Research in Plant Metabolites

Research in Plant Metabolites

Identification of Secondary Metabolites in Malva sylvestris Extract Using LC/MS/MS and Its Impact on Milk Quality of Najdi Dairy Goats

Document Type : Original Article

Authors
Hossein Noori Noroozi
Tahereh mohammadabadi
Morteza Chaji
Agricultural Sciences and Natural Resources University of Khuzestan, Iran
10.22034/jrpsm.2026.553034.1097
Abstract
Background and Objective
In recent years, the use of natural plant additives in animal nutrition has attracted attention as a safe and sustainable alternative to chemical additives. This approach is particularly important in lactating ruminants. Because milk quality depends not only on the dietary ration of the animal but also on its antioxidant status and microbial load. The mallow plant (Malva sylvestris) is native to various regions of Iran and contains various bioactive components such as phenolic compounds, flavonoids, anthocyanins, and alkaloids. These compounds have antioxidant, anti-inflammatory and antimicrobial properties and can play an effective role in improving the animal health and quality of animal products.
There is limited information about the nutritional application of mallow extract on milk quality. Therefore, the objective of this study was to identify the secondary metabolites of the ethanolic extract of the mallow plant using the LC/MS/MS method and to investigate the effect of adding this extract to the diet of lactating Najdi goats on milk antioxidant capacity and microbial load.
Materials and Methods
The mallow plant was collected at the flowering stage. After drying the samples in the shade, extraction was performed using the maceration method with 70% ethanol at a ratio of 1:10 (weight/volume) for 24 hours at room temperature. Following filtration, the solvent was removed using a rotary evaporator, and the final extract was stored at 4 °C until further analysis. Identification of the bioactive compounds of the extract was carried out using an LC–MS/MS system (Waters, USA). For separation of the compounds, a C18 column and a mobile phase consisting of 1% formic acid in water and acetonitrile were used. The obtained data were analyzed using MassLynx software.
In the animal section of the experiment, twelve lactating Najdi goats with an average body weight of 26 ± 4 kg were divided into three treatments in a completely randomized design: control group: basal diet (without extract addition), treatment one: basal diet + extract at 0.2% of daily dry matter intake, and treatment two: basal diet + extract at 0.4% of daily dry matter intake. Milk antioxidant capacity was measured using the DPPH assay, and milk microbial load was measured using the culture method on Plate Count Agar medium and colony counting.
Results
The results obtained from LC/MS/MS analysis showed that the ethanolic extract of the mallow plant contained 11 phenolic compounds, 4 anthocyanins, and 7 alkaloids. The identified phenolic compounds included gallic acid, benzoic acid, syringic acid, 3,4‑dimethoxycinnamic acid, p-coumaric acid, myristicin, caffeic acid, luteolin, chlorogenic acid, and apigenin. The highest concentrations were for p-coumaric acid and 3,4‑dimethoxycinnamic acid. The identified anthocyanins included malvidin‑3‑O‑glucoside, cyanidin‑3‑glucoside, delphinidin‑3‑O‑glucoside, and cyanidin‑3‑rutinoside. In addition, alkaloids such as tetrandrine and cocsuline were identified which have been reported in previous studies to exhibit antimicrobial and anti‑inflammatory properties. Supplementation of mallow extract in the diet of lactating goats caused a significant increase in milk antioxidant capacity compared with the control group, such that the highest value was observed in the treatment receiving 0.4% extract. Moreover, the milk microbial load in the treatments containing mallow extract was significantly reduced.
Conclusion
Based on the results of this study, the ethanolic extract of the mallow plant is rich in phenolic, anthocyanin, and alkaloid compounds with antioxidant and antimicrobial properties. Adding this extract to the diet of lactating Najdi goats improved milk antioxidant capacity and reduced milk microbial load. Therefore, mallow extract may be used as an effective natural additive in the nutrition of lactating goats in order to improve the nutritional and hygienic quality of milk.
