Phytochemical Analysis of Ethanolic Extracts of Three Sudanese Tribulus species

Napata Scientific Journal, Vol.1 (1) 2022 pp. 30-44

Original

Phytochemical Analysis of Ethanolic Extracts of Three Sudanese Tribulus species

  1. A. Hilmi (1,2), H. H. EL-Kamali (3)* , A. M. Aldai (3)

1 Department of Microbiology & Parasitology, Medicine Program, Napata College,  Khartoum State, Sudan . 2Department of Biochemistry and Molecular Biology, Faculty of Science, Gezira University,  Wad Medani, Gezira State, Sudan.  3Department of Botany, Faculty of Science and Technology, Omdurman Islamic University, Khartoum State, Sudan

Corresponding Author: zahir.hilmi@gmai.com

Abstract:

This   study aims to explore the phytochemically bioactive compounds of three Sudanese Tribulus species. Threewild species of Tribulus; T. terrestris, T.longipetalous and T.pentandrus,were collected from KhartoumState.  Their bioactive compounds were extracted by ethanol, from their roots, aerial parts, and fruits.  Gas-Chromatography-Mass Spectroscopy (GC-MS) was used for phytochemical analysis of their bioactive compounds.In T. terrestris different compounds (20 from roots), (27 from aerial parts) & (29 from fruits) were identified by GC/MS. Six classes of  chemical compounds were found; sugars, fatty acids, terpenes, steroids, alkanes and alcohols. Major components were sucrose (28.72%) in roots and 4-O-methyl-mannose in aerial parts (49.60%) and in fruits (43.66%).

In T.longipetalous many compounds were identified; 23 from roots, 20 from aerial parts, and 29 from fruits. These compounds were classified into four groups; Sugars, fatty acids, terpenes alcohols.  The most commonly detected compound was 4-O-methyl mannose; 73.11% (in roots), 43.24% (in aerial parts) and 35.75% (in fruits). In T.pentandrus more compounds were identified, 31 (roots), 31 (aerial parts) and 42 from fruits. These compounds were grouped into eight classes; sugars, fatty acids, terpenes, steroids, alkanes, alcohols, steroidal sapogenins and alkaloids. The major component also was 4-O-methylmannose 47.86 % (in roots), 44.75 % (in aerial parts) and 55.05 % (in fruits).The most commonly found classes of compounds in the three Tribulus species were; sugars, fatty acids, terpenes, steroids and alcohols.

The major compound found in all parts of the three Tribulus species was 4-O-methylmannose. In addition other compounds were identified from T.pentandrus

Keywords:Tribulus sp, phytochemical analysis, ethanolic extracts, sucrose, 4-O-methylmannose, gamolenic acid, alkaloids, saponins.

 

 

 

Introduction:

 Tribulus species are a group of annual herbs belonging to theZygophyllaceae family with 25 genera and 250 species.  It is widely distributed   in the tropical and subtropical regions. Their leaves, fruits and roots contain pharmacologically important metabolites such as phytosteroids, flavonoids, alkaloids and glycosides. Different Tribulus speciesare widely used in traditional folk medicine from Greek civilization, to India, China, Europe, Asia, Africa and Latin America[1].

Tribulus is of therapeutic, medicinal and pharmaceutical interest as it contains; saponins,flavonoids, alkaloids, glycosides, phytosterol, lignanamides, tannins, terpenoids, amides derivatives (cinammic) and proteins.  The most important bioactive metabolites were Steroidal saponins and flavonoids which are used for muscle building, conditioning and treatment of certain ailments [1].

The aqueous fresh extract of the whole plant contains inorganic nitrates, mostly potassium nitrate, that has diuretic effects.  The Tribulus species extract is also used for urinary dysfunction, asthma, ophthalmic and different diseases [2, 3] and has been shown to have antihypertensive properties and protection against oxidative stress and also has vasodilating properties [4, 5].  It may exhibit antitumor, cytotoxic, antifungal and anthelmintic properties [6].

