Anise (Pimpinella anisum L.), a dominant spice and traditional medicinal herb for both food and medicinal purposes

Abstract

Aromatic plants such as anise seed have a long traditional use in both folk and conventional medicine and of course in the pharmaceutical industry. Important compounds found in anise seed include estragol, p-anisaldehyde, anise alcohol, acetophenone, pinene, and limonene, but the most important volatile oil that gives the characteristic sweet, aromatic flavor to seeds is anethole. The recent studies have shown that anise seeds and essential oil have antioxidant, antibacterial, antifungal, anticonvulsant, anti-inflammatory, analgesic, gastro-protective, antidiabetic, and antiviral activities. Other important benefits of anise seeds are stimulant, carminative, expectorant, insecticide, vermifuge, digestive, antispasmodic, antirheumatic, antiseptic, antiepileptic, antihysteric, culinary significance, keeps the heart strong by its importance role to control the blood pressure, one of the best gas-releasing agent, eases many hormonal problems in females, hair benefits, skin benefits, and it may reduce symptoms of depression. Anise seed and its extract also use in savory dishes, baked goods, and different drinks in both ancient and modern time. Anise seeds are good source of many essential B-complex vitamins such as pyridoxine, niacin, riboflavin, and thiamin. The seeds are also important source of minerals like calcium, copper, potassium, iron, manganese, magnesium, and zinc. Antioxidant vitamins such as vitamin C and A can also be found in the spice. More clinical studies are necessary to uncover the numerous substances and their effects in ginseng that contribute to public health.

Keywords:

• Health benefits
• traditional Chinese medicine
• traditional Asian medicine
• clinical aspect
• anethole

PUBLIC INTEREST STATEMENT

Pimpinella asnisum L. is an aromatic, annual grassy herb with 30–50 cm height, white flowers and small green to yellow seeds with secondary feather like leaflets of bright green, twice pinnate. Keeping the traditional knowledge of Chinese medicine and other Asian medicine is vital requirement for not only saving traditional Chinese medicine are not only important cultural resources, but also can be used as organic products to have sustainable life. Anise seed oil contains anethol, estragole, eugenol, pseudisoeugenol, methyl chavicol and anisaldehyde, coumarins, scopoleting, umbelliferon, estrols, terpene hydrocarbons, and polyacetylenes as the major compounds. The plant oil has both pharmacological and clinical effects. The pharmacological effects consist of antimicrobial, hepatopreotective, anticonvulsant, anti-inflammatory, antispasmodic, bronchodilator, estrogenic, expectorant, and insecticidal effects, and clinical effects such as nausea, constipation, menopausal period, virus, diabetes, obesity, and sedative action.

Competing Interests

The authors declare no competing interests.

1. Introduction

1.1. Anise occurrence and cultivation

Traditional medicinal herbs have been used for both food and medicinal purposes, which have obvious role in maintaining human health and improving human life quality for thousands of years (Ogbaji, Li, Xue, Shahrajabian, & Egrinya, 2018; Shahrajabian, Sun, & Qi, 2018, 2019a, 2019b, 2019c; Soleymai & Shahrajabian, 2018; Soleymani & Shahrajabian, 2012a, 2012b; Soleymani, Shahrajabian, & Naranjani, 2013). China has important potential to produce aromatic and medical plants and herbs due to its various biological diversity and different climatic conditions (Chen et al., 2013; Ogbaji, Shahrajabian, & Xue, 2013; Shahrajabian, Soleymani, Ogbaji, & Xue, 2017; Yong et al., 2017). Aromatic plants such as anise seed have a long traditional use in both folk and conventional medicine and of course in the pharmaceutical industry (Abouzid & Mohamed, 2011; Ibrahim, Mattar, Abdel-Khalek, & Azzam, 2017; Sevindik, Murathan, Yamaner, & Ayvaz, 2016; Shahrajabian, Khoshkharam, Zandi, Sun, & Qi, 2019e, Shahrajabian, Sun, & Cheng, 2019f, Shahrajabian, Sun, & Qi, 2019d). Anise (Pimpinella anisum) is a herbaceous annual plant, native to Mediterranean region and primarily grown for both fruits and seeds (Zand, Darzi, & Haj Seyed Hadi, 2012). It has been reported, it is also indigenous to Iran, India, and Turkey (Kucukkurt et al., 2009). Its fruits, known also as aniseed, were used as traditional medicine in China as early as in the 5^th century. Fruits of this plant contain fatty oil, proteins, carbohydrates, and cellulose fibers. In European countries, consumption of anise fruits is more than its production so the amount of imported anise fruits reached about 2,000 t in 2004 (Ullah, Mahmmod, & Honermeier, 2014). Among other countries Germany remains the largest spice importer of anise (Rapisarda, 2004). Sirisha and Sujathamma (2018) noticed that anise usually grows on dry rocky places, rocky crevices, fields, meadows, mountains pastures, and grasslands, and its seed germination in nature is very poor.

1.2. Anise classification and variation species

The Pimpinella anisum has common names in different countries such as: Anis vert (France); Anise seed (Japan); Anise and Star anise (the USA); Annesella (Italy); Anisa, Badian, Kuppi, Muhuri, Saunf and Sop (Iran and India); Boucage anis, Petit anise (North Africa), and anise (England) (Khare, 2007; Ross, 2001). Information on scientific names, common names, uses, origins, cultivation potential toxic compounds, and toxicity is shown in Table 1.

