# Preclinical Evaluation of the Anticoagulant Effect of an Aqueous Extract of Snow Mountain Garlic
## 4 Discussion  
Plants and plant foods have been shown to have promising results in aiding in the treatment of thrombosis [18]. According to the WHO, 80% of the population uses traditional medicine as a primary care option for various diseases [11]. Medicinal plants represent the ancestral wisdom of people around the world. They contain many metabolites with therapeutic effects. S. mountain garlic is commonly used for pain therapy in villages near the Himalayas [29]. This work validated that the LAE of S. mountain garlic has an anticoagulant effect _in vivo_. An increased bleeding time was observed with 500 and 1000 mg/kg, which showed significant effects at 1, 2, and 4 weeks, with the strongest effects observed at 2 and 4 weeks (\(p<0.05\)). The ASA-treated group demonstrated significant effects at 30 min and 2 weeks. However, from week 8, the pairwise effects began to diminish across all groups; however, the differences among groups remained statistically significant at both weeks 8 and 12. Similarly, a universal anticoagulant effect (AUC) was shown during the 90 days with the dose of the LAE at 500 mg/kg and 100 mg/kg of ASA. In contrast to ASA, which resulted in duodenal ulcers, acanthosis, and lymphoplasmacytic inflammatory infiltrate, doses below 500 mg showed no histopathological alterations. However, at doses of 1000 mg/kg and 2000 mg/kg, histopathological changes similar to those induced by ASA were observed. The ED\({}_{50}\)_in vivo_ was 366 \(\pm\) 2.43 mg/kg. Most work that has been conducted to evaluate the clotting time or bleeding _in vivo_ of garlic _Allium sativum_ L. has used ethanol extracts, garlic oils, and garlic powder, among others, with doses of 10 mg/kg/d, 50 mg/kg/d, and 1000 mg/mL, respectively, in short periods of 2 to 4 weeks; these show a wide variety of doses and extracts whose results are not conclusive regarding their use in humans [20]. On the other hand, in our work, doses of 250 mg/kg/d and 500 mg/kg/d had no histopathological findings in tissues; results like those found by [PERSON] et al., 2015, with the administration of 200 mg/kg of _Allium sativum_ L, showed that it has hepatoprotective activity against the toxic action of paracetamol [34]. Nine organosulfate compounds were identified using GC-QToF-MS. Diallyl disulfide (DADS) and diallyl trisulfide (DATS) are the most-studied compounds; evidence has been found to show that these have an inhibitive effect on platelet aggregation [35; 36]. On the other hand, allyl methyl trisulfide is documented to have an antiplatelet effect, but this is less documented than it is for DADS and DATS [37]. Diallyl sulfide is a bioactive compound of common garlic, but its anticoagulant activity is not as well documented as it is in the cases of DADS and DATS. On the other hand, a study by [PERSON] 2022 identified the sulfurous compounds of the garlic S. mountain vs. the common garlic (_Allium sativum_ L.); they identified new organosulfate compounds that were not found  
Figure 4: GC-QToF-MS of the organosulfate compounds of S. mountain garlic. The compounds of each peak are described in Table 2.  
in _Allium sativum_ L.: methanethiol, dimethyl sulfide, propyl mercaptan, 1-allyl-2-isopropyldisulfide, pentadecanoic acid, and Cholesta-4-6-dien-3-ol,(3-beta). In our work, new organosulfur compounds were identified that had not been described for Snow mountain garlic: Thiophene 3,4-dimethyl, Methyl (E)-1-propenyl disulfide, (Z)-1-allyl-2-(prop-1-en-1-yl) disulfan, and Allyl prop-1-enyl disulfide [25]. Novel oral anticoagulants (NOACs) have been in clinical use for several years; however, significant knowledge gaps remain regarding their optimal utilization [38]. This underscores the importance of ongoing research and development of new anticoagulants to enhance patient outcomes and minimize adverse effects. Medicinal plants, such as S. mountain garlic, present an excellent option in this context.