*4.7. Statistical Analysis*

All of the data were made in duplicate and are expressed as the means of the measurements ± standard deviation. The one-way analysis of variance (ANOVA) with Tukey's test were made to test the differences between means at a 5% significance level by using the XLSTAT statistical package (2021 version, Addinsoft, Inc., Brooklyn, NY, USA).

#### **5. Conclusions**

The germination period significantly influenced the growth of the seeds, leading gradually to development of the radicle and first leaves. The seed development was captured by a stereo microscope device which showed that a four-day germination may be optimum for legumes used in food consumption. The microstructures of the seeds during germination changed, ascribed in the case of bean, chickpea and lentil mainly to the starch, and in the case of lupine and soybean to the protein. Germination of seeds resulted in increased protein and ash contents for lentil and chickpea, whereas for the rest of the legume seeds their contents showed different variations. Additionally, during the germination period the fat content varied for bean and soybean, whereas it decreased for lentil, chickpea, and lupine. Mineral contents (sodium, magnesium, zinc, iron) of the germinated legume seeds increased during germination in all legume seeds, showing the beneficial influences of germination on the nutritional profile of legumes. An increase in acidity values and a decrease in pH and carbohydrate contents have also been recorded for all legume types during the germination period. Fourier transform infrared spectroscopy (FT-IR) highlighted variation in chemical compounds of legume seeds during the germination period. According to the wavelength and peak height of the FT-IR spectra, we clearly showed the fact that different compounds such as protein, carbohydrates and lipids varied depending on germination period and each legume type. Principal component analysis was performed on the combined physico-chemical and minerals data, clustering the legume types obtained during the germination process in the PC space. PCA highlighted an association between lentil, bean and chickpea samples which were placed in the right part of the graph and between soybean and lupine samples which were placed in the left part of the graph, indicating similar compositions of these samples. From the physical-chemical data point of view, significant negative correlations were obtained between carbohydrate contents and protein, fat and ash variables at a level of 0.01.

**Author Contributions:** D.A., S.-G.S. and G.G.C. contributed equally to the study design, collection of data, development of the sampling, analyses, interpretation of results, and preparation of the paper. All authors have read and agreed to the published version of the manuscript.

**Funding:** This research received no external funding.

**Institutional Review Board Statement:** Not applicable.

**Informed Consent Statement:** Not applicable.

**Data Availability Statement:** Not applicable.

**Acknowledgments:** This work was supported by a grant of the Romanian Ministry of Education and Research, CNCS—UEFISCDI, project number PN-III-P1-1.1-TE-2019-0892, within PNCDI III.

**Conflicts of Interest:** The authors declare no conflict of interest.
