# Spelt Grass Juice: Phytochemicals and Antiproliferative Activity
## 3 Results and Discussion  
### Photosynthetic Pigments in Extracts  
Various types of plant pigments are found in the photosynthetic apparatus as crucial components. By absorbing light of different wavelengths and allowing photosynthetic organisms to adapt permanently or temporarily to different environments, chlorophyll plays an important role in light harvesting. Among their diverse functions in photosynthesis, including light capture, carotenoids are essential antioxidants that lessen photodamage and photoinhibition [22].  
In green plants, as the two main chemical structures of chlorophyll are Chl a and Chl b, typically in a 3:1.1 ratio. Chlorophyll has proven to be an anticancer agent through multiple biological activities like antigenotoxicity, the trapping of mutagens, antioxidant activities, apoptosis, and immunomodulation [23]. Some limitations of its pure natural form are related to its poor stabilization under physiological conditions, and its poor solubility in aqueous solutions, which reduces its accumulation in cancer cells [24].  
The lyophilized samples of spelt grass juice (extracted with acetone-water) without foam had an average content of pigments as follows: 6.39 mg chlorophyll a/g DW, 2.47 mg chlorophyll b/g DW, and 1.81 mg carotenoids/g DW. Obviously, there were no significant differences between them and ones with foam: 6.91 mg chlorophyll a/g DW, 2.69 mg chlorophyll b/g DW, and 1.95 mg carotenoids/g DW (Figure 1, statistics not displayed). Methanol is known to be less efficient than acetone in the extraction of pigments. Consequently, the values for the mentioned pigments in the methanol-water extracts were about 5, 3, and 20 times lower than in to the acetone-water extracts, respectively.  
[PERSON] et al. [2] chromatography determined the content of photopigments in spelt grass extracts using pure acetone. The concentrations varied significantly, such as chlorophyll a 2.4-7.08 mg/100 g and chlorophyll b 8.55-28.1 mg/100 g of fresh spelt grass extract. These authors assume that significant amounts of chlorophyll a are converted into phenophytin a during the lyophilization process, for which they measured concentrations of 3.72-8.81 mg/100 g. Further, the sum of average values for carotenoids (\(\alpha\)-carotene, \(\beta\)-carotene, lutein, and zeaxanthin) ranged from 2.45 to 3.08 mg/100 g. Our results showed a much higher content of chlorophyll a (about 40 mg/100 g FW) and carotenoids (more than 10 mg/100 g FW), while the content of chlorophyll b (about 15 mg/100 g FW) is in the range of values found in the study of [PERSON] et al. [2].  
[PERSON] et al. [8] measured the content of total chlorophyll pigments in wheatgrass juice 481 mg/L for the summer and 270 mg/L for the winter period with a chlorophyll a/chlorophyll b ratio of 1:0.34 and 1:0.27, respectively. For carotenoids, the concentrations were 94.3 mg/L in summer and 59.4 mg/L in winter. In our experiment with young spelt grass grown under controlled conditions, the total chlorophyll content was about 550 mg/L with a chlorophyll-a-to-b ratio of 1:0.38, and the carotenoid content was about 110 mg/L, which could be considered quite similar to their results for summer wheatgrass juice. It is important to note that [PERSON] et al. [8] also used 80% acetone for the extraction, and a spectrophotometric method to determine the pigment concentration.