Table 1. Plant spices known as anise

Scientific names and synonyms	Common names	Uses	Origin/Introduction to Europe	Cultivation	Potentially toxic compounds	Toxicity
Pimpinella anisum Linne (Umbeliferae family)	Anise, anis vert, aniseed	Carminative, antispasmodic, expectorant, insecticide, bactericide, aromatic; manufacture of preserves, liquirs, and confectionery	Middle East/introduced by the Moors	Mediterranean	Anethole	Relatively safe
Illicium verum Hooker filius; Illicium anisatum Lour (Magnoliaceae family)	Star anise, Chinese star anise, badiane	Antioxidant, antimicrobial, expectorant, analgesic, and sedative	Asia/17^th century	Southeast Asia	Anethole, estragole, veranisatin A, B, and C	Toxic at high doses
Illicium anisatum Linne, Illicium japonicum, Illicium religiosum	Japanese star anise, shikimmi, skimmi	No clear medicinal effects	Asia	Sotheast Asia	Anethole Anisatins	Highly toxic (not authorized use)

1.3. Anise nutritional composition and chemical constituents

Anethole which is used in pharmaceutical, food, perfumery, and flavoring industry, is the most important constituent of anise (Ozkan & Chalchat, 2006; Tuncturk & Yildirim, 2006). The yield and anethole content of aniseed are affected by the genotype, the ecological conditions, and especially by agricultural practices such as the irrigation, plant population, fertilizer, and planting date (Acimovic et al., 2014; Asadi-kavan, Ghorbanli, Pessaraki, & Sateei, 2009; Ozel, 2009). Apart from anethole, anise is well known for essential oil, which gives it the characteristics odor and aroma. Although, the major component of anise oil is trans-anethole (75–90%), other constituents include coumarins (umbelliferone, umbelliprenine, bergapten, and scopoletin), lipids (fatty acids, beta-armyrin, stigmasterol, and its salts), flavonoids (flavonol, flavone, glycosides, rutin, isoorientin, and isovitexin), protein, and carbohydrate (Picon et al., 2010; Yamini, Bahramifar, Sefidkon, Saharkhiz, & Salamifar, 2008). It has been reported that apiaceae seed are used in food industry such as bread, biscuits, and cookies as ingredients, and in meat industry, Apiaceae are favorable spices. (Acimovic, Kostadinovic, Popovic, & Dojcinovic, 2015). Components of anise essential oil analyzed by GC-MS are shown in Tables 2 and 3. Anise methanolic extract and molecular formula is shown in Table 4. Chemical composition (%) of oleoresins of anise is shown in Table 5.

Table 2. Mean values or components of anise essential oil analyzed by GC-MS (Ullah, Mahmood, & Honermeier, 2013)

No.	Components	Kovats retention index	%
1	Estragol	1,197	0.33
2	Cis-Anethole	1,252	0.14
3	Trans-Anethole	1,287	82.1
4	δ-Elemene	1,333	0.45
5	β-Elemene	1,388	0.08
6	α-Himachalene	1,449	0.71
7	γ-Himachalene	1,478	7.0
8	α-Amorphane	1,482	0.15
9	(E)-Methylisoeugenol	1,489	0.14
10	α-Zingiberene	1,493	0.77
11	β-Himachalene	1,499	0.44
12	α-Muurolene	1,502	0.15
13	β-Bisabolene	1,506	0.38
14	β-Sesquipehllandrene	1,522	0.05
15	Spathulenol	1,580	0.04
16	Unknown	1,629	0.05
17	α-Cadinol	1,651	0.08
18	Unkown	1,831	5.95
19	Unknown	1,886	0.92

No. of identified compounds.

Table 3. Chemical composition of Pimpinella anisum essential oil analyzed by GC/MS (Singh, Kappor, Singh, de Heluani, & Caralan, 2008)

Compound	%MS	RI#	Identification Φ
α-pinene	0.1	927	MS,RI,co-GC
Sabinene	t	964	MS,RI,co-GC
Myrcene	t	983	MS,RI,co-GC
Αα-phellandrene	t	998	MS,RI,co-GC
p-cymene	0.1	1018	MS,RI,co-GC
Limonene	0.8	1023	MS,RI,co-GC
1,8-cineole	0.1	1026	MS,RI,co-GC
Cis-β-ocimene	t	1029	MS,RI,co-GC
Fenchone	5.0	1083	MS,RI,co-GC
Camphor	0.2	1138	MS,RI,co-GC
Methyl chavicol	2.3	1192	MS,RI
Endo-fenchyl acetate	0.1	1224	MS,RI,co-GC
Cis-anethole	0.5	1247	MS,RI
p-anisaldehyde	0.5	1253	MS,RI
Trans-anethole	90.1	1294	MS,RI
Total	99.9%

Trace ˂0.05; # the retention index (RI) was calculated using a homologous series of n-alkanes C₈-C₁₈;Φ_co-GC: coinjection with an authentic sample. Percentages are the mean of three runs and were obtained from electronic integration measurements using selective mass detector.

Table 4. Anise methanolic extract and molecular formula

No.	Compound name	Molecular formula
1	Anethole	C10H12O
2	Varidiflorene	C15H24
3	Eicosane	C20H42
4	Docosane	C22H46
5	Nonadecane	C19H40
6	Pentadecane	C15H32
7	Butanoic acid	C15H20O3
8	Heneicosane	C21H44
9	Octacosane	C28H58
10	Hexadecane	C20H42
11	Cyclohexane	C26H50

Table 5. Chemical composition (%) of oleoresins (in various solvents) of Pimpinella anisum analyzed by GC/MS (Singh et al., 2008)

Compounds	PS2	PS3	PS4	PS5	RI#	Identification Φ
p-cymene	—	—	t	—	1028	MS,RI,co-GC
Limonene	t	—	0.5	t	1031	MS,RI,co-GC
1,8-cineole	—	—	t	—	1034	MS,RI,co-GC
Cis-β-ocimene	—	—	t	—	1039	MS,RI,co-GC
Fenchone	1.2	0.3	0.9	0.4	1093	MS,RI,co-GC
Undecane	t	—	—	0.3	1100	MS,RI,co-GC
Camphor	t	t	t	—	1149	MS,RI,co-GC
Methyl chavicol	t	—	0.5	t	1201	MS,RI
Endo-fenchyl acetate	t	t	t	t	1226	MS,RI
p-anisaldehyde	t	—	t	t	1258	MS,RI
Trans-anethole	14.1	5.7	19.7	6.4	1295	MS,RI
Palmitic acid, methyl ester	t	0.3	t	t	1914	MS,RI
Plamitic acid	8.5	7.9	5.3	5.7	1968	MS,RI,co-GC
Palmitic acid, ethyl ester	1.2	—	—	—	1981	MS,RI,co-GC
Linoleic acid, methyl ester	t	0.7	0.1	t	2080	MS,RI,co-GC
Oleic acid methyl ester	t	2.8	0.4	0.5	2086	MS,RI,co-GC
Oleic acid	57.9	75.5	57.5	63.5	2130	MS,RI,co-GC
Oleic acid, ethyl ester	11.0	—	—	—	2146	MS,RI,co-GC
Stearic acid	1.9	—	—	—	2157	MS,RI,co-GC
2-oleoylglycerol	2.3	2.8	2.1	3.6	—	MS
Squalene	—	t	0.2	t	—	MS
Octacosanal	t	t	0.4	0.6	—	MS
Stigmasterol	t	t	0.4	0.5	—	MS

