HUM-27, known as Malviya Jankranti, is a high-protein (28.9%) Mung Bean variety developed by the Department of Genetics and Plant Breeding, Banaras Hindu University, Varanasi, with bold seeds (5 g/100 seeds), an average plant height of 44 cm and a maturity period of 62-70 days. This variety is highly resistant to Mung bean yellow mosaic virus and tolerant of high temperatures (>45°C). The Government of India released HUM-27 (Fig 1) and it is widely adopted for both kharif and spring cultivation in the Central Zone (Madhya Pradesh and Chhattisgarh).


       
Since the Vedic period, Mudga has been regarded as one of the most important pulses in Ayurveda. Bhavaprakasha Nighantu (2023) describes five types based on colour: black (Shyama), green (Harita), yellow (Peeta), white (Sweta) and red (Rakta). Mudga is considered the best among pulses and is recommended for regular consumption due to its easy digestibility (Charak Samhita,  2023). Its therapeutic properties are described in terms of Shadrasa, Virya and Vipaka. It is traditionally indicated as Yusha for conditions such as fever, ascites, cough and Vatarakta (hyperuricemia) and as Pathya Ahara after Panchakarma and in the management of obesity (Sthaulya) and diabetes (Prameha). Mung beans are a rich source of easily digestible protein, Iron, Calcium, dietary fibre, phytochemicals and flavonoids, positioning them as a valuable alternative functional food, beyond their basic nutritional profile. Supplementing with mung beans exhibits a wide range of pharmacological properties, including hepatoprotective, antimicrobial, cardioprotective, anti-inflammatory, antidiabetic, anticancer, anti-obesity and hypolipidemic activities (Shen et al., 2022; Amare et al., 2022). Various therapeutic effects of mung bean (Vigna radiata), named as Mudga in Ayurveda, have been documented in both ancient Ayurvedic texts and recent scientific studies, with variations among cultivars.
 

This Research study was approved by the Institutional Ethics Committee (IAEC/6229). Experimental work was conducted (duration: July 2024-2025) at the Department of Kriya Sharir, IMS, BHU and analytical assessment of HUM-27, germinated HUM-27 and commercial mung bean samples was performed at the Department of Dairy Science and Food Technology, BHU. All reagents were of analytical grade.
 
Procurement of mung bean
 
Seeds of mung bean variety HUM-27 were obtained from Department of Genetics and Plant Breeding, Institute of Agricultural Sciences, BHU, Varanasi, India, in April 2024. HUM-27 was selected for its high protein content and evaluated for physicochemical characteristics and hypolipidemic and anti-hyperuricemic effects in an experimental rat model. Seeds were manually cleaned, stored in airtight polyethylene containers at room temperature (27°C) and used for further analysis. A commercial mung bean variety was purchased from a local market in Varanasi for comparative analysis. The sample preparation protocol is presented in Fig 2.


 
Soaking, germination and preparation of powder
 
A 150 g sample of mung bean seeds (HUM-27) was soaked in water (1:5, w/v) at room temperature for 8 h, thoroughly washed and then kept for germination under a cotton cloth for 24 h, dried in a hot air oven (45°C for 4-5 hours), then ground into powder.
 
Physicochemical analysis of mung bean powder                                    
Organoleptic test
 
Sensory evaluation was conducted by 15 semi-trained panellists (aged 18-35 years) using a 9-point Hedonic scale. Samples were evaluated for appearance, consistency, flavour, mouthfeel and overall acceptability (Johnson, 2021).
 
Proximate analysis
 
Grounded powder characteristics and proximate analysis and content determination, including crude protein, fat, fibre, moisture, ash and CHO, were determined using the standard procedures (AOAC, 2000). The energy content (EC) was determined using Atwater conversion factors applied to the respective amounts of protein, fat and carbohydrates (Kemal et al., 2025).
 