In traditional medicine T. terrestris is believed to have aphrodisiac properties for treatment of male’s infertility [7]. Saponins formulation from T. terrestris were applied for veterinary production in Bulgaria, as it effectively stimulated their spermatogenesis, libido sexuality [8,9]. T. terrestris extracts increased sexual function in rats, which attributed to increase in testosterone, dihydrotestosterone, and Dehydroepiandrosterone.  Tribestan is a Bulgarian formulation widely used for treatment of infertility and libido disorders in men and women [10,11]. The furastanol saponins (protodioscin, protogracilin) from T .terrestris increased male spermatogenesis via Luteinizing Hormone which stimulated secretion of Testosterone that significantly improved sperms quality and quantity [12, 13].

 T.pentandrus   extracts were used in different pharmacological, and clinical trials including anticancer [14], ant cholinergic, antifilarial [15] anti-malarial [16] and as a CNS depressant and stimulant [17], hypoglycemic effect [18] immunologic effect, smooth muscle relaxant and stimulant activity. [19, 20].

The present study describes the chemical profiles of ethanolic extracts of roots, aerial parts and fruits of three Sudanese Tribulus species:Tribulus, T. terrestris, T.longipetalous and T.pentandru growing wild in Khartoum State, Central Sudan.

 

Materials and Methods:

Plant materials:

The roots, aerial parts (leaves and stem) and fruits from three Tribulus species;   T. terrestris, T. longipetalus and T.pentandrus were collected from the Faculty of Agriculture, University of Khartoum, Shambat, Khartoum North.

Preparation of extracts:

From each plant a sample of 20g were mixed with 120 mL of 80% ethanol in 500 mL conical flask. The conical flask was sealed tightly with aluminum foil for 10 days at room temperature. The filtrate was transferred to Petri dishes; the ethanolic crude extract was collected after evaporation.

 

 

Gas Chromatography /Mass Spectrophotometry   :

The qualitative and quantitative analysis of the sample was carried out by using GM/MS apparatus   (Model; GC/MS-QP2010-Ultra, Japan) with capillary column (Rtx-5ms-30m×0.25 mm×0.25µm). The sample was injected by using split mode, Helium as the carrier gas passed with flow rate 1.61 mL/min, the temperature program was started from 60 ͦ C with rate 10c/min to 300 ͦ C as final temperature degree with 3 min hold time, the injection port temperature was 300 ͦ C, the ion source temperature was 200 300 ͦ C  and the interface temperature was 250 ͦ C. The sample was analyzed by using scan mode in the range of m/z 40-500 charges to ratio and the total run time was 26 min.

The samples contents were analyzed and identified by comparing their retention times and mass fragmentation patterns with already deposited data in the library, of the National Institute of Standards and Technology (NIST).

 

 

 

 

 

Results and Discussion:

The chemical composition of ethanolic extract of Tribulus terrestris

GC/MS chromatogram of ethanolic extract of roots of T.terrestris (Fig.1) clearly showed 20 peaks indicating the presences of 20 compounds. The  major compounds were listed  in Table.1; sucrose (28.72%), 1-(+)–ascorbic acid,  2,6-dihexadecanoate (13.34%); alpha-d-manno-furanoside, methyl (13.18%) 9,12,15–octa decatrienoic acid (Z,Z,Z) – (8.54%); diosgenin (8.32%) and cyclopentane1-acetyl-1,2 –epoxy  (4.99%).

GC/MS analysis of aerial parts extracts of T.terrestris identified  27 compounds (Fig.2).  The main compounds were; 4-O-methylmannose (49.60%),  gamolenic acid (10.73%), 1-(+)–ascorbic acid 2,6-dihexadecanoate (10.64%),

Sucrose (6.33%) and  phytol acetate (3.33%) (Table.1).

GC/MS chromatogram of ethanolic extract of fruits of T.terrestris (Fig.3) showed the presence of 29 compounds.  The major compounds were: 4-O-methylmannose (43.66%), 1-(+) – ascorbic acid 2,6-dihexadecanoate (10.35%),  linoleic acid ethyl ester (5.94%), 5-hydroxy methyl furfural (5.37%), gamolenic acid (5.15%) and 4-o-methylmannose (4.06%) (Table.1).