PS2: ethanol extract; PS3: methanol extract; PS4: -hexane extract; PS5: petroleum benzene extract. Trace: ˂0.05; # the retention index (RI) was calculated using a homologous series of n-alkanes C₆-C₂₂; Φ coinjection with an authentic sample. Percentages are the mean of three runs and were obtained from electronic integration measurements using a selective mass detector.

The oil content and fatty acid composition of anise is shown in Table 6. Composition of Pimpinella anisum essential oil (%) of various origins has been shown in Table 7. Properties of aniseed oil Are shown in Table 8. Total phenols, carotenoids, and tannins in ethanolic extract of anise are shown in Table 9. HPLC-analysis of polyphenolic compounds in ethanolic extracts of anise is shown in Table 10. Oil yield and fatty acid composition of Tunisian and Egyptian anise seeds (Table 11).

Table 6. Oil content and fatty acid composition of anise (Alfekaiki, 2018)

Element	Mineral contents (%)
Carbon	34.5
Oxygen	21.1
Nitrogen	ND
Aluminum	20.5
Bromine	14
Iron	ND
Copper	3.63
Lanthanum	5.85
Palladium	0.35
Sulfur	0.15
Potassium	ND
Magnesium	ND

Table 7. Composition of Pimpinella anisum essential oil (%) of various origins (Saibi, Belhadj, & Benyoussef, 2013)

Components	Algeria		Reported compositions
		Turkey	Turkey	Portugal
Linalool	0.3	-	0.8	-
Terpinene 4-ol	-	-	0.6	-
Methyl chavicol	-	-	0.8	-
α-terpineol	-	-	1.0	-
Estragol	1.9	0.6	-	2.2
Anisaldehyde	-	0.9	0.5	1.9
Cis-anethole	0.5	0.3	0.1	Tr
Trans-anethole	92.4	95.4	89.5	92.5
Methy eugenol	-	-	0.6	-
γ-himachalene	1.1	0.8	-	-
Zingiberene	0.3	-	-	-
Anesic acid	-	0.5	-	-
Anisylacetone	0.3	-	0.2	-
Anisyl alcohol	-	-	0.1	-
o-isoeugenol	1.9	-	0.2	-
Trans-Pseudoisoeugenyl-2-methybutyrate	-	-	-	0.1
Butanoic acid, 2-methyl-,4-methody-2(3-methyloxiranyl)phenyl ester	0.3	-	-	-

Table 8. Properties of aniseed oil (Yadav, Mahadwad, Kshirsagar, & Gite, 2015)

Sr. no.	Parameter	Results
1	Color	Pale yellow
2	Specific gravity	0.987
3	Saponification value	168.3
4	Acid value	2.55
5	Iodine value	99
6	Refractive index	1.55
7	Odor	Sweet like Anethole

Table 9. Total phenols, carotenoids, and tannins in ethanolic extract of anise (Tolba, El-Sherif, & El-Sherif, 2012)

Total phenols (as mg gallic acid)	64.63
Carotenoids (mg/100g)	23.33
Tannins (mg as tannic acid)	83.31

Table 10. HPLC-analysis of polyphenolic compounds in ethanolic extracts of anise

Compound		Anise
	Rt	Area (%)	ppm
Protocatchoi	-	-	-
Catechein	2.467	1.2923	3.310
Pyrogallo	2.494	1.4354	6.391
Chlorogenic	2.785	4.0396	2.875
Caffeic	-	-	-
Vanillic	-	-	-
Syrinic	3.933	1.0083	0.137
Caffeine	-	-	-
P. coumaric	6.090	0.6257	0.060
Ferullic	-	-	-
Salycilic	6.541	2.3829	1.704
Coumarin	6.679	1.3413	0.132
Cinnamic	8.447	0.2376	0.017
Chrisin	11.708	0.2359	1.285
Hypersoid	4.936	3.9599	47.103
Quecettin	7.384	1.2037	17.239
Luteolin	7.756	1.8574	159.300
Kampferol	8.757	0.5232	7.177
Apegnin	8.860	0.3272	6.880
Total area of phenolic compounds		7360.32203
Total area of flavonoid compounds		7681.33273

Table 11. Oil yield and fatty acid composition (%) of Tunisian and Egyptian anise (Pimpinella anisum) seeds (means of six replicates ± S.D). Values with different superscripts (a-b) are significantly different at p ˂ 0.05 (Rebey et al., 2017)

	TAS	EAS
Oil yield (%)	11.60 ± 0.03^a	9.82 ± 0.02^b
Saturated fatty acid (SFA)(%)
Capric acid (C10:0)	0.16 ± 0.03^a	0.11 ± 0.01^a
Lauric acid (C12:0)	0.52 ± 0.01^a	0.44 ± 0.02^a
Myristic acid (C14:0)	0.07 ± 0.01^a	0.02 ± 0.00^a
Palmitic acid (C16:0)	4.90 ± 0.22^b	13.20 ± 0.09^a
Stearic acid (C18:0)	0.85 ± 0.04^a	0.66 ± 0.01^a
Arachidic acid (C20:0)	0.07 ± 0.01^a	0.07 ± 0.00^a
Total	6.57	14.5
Unsaturated fatty acid (UFA)(%)
Petroselinic acid (C18:1 ∆6)	46.60 ± 0.22^a	38.40 ± 0.11^b
Oleic acid (C18:1 ∆9)	21.05 ± 0.08^a	18.47 ± 0.13^b
Lioleic acid (C18:2)	22.99 ± 0.44^a	23.18 ± 0.22^a
Linolenic acid (C18:3)	1.07 ± 0.01^a	0.58 ± 0.04^b
Total	91.71	80.63

Notes: Values with different superscripts (a-b) are significantly different at p ˂ 0.05 (means of six replicates); SFA: saturated fatty acid; UFA: unsaturated fatty acid.