Keywords
Mallow (Malva sylvestris)
plant extract
LC/MS/MS
milk antioxidant capacity
microbial load
Al-Johani, N. S., Aytah, A. A., Boutraa, T., & Arabia, S. (2012). Allelopathic impact of two weeds, Chenopodium murale and Malva parviflora on growth and photosynthesis of barley (Hordeum vulgare L.). Pakistan Journal of Botany,44(6),1865–1872.
Banakar, P. S., Kumar, S., Vinay, V., Dixit, S., Tyagi, N., & Tyagi, A. K. (2021). Supplementation of Aloe vera extract in lactating goats’ diet: effects on rumen fermentation efficiency, nutrient utilization, lactation performance, and antioxidant status. Tropical animal health and production, 53, 1-10.
Batiha, G. E.-S., Tene, S. T., Teibo, J. O., Shaheen, H. M., Oluwatoba, O. S., Teibo, T. K. A., . . . Papadakis, M. (2023). The phytochemical profiling, pharmacological activities, and safety of malva sylvestris: a review. Naunyn-Schmiedeberg's archives of pharmacology, 396(3), 421-440.
Burgos-Briones, G. A., Verano-Naranjo, L., Cejudo-Bastante, C., Dueñas-Rivadeneira, A. A., Mantell-Serrano, C., & Casas-Cardoso, L. (2023). Extraction of Bioactive Compounds from Prestonia mollis Leaves and Their Impregnation into Polylactic Acid Using High-Pressure Technologies: Potential for Biomedical Application. Antioxidants, 12(10), 1864.
Çam, M., Hışıl, Y., & Durmaz, G. (2009). Classification of eight pomegranate juices based on antioxidant capacity measured by four methods. Food chemistry, 112(3), 721-726.
Demirtaş, A. (2021). Evaluation of the stimulatory and inhibitory effects of Malva sylvestris leaf extract on some beneficial and pathogenic bacteria from the colon. Journal of Istanbul Veterinary Sciences, 5(1), 13-18.
Fathi, M., Ghane, M., & Pishkar, L. (2022). Phytochemical composition, antibacterial, and antibiofilm activity of Malva sylvestris against human pathogenic bacteria. Jundishapur Journal of Natural Pharmaceutical Products, 17(1).
Gasparetto, J. C., Martins, C. A. F., Hayashi, S. S., Otuky, M. F., & Pontarolo, R. (2012). Ethnobotanical and scientific aspects of Malva sylvestris L.: a millennial herbal medicine. Journal of Pharmacy and Pharmacology, 64(2), 172-189.
Hervás, G., Frutos, P., Giráldez, F. J., Mantecón, Á. R., & Del Pino, M. a. C. Á. (2003). Effect of different doses of quebracho tannins extract on rumen fermentation in ewes. Animal Feed Science and Technology, 109(1-4), 65-78.
Jimoh, O. A., Daramola, O. T., Okin-Aminu, H. O., & Ojo, O. A. (2022). Performance, hemato-biochemical indices and oxidative stress markers of broiler chicken fed phytogenic during heat stress condition. Journal of Animal Science and Technology, 64(5), 970.
Kakabouki, I., Karydogianni, S., Roussis, I., & Bilalis, D. (2020). Effect of organic and inorganic fertilization on weed flora and seed yield in black mustard [Brassica nigra (L.) Koch] crops. Ciencia e investigación agraria: revista latinoamericana de ciencias de la agricultura, 47(2), 2.
Khalaji, S., Zaghari, M., & Nezafati, S. (2011). The effects of mannan-oligosaccharides on cecal microbial populations, blood parameters, immune response and performance of broiler chicks under controlled condition. African Journal of Biochemistry Research, 5(5), 160-164.
Mohammadabadi, T., Hoseini, S. (2022). Effect of Malva sylverstris plant on milk quality and production, liver enzymes and nutrients digestibility of Khuzestani Buffalo. Journal of Ruminant Research, 9(4), 109-120.
 Mohan, E., Suriya, S., Shanmugam, S., Muthupandi, C., & Rajendran, K. (2021). Preliminary phytochemical appraisal of selected medicinal plants.
National Research Council (US). Committee on Nutrient Requirements of Small Ruminants. (2007). Nutrient requirements of small ruminants: Sheep, goats, cervids, and new world camelids. The National Academies Press.
Salahvarzi, M., Nasr Esfahani, M., Shirzadi, N., Burritt, D. J., & Tran, L. S. P. (2021). Genotype‐and tissue‐specific physiological and biochemical changes of two chickpea (Cicer arietinum) varieties following a rapid dehydration. Physiologia plantarum, 172(3), 1822-1834.
Shadid, K. A., Shakya, A. K., Naik, R. R., Jaradat, N., Farah, H. S., Shalan, N., . . . Oriquat, G. A. (2021). Phenolic content and antioxidant and antimicrobial activities of Malva sylvestris L., Malva oxyloba Boiss., Malva parviflora L., and Malva aegyptia L. leaves extract. Journal of Chemistry, 2021(1), 8867400.