Fig.1   GC/MS chromatogram                                    Fig.2     GC/MS chromatogram of

Fig.1

  1. terrestris roots T. terrestris aerial parts                                              GC/MS chromatogram of  T. terretris fruits

 

The chemical compositions of ethanolic extract of Tribulus longipetalous

GC/MS chromatogram of ethanolic extract of roots T.longipetalous (Fig.4) 23 compounds was identified.  The major compounds were (Table.1); 4-O-methylmannose (73.11%), 1-(+)–ascorbic acid 2,6-dihexadecanoate (12.75%),  sucrose (11.92%),  9,12,15 –Octadecatrienoic acid (Z,Z,Z) (5.56%), 9,12–octadecadienoic acid (Z,Z)-, methyl ester (5.30%) and  9 –octadecenoic acid methyl ester (e) (4.99%).

 

 

Twenty compounds were detected in the aerial parts of T. longipetalous (Fig.5) (Table.1).

The major compounds were; 4-o-methylmannose (43.24%), 1-(+)–ascorbic acid 2,6-dihexadecanoate (14.72%), oleic acid (10.08%),  9,12,15 octadecatrienoic acid (Z,Z,Z) – (8.76%); phytol acetate (4.69%) and octadecenoic acid (4.73%).

Analysis of T. longipetalous fruits extract detected 29 compounds (Fig.6) (Table.1). The compounds were;   4-o-methylmannose (35.75%); octadec-9-enoicacid (23.35%), palmitoleic acid (1.86%), stigmastane (5.18%); squalene (4.25%) and octadecenoic acid (4.19%).

Fig.4 G

C/MS chromatogram of                                   Fig.5 GC/MS chromatogram of

  1. longipetalous roots T.longipetalous aerial  parts

Fig.6  GC/MS chromatogram of T.longipetalous fruits

The chemical contents of ethanolic extract of Tribulus pentandrus

Thirty one compounds were identified from extracts of T.pentandrus roots (Fig.7, Table.1). The compounds were;  4-o-methylmannose (47.86%), sucrose (14.55%), 1-(+)-ascorbic acid 2,6-dihexadecanoate (8.87%), gamolenic acid (4.23%), 5-alpha-hydroxy-6-beta-methyltigogenin (3%) and diosgenin (1.46%).

Thirty one compounds were detected in aerial parts of  T.pentandrus (Fig.8, Table.1). The compounds were;  4-o-methyl mannose (44.75%); gamolenic acid (15.215%), 1-(+)–ascorbic acid 2,6-dihexadecanoate (10.14%), sucrose (6.16%) and diosgenin (2.08%).

Forty two compounds were identified in fruits of T.pentandrus (Fig.9, Table.1). The compounds were; 4-O-methylmannose (55.05%), 1-(+)–ascorbic acid 2,6-dihexadecanoate (10.97%); Linoleic acid ethyl ester (3.96%); Octadecenoic acid (3.30%); Sucrose (3.27%) and Diosgenin (1.55%) as the major phytochemical constituents.

Chemical compounds T. terrestris T.longipetalous T.pentandrus
Roots Aerial Parts Fruits Roots Aerial Parts Fruits Roots Aerial Parts Fruits
Sucrose 28.72 6.33 11.92 14.55 6.16 3.27
1-(+)–Ascorbic acid.2,6-dihexadecanoate 13.34 10.64 10.35 12.75 14.72 8.87 10.14 10.97
alpha-d-manno-furanoside,methyl 13.18
Linoleic acid 8.54 5.56 8.76
Diosgenin 8.32 1.46 2.08 1.55
cyclopentane,1-acetyl-1,2 –epoxy 4.99
4-O-Methyl mannose 49.60 43.66 73.11 43.24 35.75 47.86 44.75 55.05
Gamolenic acid 10.73 5.15 4.23 15.21
Phytol acetate 3.33 4.69
Linoleic acid ethyl ester 5.94 3.96
5-hydroxy methyl furfural 5.37
9,12–octadecadienoic

Acid (z,z)-,methyl ester

5.30  

9 – Octadecenoic acid methyl ester 4.99
Oleic acid 10.08
Stearic acid 4.37 4.19 3.30
Octadec – 9 –  enoic acid 23.35
Palmitoleic acid 1.86
Stigmastane 5.18
Squalene 4.25
5.alpha.-hydroxy-6-beta-methyl tigogenin 3.26

Fig.7 GC/MS chromatogram of                           Fig.8 GC/MS chromatogram of