1.4. Potential health benefits, medicinal uses of anise in modern medicine industry

Seeds of anise are used as analgesic in migraine and also as carminative, aromatic, disinfectant and diuretic in traditional medicine (Amin, 2005). In some traditional texts, anise is mentioned for melancholy, nightmare, and also in treatment of epilepsy and seizure (Mirheydar, 2001). Shobha, Rajeshwari, and Andallu (2013) noticed that aniseed is a potent antiperoxidative and antidiabetic agent and thereby, possess a vast spectrum of applications and exploitations in the food and drug industry. Cifti, Guler, Dalkilic, and Nihat Ertas (2005) showed that anise oil has anethole as active ingredients and also eugenol, methylchavicol, anisaldehyde, and estragole. Rebey et al. (2017) revealed that aniseeds might constitute a novel source of natural antioxidants and could be used as food additive. Acimovic et al. (2014) found that drought cause a significant decrease in thousand seed weight, germination energy, and total germination as well as essential oil content in anise. Contrary to this finding, the content of trans-anethole was significantly higher in the dry year. Ibrahim (2008) reported that according to the traditional thinking, drinking anise by boys maybe harmful to their reproductive system. Kreyidyyeh, Usta, Knio, Markossian, and Dagher (2003) found that extracts of the aniseeds are used as medicine for their diuretic and laxative effect, expectorant and antispasmodic action, and their ability to ease gastric pain and flatulence. Ibrahim et al. (2017) reported that waste residues of anise and star anise are promising new sources of phenolic antimicrobial compounds, which offer new commercial opportunities to pharmaceutical industry. They have suggested that combination of anise waste extracts with some antibiotics leads to new choice for treatment of infectious diseases and waste extracts may act as activity modifying agent for antibiotics. Islam, Khan, Rakhshanda, Mahdi, and Chowdhury (2016) reported that aniseed extracts showed positive antibacterial effects on only three bacteria, named, Bacillus cereus, Bacillus subtilis, and Streptococcus pneumonae. They have also found that the phytochemical analysis of the aqueous extract revealed the presence of alkaloids, flavonoids, saponins, tannins, terpenoids, phenolic compounds, and cardiac glycosides. Fouda, Elewady, Shalabi, and Habouba (2014) indicated that the anise extract shows good performance as corrosion inhibitor 1 M HCl, and also the anise extract inhibits the corrosion by getting adsorbed on the etal surface following Langmuir adsorption isotherm. Positive antimicrobial effects produced by methanol, ethanol, and aqueous extract of aniseed are shown in Table 12. Antifungal activity of the essential oils, which expressed through the minimal inhibitory concentrations, is shown in Table 13. Positive phytochemical assay of aniseed in aqueous extract is shown in Table 14. Antifungal activity of anise fluid extract and essential oil by the diffusion method is shown in Table 15. Antifungal activity of anise fluid extract and essential oil by the dilution method is shown in Table 16.

Table 12. Positive antimicrobial effects (average zone of inhibition) produced by methanol, ethanol, and aqueous extract of aniseed, and that in positive controls (Islam et al., 2016)

Extraction condition	Day of extraction		Zone of inhibition (mm)
		Bacillus cereus	Bacillus subtilis	Streptococcus pneumonae
Methanol extraction	2^nd day	19.03 ± 1.53	18.77 ± 0.55	12.07 ± 1.39
	5^th day	20.13 ± 0.32	19.27 ± 1.2	14.5 ± 0.65
	7^th day	18.43 ± 1.15	15.63 ± 1.16	14.87 ± 0.96
	Positive controls	28.61 ± 0.91	23.44 ± 0.5	31.67 ± .88
Ethanol extraction	2^nd day	14.90 ± 0.36	14.01 ± 0.7	12.17 ± 0.12
	5^th day	14.52 ± 0.95	12.19 ± 0.9	13.00 ± 0.7
	7^th day	14.78 ± 0.94	16.08 ± 1.36	14.44 ± 1.17
	Positive controls	28.20 ± 1.3	23.80 ± 1.47	31.39 ± 1.67
Aqueous extraction	2^nd day	11.05 ± 0.69	10.83 ± 1.18	-
	Positive controls	29.56 ± 0.91	24.83 ± 1	-

The results represent the mean ± SD of values obtained from three independent experiments.

Table 13. Antifungal activity of the essential oils is expressed through the minimal inhibitory concentrations (mg/ml) (Starovic et al., 2016)

Fungal species	Anise
Bipolaris/Drechslera sorociniana	0.325
Fusarium subglutinans	0.0775
Fusarium verticillioides	0.325
Fusarium oxysporum	0.55
Fusarium tricinctum	0.325
F. sporotrichioides	0.055
F. equiseti	5.5
F. incarnatum	7.75
Macrophomina phaseolina	0.325
F. proliferatum	7.75

*Values of MIC, followed by the same letter are not significantly different (p ˂ 0.05) according to Duncan^,s multiple range test.

Table 14. Positive phytochemical assay of aniseed in aqueous extract (Islam et al., 2016)

S1. No	Test	Interference	Results
1	Tannin	Bluish-black or brownish-green precipitate	Present
2	Saponin	Frothing	Present
3	Terpenoid	Reddish brown precipitate	Present
4	Flavonoid	Appearance of yellow color	Present
5	Cardiac glycoside	Appearance greenish and violet ring	Present
6	Alkaloids	Creamy-white precipitate	Present
7	Phenolics	Appearance of dark green color	Present
8	Steroid	Appearance of red colored product	Present

Ghazanfarpour, Sadeghi, Abdolahian, and Roudsari (2016) showed that anise can alleviate the side effects of hot flashes, so they suggest usage of herbal medicine can be seen as an alternative treatment for women experiencing hot flashes.