EC = (9 x % fat) + (4 x % CHO) + (4 x % proteins) + (2 x % fibre)

 
                                                                   
 Determination of techno-functional properties
 
Determination of water absorption capacity (WAC) and Oil absorption capacity (OAC)-was determined according to Brishti et al., (2017). WAC and OAC were expressed as g water/g sample and g oil/g sample, respectively.
 
Determination of swelling power (SP)- The swelling power was determined by the method given by Pranoto et al., (2014). 
 
Emulsion activity (EA) and emulsion stability (ES)- To determine emulsifying properties, 0.5 g of Mung Bean flour was suspended in 3 mL of distilled water and 3 mL of vegetable oil. After 5 minutes of vigorous agitation, the mixture was centrifuged at 2000 rpm for 30 minutes to separate the emulsion layers.
 
Enzymatic activity
 
Amylase and protease extracts were prepared by suspending 2 g sample in 10 mL distilled water, shaking for 1 h at room temperature and centrifuging at 10,000 rpm for 10 min, while lipase was extracted from 100 g sample with 40 mL 30% (v/v) acetone for 3 h at 4°C, followed by filtration and centrifugation at 10,000 ´g for 15 min (Sattar et al., 2017). Amylase activity was assayed using soluble starch (1%) in acetate buffer (0.1 M, pH 4.5) at 37°C for 1 h, followed by DNS treatment and absorbance measurement at 550 nm. Protease activity was determined using casein (0.65%) in Tris-HCl buffer (50 mM, pH 7.4) at 37°C for 30 min, with TCA precipitation and folin-ciocalteu assay at 660 nm and expressed as μg tyrosine released/min. Lipase activity was determined by incubating the extract with olive oil, acetate buffer (50 mM, pH 5.6) and CaCl‚  at 30°C for 30 min, followed by titration of liberated fatty acids with 50 mM KOH (Sattar et al., 2017).
 
Vitamin C titration method
 
Vitamin C levels were determined using a Sawhney (1996) 2,6-dichlorophenolindophenol method, with the results expressed in milligrams of ascorbic acid/100 gm.
 
LC-MS analysis of phytochemicals
 
Flavonoids and phenolic compounds were identified and quantified following Alkhatib et al., (2022), with minor modifications. An Agilent 1200 LC coupled with an agilent 6400 triple quadrupole MS (Agilent Technologies, USA) was used in positive ion mode (M+H). Mung bean powder (0.5 g) was extracted with 10 mL methanol by soxhlet extraction for 24 h. The extract was concentrated at 40°C using a rotary evaporator, reconstituted in 1 mL methanol and centrifuged at 12,000 rpm for 6 min. The supernatant was analysed by LC-MS using a C18 reversed-phase column at 55°C. Mobile phase a comprised water/methanol (90:10, v/v) with 5 mM ammonium formate and 0.1% formic acid, while mobile phase B comprised methanol with the same additives.
 
High-Performance liquid chromatography analysis of purine
 
Purine content was determined by HPLC following Fukuuchi et al., (2013), with slight modifications. Mung bean powder (0.5 g) was hydrolysed with 70% perchloric acid at 90-95°C for 60 min to release adenine, guanine, hypoxanthine and xanthine. The hydrolysate was cooled and neutralised with KOH to approximately pH 4.0, followed by centrifugation to remove potassium perchlorate precipitates. The supernatant was filtered through a 0.45 µm syringe filter and analysed using a reversed-phase C18 column with methanol and aqueous buffer as the mobile phase and UV detection at 254 nm (Feng et al., 2023).

Mineral analysis
 
Mung bean powder (0.5 g) was wet-digested with 8 mL HNO3 and 1 mL HCl on a hot plate for 30-45 min until clear. The digest was diluted to 50 mL with distilled water and analysed by atomic absorption spectroscopy (AAS) at element-specific wavelengths (Zafar et al., 2023).
 
Fourier-transform infrared spectra (FTIR)
 
FTIR spectra of mung bean powder were recorded using a Spectrum Two FTIR spectrophotometer (PerkinElmer). The sample was mixed with KBr, pelletized and scanned over 4000-400 cm^-1 after recording the background signal. The characteristic peaks were interpreted to identify the corresponding functional groups (Thummajitsakul et al., 2023).
 