Tribulus pentandrus roots                                    T.pentandrus     aerial  parts

Fig.9 GC/MS chromatogram of T.pentandrus fruit

Table.1 Chemical compositions of three Tribulus species: T. terrestris, T.longipetalous and  T.pentandrus

 

Chemical classes identified in studied samples:

All of the studied samples (aerial parts, fruits and roots) of T. terrestris, T.pentandrus and T. longipetalus contain sugar (range between 62.4% in T.pentandrus roots and 28.7% in T.terrestris roots) (Table.2). All samples (except T. terrestris roots) contain terpenes (range between 6.9% in T. longipetalus aerial parts and 0.3% in T.pentandrus roots). Tribulus pentandrus roots and T. terrestris aerial parts contain   steroids 4.2%, 3.3% respectively (Table.2).

Alkanes were identified in T. longipetalus fruits (12%) and in T.terrestris roots (4%).

Alcohols were detected in only T.terrestris (4%).

Steroidal sapogenins were identified in roots of T.terrestris, T.longipetalous and T.pentandrus  (9.5%,   5%, and  4.7%, respectively). Also alkaloids were found in roots of T.pentandrus  (1%)  T. terrestris (0.6%).

 

 

Fatty acids were detected in all species (range between 42.6% in T. longipetalus fruits and 20.8% in T.pentandrus roots). Nine fatty acid were identified T.terrestris aerial parts (29%); Gamolenic acid (10.8%) and octadecenoic acid (1.8%). Seven fatty acid components were identified. T.  terrestris fruits (32%); Linoleic acid ethyl ester (6%)  and  Gamolenic acid (5.2%). Five fatty acid components were identified in T. terrestris roots (34.4%); Linoleic acid (8.5%) and  9- octadecenoic acid (E) – (4.7%).

Five fatty acids were identified in T.pentandrus aerial parts (32.8%);  Gamolenic acid (5.2%)  and  Linoleic acid (2%). Ten fatty acid were identified in fruits of  T.pentandrus (28.5%); Linoleic acid ethyl ester (3.96%) and  Gamolenic acid (3.6%). Six fatty acids were identified in roots of  T.pentandrus (20.7%); Gamolenic acid (4.2%) and  Linoleic acid (3%).

 

 

Fatty acid T. terrestris T.pentandrus T.longipetalous
A P F R A P F R A P F R
Pentadecanoic acid 1.69 0.26
9,12 – octadecadienoic acid (z,z) – , methyl ester 0.16 0.70

 

3.31 0.99 5.30
9-octadecenoic acid (E) – 0.14 4.73 0.62 4.99
9,12,15 –octadecatrienoic acid ( z,z,z) – 0.42 8.54 2.04 0.27 8.76 5.56
Octadecanoic acid 1.80 2.90 2.33 o.88 3.30 0.83 4.73 4.91 3.00
Dodecanoic acid 0.19 0.19 0.25
Tetradecanoic acid 0.95 1.21 1.94
Hexadecanoic acid  methyl ester 0.52 1.42 0.52 0.99 0.39 0.53 0.52 0.62
Linoleic acid ethyl ester 1.60 5.94 3.96 2.96 3.87
Gamolenic acid 10.73 5.15 15.21 3.63 4.23
Oleic acid 1.32 1.32 1.83 10.08
Butyl 9,12,15 – octadeca trienoate 1.03
6 – Octadecenoic acid , (z) – 1.86
Octadeca-9-enoic acid 23.35

 

 

 

 

 

 

In aerial parts of T. longipetalus, five fatty acids were detected (42.3%); Oleic acid (10.1%) and Linoleic acid (8.8%).  In its fruits four fatty acids were detected (42.6%); Octadeca – 9 – Enoic acid (23.2%) and Octadecenoic acid (4.2%). In its roots seven fatty acid were found (36%); Linoleic acid (5.6%) and 9,12– Octadecadienoic Acid (z,z)-, methyl ester (5.3%)