Table 15. Antifungal activity of anise fluid extract and essential oil by the diffusion method (Kosalec, Pepeljnjak, & Kustrak, 2005)

	Fungi		Inhibition zone (mm)
		Reference substance^a	Fluid extract	Essential oil
Yeasts	Candida albicans	18	12	29
	C. tropicalis	23	11	AE^c
	C. pseudotropicalis	18	10	AE^c
	C. parapsilosis	18	11	30
	C. krusei	17	11	AE^c
	C. glabrata	18	0	21
	Geotrichum spp.	19	GPA^b	17
Dermatophytes	Trichophyton rubrum	21	20	25
	T. mentagrophytes	24	29	23
	Microsporum canis	21	26	27
	M. gypseum	21	23	21

^a Nistatin as reference substance (0.5 mg mL⁻¹) for yeasts and ketoconazole for dermatophytes.

^b GPA, growth promotion activity.

^C AE, fungicidal influence of aerosol (0.7 µL cm⁻³).

Table 16. Antifungal activity of anise fluid extract and essential oil by the dilution method (Kosalec et al., 2005)

	Fungi			Antifungal activity
		Reference substance^a (µg mL^−1)		Fluid extract (%, V/V)		Essential oil (%, V/V)
		MIC^b	MFC^c	MIC	MFC	MIC	MFC
Yeasts	Candida albicans	0.5	1	19	25	1.00^d	2.00^d
	C. parapsilosis	1	2	18	25	0.10^d	0.20^d
	C. tropicalis	1	2	18	25	0.10^d	0.20^d
	C. pseudotropicalis	0.5	1	17	20	0.20^d	0.39^d
	C. krusei	0.5	1	20	25	0.20^d	0.39^d
	C. glabrata	2	4	nt	nt	0.10^d	0.20^d
	Geotrichum spp.	2	4	nt	nt	1.56^d	3.12^d
Dermatophytes	Trichophyton rubrum	0.25	0.5	1.5	3	0.20^d	0.39^d
	T. mentagrophytes	0.50	1	9.0	20	0.78^d	1.56^d
	Microsporum canis	0.25	0.5	2.5	5	0.10^d	0.20^d
	M. gypseum	0.25	0.5	7.0	15	0.20^d	0.39^d

^a Nistatin as reference antimycotic against yeasts tested and ketoconazole against dermatophytes tested.

^b MIC, minimum inhibitory concentration.

^c MFC, minimum fungicidal concentration.

^d Significantly different from fluid extract (p˂0.01).

Nt, not tested.

Qualitative analysis of methanolic extract and various fractions is shown in Table 17. Phytoceuticals in methanolic extract and various fractions of aniseed is shown in Table 18. Essential oil composition of Tunisian and Egyptian anise seeds is shown in Table 19. Antimicrobial activity of anise EO in disc-diffusion method is shown in Table 20. The major molecular compounds identified in the essential oils of cumin, clove, cinnamon, anis, and laurel using gas chromatography mass spectrometry (GC-MS) is shown in Table 21. Minimum inhibitory concentration (MIC) of anise EO is shown in Table 22. Antioxidant activity of aniseed extracts as affected by maturity stages is shown in Table 23. GC-MS chromatography of anise oil components is shown in Table 24.

Table 17. Qualitative analysis of methanolic extract and various fractions (Shobha & Andallu, 2016)

Test	Me	He	Be	Ea	n-But	Aq
Carbohydrates
a. Molisch test	+	-	-	+	-	++
b. Fehling^,s test	-	-	-	-	-	+
c. Barfoed^,s test	-	-	-	-	-	-
d. Benedict^,s test	-	-	-	-	-	+
Amino acids
a. Ninhydrin test	+	-	-	+	-	++
b. Hopkin^,s-Cole test	-	-	-	-	-	-
c. Erhlich^,s test	-	-	-	-	-	-
d. Pauly^,s test	-	-	-	-	-	-
e. Nitroprusside test	-	-	-	-	-	-
Flavonoids
a. Sodium hydroxide test	++	+	+	+++	+	+
b. Sodium acetate test	++	+	+	+++	+	+
c. Sulfuric acid test	++	+	+	+++	+	+
Polyphenols and tannins
a. Ferric chloride test	++	+	+	+++	+	+
b. Potassium dichromate test	++	+		+++	+	+
c. Potassium ferricyanide test	++	+		+++	+	+
Sappnins
a. Foam test	+	++		++	+	+
Sterols
a. Leibermann Buchard^,s test	+	+++		+	+	+
b. Salkowski test	+	++			-	+

Me, Methanolic extract; He, Hexane; Be, Benzene; Ea, ethyl acetate; n-But, n-Butanol; Aq, Aqueous

Table 18. Phytoceuticals in methanolic extract and various fractions of aniseed (Shobha & Andallu, 2016)

Sample extract	Total phenolic (mg/100 g) GAE	Total flavonoids (mg/100 g) RE	Total Flavonols (mg/100 g) RE
Methanol	0.63 ± 2.3	0.32 ± 0.9	0.47 ± 1.9
Hexane	0.16 ± 2.9	0.20 ± 0.5	0.24 ± 0.5
Benzene	0.23 ± 0.6	0.27 ± 1.4	0.34 ± 0.3
Ethyl acetate	0.77 ± 0.3	0.45 ± 1.6	0.55 ± 0.1
n-butanol	0.42 ± 1.7	0.15 ± 1.1	0.41 ± 0.9
Aqueous	0.15 ± 2.1	0.07 ± 1.7	0.08 ± 1.4

Values are mean ± SEM of three replicates.

