].

5. Conclusion

The GC-FID-guided phytochemical identification and quantification of selected CHTs showed that they contain mostly detectable flavonoids, alkaloids, glycosides, saponins, tannins, steroids, anti-nutrients and other phenols such as resveratrol. This present study corroborates the literature on the abundant phytochemical constituents of chamomile which serves as the basis for the numerous health benefits ascribed to chamomile herbal tea.

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