Chemical class T. terrestris T.pentandrus T.longipetalous
A P F R A P F R A P F R
Sugars 55.93 47.72 28.72 50.90 58.32 62.41 43.24 35.75 49.03
Fatty acid 28.94 31.8 34.83 32.83 28.48 20.67 42.33 42.56 35.99
Terpenes 5.60 3.91 6.51 2.16 0.31 4.85 4.96 4.56
Steroids 30.30 2.75 2.54 4.23 1.07 1.77
Alkanes 0.91 4.99 1.04 1.51 2.12 12.03
Alcohols 1.59 0.61 4.07 0.75 0.24 0.91 1.14 0.84
Steroidal sapogenins 9.45 3.67 2.30 4.72 5.04
Alkaloids 0.57 0.41 0.40 0.95 0.34

Table.2   Classes of chemical compounds detected in three Sudanese Tribulus species

Table.3:  Fatty acids in T. Terrestris,     T.pentandrus and T. longipetalous

The Phytochemical compositions   of T. terrestris were extensively studied including, saponins, flavonoids, alkaloids, lignanamides and cinammic acid amides [17]. Tribulus pentandrus is rich in many potential biological substances including, steroidal saponins, flavonoids, alkaloids, amides, lignans, cinammic acid, nitrate and phytosterol [6,20,21,22,23].

 

 

 

 From Tribulus longipetalus many scientists identified;   steroidal saponins, lignin-amides, alkaloids,    spirostane, furostane and cholestane nuclei and flavonoids – flavonol glycosides [20, 21,24,25].

The steroidal sapogenin was identified from the root of the Tribulus species (Tt,Tl,Tp; 9.5,5.1 and 4.7%) in the present research. Diosgenin- steroidal sapogenin  has  wide  range  of  pharmacological  and medicinal properties  and were used for  production of steroidal drugs for treatment of hyperlipidemia, inflammation, neurological diseases  and as anticancers [20,21,26,27,28]. In this study, Tribulusterrestris  roots had higher percentage of diosgenin (8.32%), however, all samples of T.pentandrus contain diosgenin (1.46 to 2.08%).  In China saponins were reported to have cytotoxic activity against in vitro culture [29].  Another study from Korea revealed t T. terretris aqueous extract can induce apoptosis in human liver cancer cell [30].  A protective effect of Tribulus terretris   alcoholic extract was reported before [31,32,33,34]

 

Many authors noticed reduction in the serum glucose level to normal, and decrease in creatinine level in liver and blood (Amin et al 2006; El-Tantawy and Hassanin, 2007; Bankdaran et al 2008

This study identified Linoleic acid in roots of T.terrestris  (8.54%) and  roots and aerial parts of T.longipetalous   (5.56-8.76%), respectively.

Linoleic acid (gamma-linoleic acid – Gamolenic acid), is an essential source of   omega -6 and 3 polyunsaturated fatty acid.  α-Linoleic acid has  neuroprotective, anti-inflammatory, antihypertension, anticholestrol   and antidepressant properties   Various Gamolenic acid formulations were indicated for treatment of rheumatoid arthritis, atopic eczema , acute respiratory distress syndrome, asthma, cardiovascular disease, cancer and insomnia [34,35,36].

Its deficiency in diet may cause neurological and cardiological disorders [34]. Linoleic acid is a popular for preventing and treating diseases of the heart and blood vessels. It is used to prevent heart attacks, high blood pressure and cholesterol [35,36].

 Conclusion:

This study identified pharmaceutically very important bioactive substances from    the three Tribulus species. These results provide a baseline for preparation of monograph for evaluation and identification.  This study recommended the three species of Tribulus as an important source of various vital pharmaceutical phytocompone

 

 

References

[1] De Combarieu EN,  Fuzzati  M,  Lovati  A and  Mercalli E. Furostanol saponins from Tribulus terrestris. Fitoterapia., 2003; 74: 583-591

[2] Sarwat M, Das S and Srivastava PS. Analysis of genetic diversity through AFLP, SAMPL, ISSR and RAPD markers in Tribulus terrestris, a medicinal herb. Plant Cell Rep. 2008; 27: 519-528.

[3] Qureshi R,  Bhatti GR,  and  Memon RA. Ethnomedicinal uses of herbs from northern part of Nara desert, Pakistan. Pak. J. Bot. 2010; 42(2): 839-851.

[4] Phillips  SJ, Anderson RP,  and  Schapire RE. Maximum entropy modeling of species geographic distributions. Ecol. Model. 2006; 190: 231-259.