Table 19. Essential oil composition of Tunisian and Egyptian anise (Pimpinella anisum) seeds (means of six replicates ± S.D). Values with different superscripts (a-b) are significantly different at p ˂ 0.05 (Rebey et al., 2017)

			%	%
Compunds*	RI^a	RI^b	TAS	EAS
Terpene hydrocarbons			0.13	0.04
Linalool	1097	1557	0.13 ± 0.01^a	0.04 ± 0.04^b
Oxygenated monoterpene			0.06	0.02
α-Terpinene	1018	1249	0.06 ± 0.01^a	0.02 ± 0.00^a
Phenylpropanoids			95.78	94.99
Anisole	918	1720	0.97 ± 0.05^a	0.52 ± 0.03^b
Estragole	1197	1430	0.20 ± 0.03^b	3.74 ± 0.13^a
Trans-anethole	1253	1740	94.30 ± 0.01^a	90.41 ± 0.22^b
p-Anisaldehyde	1250	1718	0.17 ± 0.01^a	0.10 ± 0.07^a
Cis-isoeugenol	1359	2180	0.14 ± 0.01^a	0.22 ± 0.02^a
Sesquiterpene hydrocarbons			3.95	2.48
β-Elemene	1388	1465	0.07 ± 0.66^a	0.09 ± 2.11^a
γ-Himachalene	1484	1690	2.32 ± 0.04^a	1.08 ± 0.01^b
Zingiberene	1494	1672	0.30 ± 0.03^a	0.25 ± 0.03^a
β-Himachalene	1505	1942	0.12 ± 0.02^a	0.11 ± 0.01^a
β-Bisabolene	1506	1832	0.19 ± 0.02^b	0.85 ± 0.01^a
Isolongifolene	1532	2003	0.04 ± 0.00^a	0.02 ± 0.00^a
Diepi-α-cedrene	1575	2020	0.91 ± 0.02^a	0.08 ± 0.00^b
Total identified			99.74	97.53

Notes: Volatile compounds percentages in the same line with different letters (a-b) are significantly different at p ˂ 0.05 (means of six replicates). RI^a order of elution in apolar column (HP-5); RI^b order of elution in polar column (HO Innowax), MS: mass spectrum.

Table 20. Antimicrobial activity of P. anisum EO in disc-diffusion method (30 µl/disc) (Abdel-Reheem & Oraby, 2015)

Microorganism		Inhibition zones in mm
	P. anisum oil	Corn oil	Streptomycin
Staphylococcus aureus	15	0	19
Streptococcus pyogenes	12	0	17
Escherichia coli	15	2	19
Salmonella typhi	17	1	20
Pseudomonas aeruginosa	10	0	15
Enterococcus faecalis	16	0	18
Micrococcus luteus	14	3	20
Candida albicans	10	1	ND

ND= not detected.

Table 21. The major molecular compounds identified in the essential oils of cumin, clove, cinnamon, anis, and laurel using gas chromatography mass spectrometry (GC-MS) (Sergio, Fabiola, Guadalupe, Blanca, & Leon, 2013)

			Essential oils
Compound	Cumin (Cuminum cyminum)	Clove (Eugenia cyryophyllata)	Cinnamon (Cinnamomum verum)	Anis (Pimpinella anisum)	Laurel (Laurus nobilis)
α-Pinene	×		×
Limonene	×			×	×
γ-Terpinene	×
Cuminaldehyde	×
Alcohol cuminic	×
Eugenol		×
Caryophyllene		×	×
Acetate of eugenyle		×
Chavicol		×
Carene			×	×
Cinnamaldehyde			×
Geraniol			×
β-Myrcene					×
Germancrene				×
Eucalyptol					×
Ocimene					×
Trans-anethol				×
Cis-anethol				×

Table 22. Minimum inhibitory concentration (MIC) of Pimpinella anisum EO (Abdel-Reheem & Oraby, 2015)

Microorganism		Inhibition zones in mm
	P. anisum oil	Corn oil	Streptomycin
Staphylococcus aureus	3.0	N	1.5
Streptococcus pyogenes	4.0	N	1.0
Escherichia coli	2.5	10.0	1.5
Salmonella typhi	2.0	N	1.5
Pseudomonas aeruginosa	3.0	N	1.5
Enterococcus faecalis	2.0	N	1.0
Micrococcus luteus	2.0	9.0	1.5
Candida albicans	4.0	N	ND

ND = not detected, N = bacteria not effected at any concentration.

Table 23. Antioxidant activity of aniseed (Pimpinella anisum L.) extracts as affected by maturity stages (Rebey et al., 2019)

			Antioxidant assays
		DPPH (IC50 µg/mL)	Chelating ability (IC50 mg/mL)	Reducing power (EC50 µg/mL)
Turkey	Immature	103.45	108.8	1234.28
	Intermediate	42.97	57.81	947.05
	Mature	18.97	12.46	684.73
Serbia	Immature	153.18	122.76	982.27
	Intermediate	58.04	68.09	687.41
	Mature	20.18	10.89	565.11
Egypt	Immature	103.02	130.55	1080.39
	Intermediate	62.17	68.51	764.22
	Mature	16.77	13.17	618.82
Tunisia	Immature	96.21	97.34	1154.33
	Intermediate	35.27	52.84	755.88
	Mature	12.87	9.23	523.47

Table 24. GC-MS chromatography of anise oil components (Obaid, Al Janabi, & Taj-Aldin, 2017)

No.	Compound	Rt (min)	M. Weight	Area
1	Alfa-pinene	5.61	136	0.63
2	Estragole	13.81	148	3.7
3	Anethole	15.99	148	18.54
4	β-Himachalene	20.9	204	8.5
5	Αα-Himachalene	21.87	204	6.6
6	cyclohexene	22.84	82	2.01
7	2-Methoxy-5-(1-propenyl) phenol	31.05	164	2.93

Values are mean ± SEM of three replicates.

ND = not detected.

Alfa-pinene, estragole, anethole, β-Himachalene, α-Himachaelen, cyclohexene, and 2-Methoxy-5-(1-propenyl) phenol are shown in Figures 1–7.

Figure 1. Alfa-pinene.

Figure 2. Estragole.

Figure 3. Anethole.

Figure 4. β-Himachalene.

Figure 5. α-Himachaelen.

Figure 6. Cyclohexene.

Figure 7. 2-Methoxy-5-(1-propenyl) phenol.

Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of anise dried extract on different fungi species is shown in Tables 25 and 26, respectively. Biochemical reactions leading to phenylpropenes in plant and the reactions catalyzed by known enzymes is shown in Figure 8.

Figure 8. Biochemical reactions leading to phenylpropenes in plants. The reactions catalyzed by known enzymes are indicated (Koeduka, Baiga, Noel, & Pichersky, 2009) .