DPPH (2,2-diphenyl-1-picrylhydrazyl) assay
 
DPPH is a stable nitrogen-centred radical widely used to test the free radical scavenging ability of various samples. DPPH (100 μmol L^-1) was dissolved in 96% ethanol. A 1 mL solution of DPPH and 1 mL of the mung bean water extract were mixed. After being shaken, the mixture was left to stand at room temperature in the dark for 10 min. Finally, the decrease in absorbance of the resulting solution was measured at 517 nm after 10 min. The results are reported in μmol of Trolox equivalents (TE) per gram (Shi et al., 2016).
 
Statistical analysis
 
The proximate analysis experiment was conducted in triplicate and the results are reported as the Mean±SD. Statistically significant differences between the means were determined by an Independent T-Test at a 95% confidence level. 

HUM-27 and commercial mung beans recorded the highest overall acceptability scores, indicating greater consumer preference than germinated mung beans (Table 1).


 
Physico-chemical analysis
 
Table 2 shows that mung bean variety and processing significantly influenced proximate composition. Moisture content ranged from 9.07% to 10.63%, with the lowest value observed in HUM-27 (9.07%). Protein and carbohydrates constitute the primary nutritional components in all varieties of mung bean. The highest protein content, 28.73%, was found in mung bean HUM-27 compared to the commercially available mung beans. This difference was statistically significant (P<0.0091). In the germinated HUM-27 sample, the protein content increased by 2%, accompanied by a significant rise in moisture content.  The fat content ranged from 2.37% to 3.33%, with germination resulting in reduced fat content levels (2.48%).  The carbohydrate content ranged from 48.47% to 56.48%, with the commercial mung bean showing the highest CHO content. CHO levels decreased slightly during germination, likely due to the energy expenditure associated with seedling growth. This decrease implies that germinated mung beans likely possess a marginally lower caloric density relative to ungerminated seeds (Kavitha and Parimalavalli, 2014). The energy value ranges from 342.16 kcal to 357.07 kcal/100 gm, with a significant decrease in total energy in the germinated sample.

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Techno-functional properties of mung bean
 
The germinated sample showed an enhanced water absorption capacity ranging from 2.29 to 3.01 g. Emulsion activity ranged from 43.06 to 53.23 ml and was highest in the germinated sample. The emulsion stability was 31 ml more than HUM-27 and the commercial mung bean. Swelling power and oil absorption were found to be highest in HUM-27 and the commercial sample than the germinated sample (Table 3). 

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Enzymatic activity
 
The amylase, protease and lipase activity are shown in Fig 3. The germinated sample showed the highest amylase activity (1.3 glucose U/ml), Protease activity (1.2 U/ml and lipase activity 2.3 U/ml). The lowest amylase activity was found in commercial and the highest lipase activity was in germinated HUM-27.   

     
               
Identification of phenolic compounds in mung bean
 
In the three Mung bean samples, the identified phenolic acids were five hydroxycinnamic acids (Sinapic acid, chlorogenic acid, ferulic acid, coumaric acid, caffeic acid) and five hydroxybenzoic acids (Vanillic acid, Syringic acid, gentisic acid, Ellagic acid) (Fig 4 and Table 4). In the germinated HUM-27 sample, some phenolic compounds were increased, such as gallic acid, ferulic acid, and ellagic acid, while other compounds, like sinapic acid, chlorogenic acid, p-coumaric acid, vanillic acid and syringic acid, were reduced. The most abundant phenol in HUM-27, germinated HUM-27 and commercial mung beans was sinapic acid (259.61 ng/ml, 171.72 ng/ml and 183.62 ng/ml, respectively), followed by chlorogenic acid, vanillic acid, and syringic acid.