[5] Sharifi  AM,  Darabi R. and  Akbarloo N.  Study of antihypertensive mechanism of Tribulus terrestris in 2K1C hypertensive rats: role of tissue ACE activity. Life Sci. 2003; 73(23): 2963-71.

[6] Deepak M, Dipankar G, Prasanth D,  Asha MK,  Amit A. and Venkatraman BV.  Tribulosin and β-sitosterol-Dglucoside, the antihelminthic principles of Tribulus terrestris. Phytomedicine. 2002; 9: 753-756.

[7] Gauthaman K,  Adaikan PG. and Prasad RNV. Aphrodisiac properties of Tribulus terrestris  extract (protodioscin) in normal and castrated rats. Life Sci. 2002; 71: 1385-1396.

[8] Tomova M, Panova D, Zarkova S.  and  Dikova V. Bulg. Patent. 1966; (11): 11450- 61.

[9] Tomova M. and R  Gyulemetova.  Bulg. Patent. 1978; (11) 26221, 2(51) A 1K 35.

[10] Protich M,  Tsvetkov D,   Nalbanski B, Stanislavov R. and  Katsarova M. 1981. Clinical trial of Tribestan on infertile males. (Pharmachim, Bulgaria – Scientific-technical report). 1981.

[11] Viktorov I,  Kaloyanov D, Lilov AI,  Zlatanova L.  and  Kasabov V.  Clinical investigation on Tribestan in males with disorders in the sexual function. 1982. Med. Biol. Inform. (Pharmachim, Bulgaria – Company documentation).

[12] Tomova M,   Gjulemetova R, Zarkova S, Peeva S,   Pangarova T. and  Simova M. Steroidal sponins from Tribulus terrestris L. with a stimulating action on the sexual functions. In First International Conference on Chemical, Biotechnological and Biologically Active Natural Products, Proceedings Varna Bulgaria, September 21—26. 1983;  298-302.

[13] Brown AG,  Vukovich MD, Martini ER, Kohut ML, Franke WD, Jackson DA.  and King DS. Endocrine and lipid responses to chronic androstenediolherbal supplementation in 30 to 58 year old men. J. Am. Coll. Nutr. 2002; 20: 520-528.

[14] Sun  B, Qu  W. and Bai Z. The inhibitory effect of saponins from Tribulus terrestris on Bcap- 37 breast cancer line in vitro. Zhong Yao Cai. 2003; 26: 104-106.

[15] Comley JCW, Titanji VPK, Ayafor JF.  and Singh VK.  In vitro antifilarial activity of some medicinal plants. Acta Leide-nsia. 1990; 59(12): 361-363

[16] Misra P, Pal NL, Guru PY, Katiyar JC.  and Tandon JS. Antimalarial activity of traditional plants against erythrocytic stages of Plasmodium berghei. Int. J. Pharmacog. 1991; 29(1): 19-23.

 

[17] Li M, Qu W, Wang Y, Wan  H. and Tian C. Hypoglycemic activity of saponin from Tribulus terrestris. Zhong Yao Cai. 2002; 25: 420–422.

[18] Arcasoy  HB, Erenmemisoglu A, Tekol Y, Kurucu S. and Kartal M.  Effect of Tribulus terrestris L. saponin mixture on smooth muscle preparations: a preliminary study. Boll. Chim. Farm. 1998; 137: 473-475.

[19] Ross IA. Medicinal plants of the world: chemical constituents, traditional and modern medicinal uses. Humana press Inc. Totowa. 1996; 2: 411- 427.

[20] Ruxandra S,  Tefanescu, RS,  Tero-Vescan A,   Negroiu A,    Aurica E. and Vari CE. A Comprehensive Review of the Phytochemical, Pharmacological, and Toxicological Properties of Tribulus terrestris L. Biomolecules. 2020; 10, 752-65.

doi:10.3390/biom10050752      www.mdpi.com/journal/biomolecules

[21] Semerdjieva IB. and  Zheljazkov VD. Chemical Constituents, Biological Properties, and Uses of Tribulus terrestris: A Review. Nat. Prod. Commun. 2019; 14, 1–26.