Table 25. Minimum inhibitory concentration (MIC) of P. anisum dried extract on different fungi species (Yazdani et al., 2009)

			Dried extract concentration (mg/ml)
Fungi species	64	32	16	8	Control
Aspergillus niger	-	-	-	-	-
Candida albicans	+	+	+	-	-
Microsporum canis	+	+	+	-	-
Epidermophyton fluccosum	+	+	+	-	-
Trichophyton mentagrophytes	+	+	+	-	-

+ = Positive inhibition, - = Negative inhibition

Table 26. Minimum fungicidal concentration (MFC) of P. anisum dried extract on different fungi species (Yazdani et al., 2009)

				Dried extract concentration (mg/ml)
Fungi species	256	128	64	32	16	8	4	control
Aspergillus niger	-	-	-	-	-	-	-	-
Candida albicans	+	-	-	-	-	-	-	-
Microsporum canis	+	+	-	-	-	-	-	-
Epidermophyton fluccosum	+	+	+	+	+	+	-	-
Trichophyton mentagrophytes	+	+	+	+	-	-	-	-

+ = Positive inhibition, - = Negative inhibition

Uysal, Kara, Algur, Dumlupinar, and Nuri Aydogan (2007) reported that fruits of anise use for treatment of infections, angina, bronchitis, gastritis, laryngitis, and migraine. Mohamed, Abdelgadir, and Almagboul (2015) showed that the petroleum ether, chloroform, ethyl acetate, and methanol extracts of P. anisum (1:10 and 2:10) were highly active (30–40 mm) against B. subtilis. The ethyle acetate extract exhibited moderate activity (15 mm) against Escherichia coli and low activity (13 mm) against Ps. aeruginosa. The methanol extract of P. anisum showed high activity (16 mm) against E. coli, low activity (13 mm) against Ps. aeruginosa, and the methanol extract have variable activity against all test organisms.

Yazdani et al. (2009) concluded that anise can be a candidate for further studies due to their antifungal potencies. Rebey et al. (2019) indicated that the determination of optimal periods and provenances for antioxidant accumulation can be used to evaluate the quality of aniseeds and could be important for industries. Khudor, Jasim, Malik, and Zahra (2013) mentioned that Pimpinella anisum and some antibiotics by disc diffusion methods and minimum inhibitory concentration, the results showed these bacterial isolates were sensitive to the aqueous extract compared with methanol, acetone, and petroleum ether and were more sensitive to vancomycin compared with other antibiotics. Karimzadeh et al. (2012) indicated the anticonvulsant and neuroprotective effects of anise oil, likely via inhibition of synaptic plasticity. Bagdassarian, Bagdassarian, and Atanassova (2013) stated that the seeds of anise are rich in phytochemical contents, which possessed high antioxidant and antimicrobial activities and they can be used for health supplement and pharmaceutical benefits. Aydemir, Cifci, Aviyente, and Bosgelmez-Tinaz (2018) observed that trand-anethole has the potential to inhibit QS (Quorum sensing)-regulated virulence factors in P. aeruginosa by binding to LasR protein, similar to its natural ligand N-(3-oxododecanoyl)-_L-homoserine lactone. Mahood (2012) concluded that anise oil extract can decrease signs of polycystic ovary syndrome (PCOS) in the ovarian tissue and altered concentrations of luteinizing hormone. Singh et al. (2008) stated that the antioxidant potency of anise oil and its methanol and ethanol oleoresins, can be utilized for protecting fat-containing foods. Diaz et al. (2014) stated that mixture of chamomile and star anise decrease the completion percentage of the activated carbon, delayed the appearance of diarrhea and decrease the number of evacuations in comparison with the control treatments. They have suggested the combination of chamomile and star anise as an alternative antidiarrheal treatment. Ibrahim (2008) concluded that anise oil administration inhibited GST expression, besides decreasing testosterone, T3 and T4 hormones and inhibiting sperm counts and sperm motility. Al-Omari, Qaqish, and Al-Qaoud (2018) concluded that anise possesses immunomodulatory activity when apply orally in mice and selectively activates cell-mediated immune mechanisms. Cifti et al. (2005) indicated that anise is an annual herb indigenous to Iran, India, Turkey, and many other warm regions and it could be considered as a potential natural growth promoter for poultry. Amina, Nadia, Kahloulakhaled, Nesrine, and Abdelkader (2016) showed that the aqueous extract of P. anisum L. can have a corrective effect against nephrotoxicity induced by lead, so P. anisum L. has a beneficial impact on the kidneys intoxicated with lead acetate. Mosaffa-Jahromi et al. (2017) suggested that anise oil could be a promising choice of treatment for depressed patients with irritable bowel syndrome. Shahamat, Abbasi-Maleki, and Mohammadi Motamed (2015) concluded that P. anisum possesses an antidepressant-like activity similar to that of fluoxetine, which has a potential clinical value for application in the management of depression. Felsociova et al. (2015) reported that the most hopeful antifungal activity and killing effect against all tested penicillia was found to be Pimpinella anisum and Origanum vulgare L. Rattan, Satpathy, and Gupta (2014) showed that a formulation of biscuits with anise extract was prepared for enhancing the healthy beneficial usage of biscuits and nutraceutical, in conditions like cough or sore throat with a simultaneous attainment of the other health benefits. Kargozar, Azizi, and Salari (2017) stated that Pimpinella anisum is effective in the treatments of acute menopausal syndrome with different mechanisms. Barbalho et al. (2015) suggested that P. anisum has potential to be used to control lipid levels. Radaelli et al. (2016) suggested that the use of essential oil from anise might serve as an alternative to the use of chemical preservatives in the control and inactivation of pathogens in commercially produced food systems. Kucukkurt et al. (2009) demonstrated that aniseed could be used at 30 g/kg level in quail diets an increased antioxidant activity with glutathione (GSH) and a decreased blood malondialdehyte (MDA) levels. Abdel-Reheem and Oraby (2015) found that Pimpinella anisum essential oil residuals have high inhibitory effect for Salmonella typhi, Enterococcus faecalis, Staphylococcus aureus, E. coli, and Micrococcus Iuteus. Pavela (2014) stated has the essential oil from Pimpinella anisum fruits and trans-anethole were toxic for Daphnia magna (62–92% mortality) and significantly reduced it fertility at high concentrations (35–50 µL mL⁻¹) and long exposure (48 h). However, no negative effect on Daphnia mortality or fertility was found at shorter exposure times (6 h) and lower concentrations (20 µL mL⁻¹). Womeni, Djikeng, Tiencheu, and Linder (2013) found that the powder of Pimpinella anisum has a special potent antioxidants for stabilization of crude soybean oil. Changizi-Ashtiyani et al. (2017) observed that the simultaneous use of ethanolic extract of P. anisum during gentamicin (GM) administration is recommended to reduce its nephrotoxicity effects. Barbalho et al. (2015) reported that P. anisum has potential to be used to control lipid levels. Kadan, Rayan, and Rayan (2013) noted that anise could be one of the foods that attribute to cancer prevention and treatment. It could be a natural source of novel anticancer compounds with antiproliferative and apoptotic properties. Bekara et al. (2015) observed that aniseed aqueous extract was effective in reducing the level of some of biochemical parameters and ameliorate behavior of intoxicated rats by lead. Nahidi, Kariman, Simbar, and Mojab (2012) concluded that P. anisum is effective on the frequency and severity of hot flashes in postmenopausal women. Hosseinzadeh, Tafaghodi, Abedzadeh, and Taghiabadi (2014) stated that P. anisum aqueous and ethanolic extracts can increase milk production in rats. Sharifi, Kiani, Farzaneh, and Ahmadzadeh (2008) reported that anise is commercially cultivated in Iran and has been used in medicinal applications; moreover, their oil can be effective for protection of fresh fruits facing fungi and its essential oil can be considered as a potential, broad spectrum and safe substitute of chemical agents. Ashraffodin Ghoshergir et al. (2015) showed the effectiveness of anise in relieving the symptoms of postpartum depression. Shirzadi, Abbasi-Maleki, and Zanbouri (2017) noted that P. anisum ethanolic extract is effective in suppression of morphine physical dependence and further studies are needed to find out the responsible constituents and also the exact mechanisms of actions. Niksokhan, Hedarieh, Najafifard, and Najafifard (2015) observed that 250 and 300 mg/kg of hydroalcoholic extract of P. anisum seed significantly increased the duration of open arm ledges and decreased the duration of closed arm ledges in maze; in addition, this treatment resolved anxiety and relieved anxiety in rats. Nikfarjam, Bahmani, and Heidari-Soureshjani (2016) decribed that Pimpinella anisum is one the most important native medicinal plants of Iran with antianxiety properties. Amini, Tajabadi, Khani, Labbafi, and Tavakoli (2018) identified components of the essential oils and obtained results showed that Pimpinella anisum L. showed the most fumigant toxicity on the storage pests. The pharmacological effect of anise is shown in Table 27.