 
Identification of flavonoids in mung bean
 
The flavonoid compounds found in all three samples were cyanidin-3-glucoside, myricetin, quercetin, kaempferol, catechin, vitexin, isovitexin and formononetin. The most abundant compounds were catechin, cyanidin-3-glucoside and isovitexin, but in the germinated HUM-27, kaempferol, catechin, vitexin and formononetin compounds decreased. (Fig 5 and Table 5).




 
Purine content
 
The maximum purine compound found was hypoxanthine, 48.634, 52.634, 41.532 mg/100 g in all three samples. The highest total Purine content was found in HUM-27 (128.16 mg), followed by germinated HUM-27 (104.55 mg) and commercial mung beans (93 mg). After germination, purine content decreased, indicating that germination reduces purine content (Mubarak et al., 2005). The maximum purine compound found was Hypoxanthine (48.634, 52.634, 41.532) in all three samples (Fig 6 and Table 6).




 
Functional compounds
 
FTIR (Fourier-transform infrared spectroscopy) is a technique for measuring the wavelength of a sample and identifying the peak values and characteristic bands of different functional groups. FTIR spectroscopy has revealed characteristic peak values for O-H stretching (Hydroxyl group), C=O stretching (Alkyl group) and C-H stretching (Carbonyl group) in all three samples, but the intensities differed among them. Germination causes biological changes, breaking down complex starches into simple sugars; these groups are responsible for their medicinal properties. The values obtained for the Mung Bean sample are shown in Fig 7.


 
Minerals
 
Phosphorus, potassium, magnesium and calcium are the predominant minerals in all three mung bean samples, with the highest content in commercial mung bean, followed by HUM-27, germinated (HUM-27). Subsequently, after germination, calcium, manganese and magnesium increased in germinated HUM-27. Iron content (110.305 mg/kg) was highest in HUM-27 (Table 7).


 
Antioxidant activity (%DPPH Inhibition)
 
The percentage of DPPH scavenging activity found in Germinated HUM-27 was 78.32%, HUM-27 and 60.32% and in the commercial sample, 60.21%, showing (Fig 8) that the antioxidant activity is maximum in germinated HUM-27. Our findings align with a study by Kemal et al., (2025).


       
The results of the present study demonstrate significant variations in proximate composition, phytochemical profile, mineral content and antioxidant activity among HUM-27, germinated HUM-27 and a commercial sample, highlighting the substantial impact of genotype and processing (germination) on nutritional and functional quality. In proximate analysis, HUM-27 showed the highest protein content (28.73%) compared to the commercial sample, highlighting the superior nutritive value of the HUM-27 genotype. Germination marginally increased protein and fibre while reducing carbohydrate and fat content. The increase in moisture was due to water absorption and enzymatic activation, while reduced fat resulted from lipase-mediated lipid mobilisation during germination (Gan et al., 2017). Research findings reported that protein content is influenced by species, genotypes, cultivars, soaking and germination. Significant changes in seed protein content occur during germination, although the magnitude varies (Zhao et al., 2022; Sen et al., 2017; Kamalasundari et al., 2019). Moisture content increased by 39% in germinated HUM-27. The highest fat content (3.33%) was found in HUM-27 mung bean, which was reduced by germination (2.48%)
       
Techno-functional properties, including WAC, OAC, SP, EA and ES, varied among the samples. Germination significantly increased WAC, possibly due to increased dietary fibre, whereas non-germinated samples showed higher SP, likely owing to greater starch availability. EA and ES ranged from 43.06-53.23 mL and 28.00-31.40 mL, respectively, with germinated samples showing the highest values (Sattar et al., 2017). Germination improved emulsification by unfolding polypeptides, exposing hydrophobic sites and increasing protein surface area. It also enhanced enzymatic activity by mobilising seed reserves. The germinated sample showed increased amylase (1.3 glucose U/mL), protease (1.2 U/mg) and lipase (2.2 U/mL) activities, consistent with previous findings (Savitha Gujjaiah et al., 2013). Vitamin C content increased in the germinated sample by 23.14 mg compared to the non-germinated sample.
       