DOI: 10.1177/1934578X19868394  journals. sagepub. com/ home/ npx

 

[22] Saleh NA, Ahmed AA, Abdalla MF. Flavonoid glycosides of Tribulus pentandrus and T.terrestris.  Phytochemistry. 1982; 21(8):1995-2000.

 

[23] Wu KL, Kang LP, Xiong CQ, et al. Study on chemical components of steroidal saponins from Tribulus terrestris L. J Tianjin Univ Tradit Chin Med. 2012; 31, 225-228.

 

[24] Wang Y, Ohtani K, Kasai R, Yamasaki K. Steroidal saponins from fruits of Tribulus terrestrisPhytochemistry. 1997; 45(4):811-817.

 

[25]  Wang, Z.F.;Wang, B.B.; Zhao, Y.;Wang, F.X.; Sun, Y.; Guo, R.J.; Song, X.B.; Xin, H.L.; Sun, X.G. Furostanol and Spirostanol Saponins from Tribulus terrestris. Molecules. 2016; 21, 429.

 

[26] Cai LE, Jing FY, Zhang JG, et al. Studies on the chemical components of Tribulus terrestris. Yao Xue Xue Bao. 199; 759-761.

 

[27] Cai L, Wu Y, Zhang J, et al. Steroidal saponins from Tribulus terrestris. Planta Med. 2001; 67(2):196-198.

 

[28]  Hu K, Dong A, Yao X, Kobayashi H, Iwasaki S. Antineoplastic agents; I. Three spirostanol glycosides from rhizomes of Dioscorea collettii var. hypoglauca. Planta Med. 1996; 62(06):573-575

 

[29] Tang  YN, Pang YX,  He  XC,  Zhang YZ,  Zhang JY,  Zhao  ZZ,  Yi  T,  Chen  HB. UPLC-QTOF-MS. identification of metabolites in rat biosamples after oral administration of Dioscorea saponins: A comparative study. J. Ethnopharmacol. 2015; 165, 127–140.

 

[30] Kim  HJ,  Kim  JC,  Min JS,  Kim MJ,  Kim  JA, Kor  MH,  Yoo HS.and   Ahn  JK. Aqueous extract of Tribulus terrestris Linn induces cell growth arrest and apoptosis by down-regulating NF-_B signaling in liver cancer cells. J. Ethnopharmacol. 2011; 136 : 197–203.

 

[31]   Amin AMR, Lotfy M, Shafiullah M, Adeghate E. The protective effect of Tribulus terrestris in diabetes. Ann N Y Acad Sci. 2006;1084(1):391-401.

 

[32] El-Tantawy WH, Hassanin LA. Hypoglycemic and hypolipidemic effects of alcoholic extract of Tribulus alatus in

streptozotocin-induced diabetic rats: A comparative study with T. terrestris (Caltrop). I J Exper Biol. 2007;45:785-790.

[33]  Bonakdaran A, Hosseini F, Khalighi H, Sigaroudi F, Ahvazi M. Investigation of the hypoglycemic effect of Tribulus terrestris extract on diabetic rats. J Med Plants. 2008;7:85-92.

 

[34] Cunnane SC. and   Anderson MJ. Pure linoleate deficiency in the rat: influence on growth, accumulation of n-6 polyunsaturates, and [1-14C]linoleate oxidation. Journal of Lipid Research. 1997; 38(4), 805—12.  https://doi.org/10.1016/S0022-2275(20)37247-3.

 

 

[35] Blondeau N,  Lipsky  RH,  Bourourou M,   Duncan MW,  Gorelick PB. And  Marini AM. “Alpha-Linoleic acid: An Omega-3 Fatty Acid with Neuroprotective Properties—Ready for Use in the Stroke Clinic?”, BioMed Research International. 2015;   https://doi.org/10.1155/2015/519830

[36] Zhao WR, Shi WT, Zhang J, Zhang KY, Qing Y, Tang JY, Chen XL, Zhou ZY. Tribulus terrestris L. Extract Protects against Lipopolysaccharide-Induced Inflammation in RAW 264.7 Macrophage and Zebrafish via Inhibition of Akt/MAPKs and NF-κB/iNOS-NO Signaling Pathways. Evid Based Complement Alternat Med. 2021; 6628561.  doi: 10.1155/2021/6628561. PMID: 33628304; PMCID: PMC7895590.

4 Tribulus

 10 total views