Table 27. The pharmacological effects of Pimpinella anisum (Shojaii & Abdollahi Fard, 2012)

System	Effect
Organism	Antibacterial
	Antifungal
	Insecticidal
	Antiviral
Muscle	Muscle relaxant of tracheal chain
	Antispasmodic and relaxant of anococygeus smooth muscle
Nervous system	Anticonvulsant
	Analgesic
	Conditioned place aversion in morphine dependence
Gastrointestinal	Laxative
	Increase glucose absorption from the jejunum
Renal	Reduce volume of urine by increase activity of the renal Na^+-K^+-ATPase
Endocrine	Antidiabetic
	Hypolipidemic
Immune system	Antioxidant
Others	Reduction of menopausal hot flashed
	Growth promoter of day-old broilers
	Reduction of pain in dysmenorrhea

2. Conclusion

The followings conclusion could be drawn for the discussions. Pimpinella asnisum L. is an aromatic, annual grassy herb with 30–50 cm height, white flowers and small green to yellow seeds with secondary feather like leaflets of bright green, twice pinnate. Due to all positive characteristics, such as antidiabetic, hypolipidemic, antioxidant activities, anticancer, and antimicrobial properties, both seeds and essential oils of anise is promising for safe use as super food supplements and raw constituents in the both pharmaceutical and food industries. Keeping the traditional knowledge of Chinese medicine and other Asian medicine is vital requirement for not only saving traditional Chinese medicine as the important cultural resources, but also using organic products to have sustainable life. Anise seed oil contains anethol, estragole, eugenol, pseudisoeugenol, methyl chavicol and anisaldehyde, coumarins, scopoleting, umbelliferon, estrols, terpene hydrocarbons, and polyacetylenes as the major compounds. The plant oil has both pharmacological and clinical effects. The pharmacological effects consist of antimicrobial, hepatopreotective, anticonvulsant, anti-inflammatory, antispasmodic, bronchodilator, estrogenic, expectorant and insecticidal effects, and clinical effects such as nausea, constipation, menopausal period, virus, diabetes, obesity, and sedative action. More clinical studies are necessary to uncover the numerous substances and their effects in ginseng that contribute to public health.

Additional information

Funding

The authors received no direct funding for this research.

Notes on contributors

Wenli Sun

Dr. Wenli Sun is an assistant researcher working on related topics of allelopathic characteristics of medicinal plants, especially traditional Asian herbs, fruits and crops. She is also working on topics related to sustainable agriculture and farming. Her full profile is available in http://orcide.org/0000-0002-1705-2996

Mohamad Hesam Shahrajabian

Dr. Mohamad Hesam Shahrajabian is a senior researcher of Agronomy and Biotechnology. His full profile is available in https://orcide.org/0000-0002-8638-1312

Qi Cheng

Prof. Dr. Qi Cheng, is a professor of biotechnology and his researches have connected with agrobiotechnology and agrotechnology. His full profile is available in http://orcide.org.0000-0003-1269-6386. All these scholars are researchers in Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, and Nitrogen Fixation Laboratory, Qi Institute, Building C4, No. 555 Chuangye Road, Jiaxing 314000, Zhejiang, China (Corresponding author).