Polyphenolic profiling revealed that ungerminated HUM-27 possessed higher concentrations of several phenolic acids, such as Sinapic, chlorogenic, vanillic and p-coumaric acids, whereas germination selectively enhanced ferulic, gallic and ellagic acids. This shift suggests enzymatic hydrolysis of bound phenolics and de novo synthesis during germination, as reported in legumes and cereals (Zou et al., 2019 and Li et al., 2012). The commercial sample showed comparatively lower and less balanced phenolic composition, likely due to varietal differences and post-harvest processing losses. The higher levels of soluble phenolics during germination likely reflect de novo synthesis and compound transformation. Flavonoid analysis further corroborated the functional advantage of germination. Germinated HUM-27 showed pronounced increases in myricetin and quercetin, compounds known for strong free-radical scavenging and anti-inflammatory activity (Panche et al., 2016). The present study identified a 18.4% reduction in purine content in HUM-27 following germination.  As elevated dietary purine intake is associated with hyperuricemia, germinated HUM-27 may offer improved dietary suitability for susceptible populations. The observed decrease supports earlier evidence indicating that soaking and sprouting lower purine levels through leaching and enzymatic degradation processes (Kaneko et al., 2014). Mung beans are a source of plant-based food classified in the low Purine content group (50-100 mg/100 g). Mineral analysis revealed that HUM-27 and germinated HUM-27 were particularly rich in iron and calcium, whereas the commercial sample showed higher potassium and magnesium. Antioxidant activity was highest in germinated HUM-27, despite some reductions in individual phenolics. This indicates a synergistic antioxidant effect, where qualitative changes in phenolic and flavonoid composition outweigh mere quantitative abundance (Desta et al., 2024). The superior antioxidant capacity of germinated HUM-27 reinforces the functional relevance of germination as a low-cost, traditional bioprocessing technique. Therefore, mung bean sprouts, rich in antioxidant phytochemicals, may be regarded as a valuable functional food that supports health, aligning with the Ayurveda description of Mudga as light (laghu), wholesome (pathya) and conducive to health promotion (Caraka SaChitâ, Sûtrasthâna).
       
Overall, the findings establish HUM-27 as a nutritionally superior genotype and demonstrate that germination significantly enhances its functional and antioxidant potential, surpassing both the ungerminated form and the commercial sample. These results support adding germinated HUM-27 to functional foods and align with traditional dietary practices emphasising sprouted legumes for improved digestibility and vitality.

The results demonstrated that nutrient bioavailability varied with mung bean variety and germination. HUM-27 exhibited the highest protein content (28.73%), along with greater levels of phosphorus, potassium, calcium and iron than germinated HUM-27 and commercial mung bean. Germination reduced fat and total purine content while enhancing protein (~20%), vitamin C, enzymatic activity and techno-functional properties. Among purines, hypoxanthine was predominant, with total purine content decreasing from 128.16 mg/100 g in non-germinated HUM-27 to 104.55 mg/100 g after germination. Higher protein, phenolic and flavonoid contents, along with higher DPPH activity, indicate that HUM-27 is a promising nutrient-and antioxidant-rich mung bean variety, making it a better source of nutrients and antioxidants than other varieties available in Uttar Pradesh.

The authors gratefully acknowledge the Department of Genetics and Plant Breeding, Institute of Agricultural Sciences, Banaras Hindu University, for providing the HUM-27 (Malviya Jankranti) mung bean seeds and photographs. We also thank Ms. Vaishnavi (Department of Dairy Science and Food Technology) and Ms. Sushma Shrivastava (Department of Biochemistry) for their assistance with the analytical work.
 
Disclaimers
 
The results and conclusions are based solely on the authors’ experimental data and do not necessarily represent the views of the affiliated institutions. The authors are responsible for the accuracy and completeness of the information. Results may vary depending on mung bean variety and methodology.
 
Informed consent
 
The present study is part of PhD research and the study protocol was approved by the Institutional Ethics Committee (IAEC/6229), IMS, BHU.
 
Funding
 
Nil.

The authors declare that there are no conflicts of interest.