Compositional profile of barley landlines grown in different regions of Gilgit‐Baltistan

Abstract The current investigation was performed to explore the nutritional and functional composition of four landlines of barley denoted as LB1 (Gilgit), LB2 (Nagar), LB3 (Skardu), and LB4 (Shigar) from different regions of Gilgit‐Baltistan. The samples were examined for nutritional profile and antioxidant attributes. Total phenolic values and total flavonoid results were in the range of 1.2 to 3.1 mg/g and 0.41 to 0.55 mg/g, respectively. Nutritional profile as crude starch, fiber, protein, ash, and fat ranged from 56.3%–50.80%, 16.50%–11.73%, 16.20%–11.53%, 2.8%–2.1%, and 2.63%–1.63%, respectively. The mineral composition in terms of Mg (527–616 mg/kg) was higher in the landlines followed by Ca (312–368 mg/kg), Na (122.6–146.6 mg/kg), Fe (43.3–65.6 mg/kg), and Zn (22.5–26.6 mg/kg). It was concluded that the indigenous barley landlines had immense nutritional potential and functional attributes. Thus, it can be used for value‐added food products and the development of cottage industry in the region.


| Samples collection and preparation
The current study was conducted in the Advanced Instrumental Laboratory of Karakorum International University. The samples of dried barley LB 1 (landline barley) Gilgit, LB 2 (Nagar), LB 3 (Skardu), and LB 4 (Shigar) were collected from the farmers in different districts of Gilgit-Baltistan, Pakistan. The collected samples were cleaned manually for foreign residues and other impurities. After that, the samples were grounded in flour (Mesh size) with the grinding mill, and the final product (weight) was stored in polythene bags for further analysis under ambient conditions.

| Free radical scavenging activity
The total antioxidant characteristics of all the samples were detected by following DPPH (2, 2-diphenyl-l picryl hydrazyl) technique reported by (Mareček et al., 2017) with minor changes. In detail, DPPH solution was prepared and then followed by covering with aluminum foil and stored under refrigeration temperature for its further use. For antioxidant estimations, 5 g of each barley flour sample was homogenized and then extracted by using methanol (10 ml) for 48 hr. Further, in a volumetric flask (100 ml volume), 0.1 ml extract and 3.9 ml of DPPH solution having 6 × 10 -5 mol/L concentration was mixed and then incubated at ambient temperature for 35 min. After the incubation time, the absorbance was calculated at 517 nm with UV spectrophotometer. The antioxidant attributes were estimated by employing the expression as under:

| Total phenolic content
Total phenolic values were recorded by following the Folin-Ciocalteu procedure, as described by Shahzad et al. (2020) with minor modifications. Briefly, 5 g of the barley sample was first ground into powder followed by homogenization and then extracted by using 10 ml methanol for 48 hr. Further, 1 ml (mg/ml) extract was mixed gently with 4.6 ml distilled water and 1 ml Folin-Ciocalteu (1N). After 3 min, 3 ml sodium carbonate (2%) was mixed into the mixture and stand it for 2 hr. Finally, the absorbance was recorded at 760 nm by using a UV spectrophotometer.

| Total flavonoid content
The total flavonoid values of the barley samples were tested by according the method described by Manzoor et al. (2019) with minor modification. 1 ml of the samples extract (1 mg/ml) was taken and mixed with 4ml distilled water. Further, 0.3 ml AlCl 3 (10%) and 2ml of the NaOH (1N) were also poured into the reaction flask. Again, 2.7 ml of distilled water was added, agitated well, and then, absorbance was recorded at 510 nm. Various concentrations of quercetin were employed as an internal standard.

| Moisture
The moisture content of barley flour was examined by using the protocol set by AACC (2000)

| Crude fat
For crude fat determination, dried samples were processed in the soxhlet method. In which, continuous refluxing was done by using petroleum ether as solvent as reported by AACC (2000) Method No.
30-10.01. In detail, a 3 g sample was weighed and dried in an oven till constant weight. The dried sample was then wrapped in filter paper and put in soxhlet apparatus and 5 to 6-time washings were given with petroleum ether as extraction solvent. The solvent was evaporated after extraction, and fat content was determined by employing the formula mentioned below.

| Crude ash
For Ash content analysis, AOAC (2006) Method No.923.03 was followed. Firstly, ignited the empty crucibles at 550°C, weighed and, then, cooled in a desiccator to room temperature.
Then, took a 2 g homogeneous sample in a crucible and placed it in a muffle furnace at 660°C until light gray mass was achieved.
Finally, the crucibles were removed from the furnace and allowed to cool down in a desiccator. Calculate the weight of ash along with the crucible and calculate the net weight. Ash content was recorded by using the formula as under; whereas; W 1 = crucible weight; W 2 = sample weight; W 3 = sample weight after ashing.

| Mineral contents
The mineral content of the barley flour was ascertained by employing the wet digestion method proposed by AOAC (2006). For which, 0.5 g premixed sample was first digested at 60-70°C, by using HNO 3

| Statistical analysis
All measurements were carried out in triplicates, and it was analyzed with the help of statistics 8.1 (Tallahassee FL 32,317, USA). Oneway analysis of variance (ANOVA) was applied in factorial design at p <.05 choose as significant.

| Antioxidant activity of landline barley samples
The DPPH radical scavenging activity of landline barleys from different districts is presented in

| Total phenolic and flavonoids components of barley samples
The total phenolic contents of landline barley samples from different districts are presented in Table 1. The findings for the tested parameters from different districts were significantly different from each other (p <.05). The total flavonoid content was estimated highest in LB 2 (3.1 mg/g) followed by LB 3 (2.9 mg/g), LB 1 (1.9 mg/g), and LB 4 (1.2 mg/g).
The findings of the current study were quite similar to the study conducted by Abidi et al.(2015), which reported 47-123 mg CE/100 g.
Similarly, our findings were slightly higher than those of Bellucci et al. (2013), calculated as 26.9 mg/100g in Dutch barley. The quantity and quality of polyphenols may be affected by some factors such as plant genetics and cultivar, soil type, growing methods, maturity stage, and postharvest management (Taranto et al., 2017). Flavonoid content in barley changes according to variety; white, blue, and purple kernels have a high concentration of flavonoid among others (Liu et al., 2013).
The results about total flavonoid contents (TFC) among different landlines are depicted in

| 3 Nutritional composition
The chemical composition of landline barley samples in different districts is depicted in  Moreover, the results regarding ash content showed significant differences (p <.05) among the tested samples. The highest ash content was analyzed in LB 2 (2.86%) followed by LB 4 (2.70%), LB 3 (2.43%), and LB 1 (2.1%), whereas the minimum value was recorded in LB 1 to be 2.1%. These findings were closely related to the results of Brennan and Cleary (2005). They assessed total ash content in whole grain barley ranging 1.5%-2.5%. Furthermore, our findings were also in line with a study conducted by Quinde-Axtell and Baik (2006). They determined ash content as 2%-3% in the barley samples.
The crude fat content for LB 4 was significantly lower than other samples. However, there was no significant difference between LB 1 , LB 2 , and LB 3 samples were recorded. Our findings were closely related to Brennan and Cleary (2005), who reported 2%-3% total lipids in barley, whereas Quinde-Axtell and Baik (2006)  LB 4 (50.8%). These changes might be due to genetic differences and cultivar (Wozniak et al., 2014). The nutritional composition of cereal grains might be affected by the environmental conditions under they grow and many studies have shown differences in concentration of fat, protein, and β-glucan content in oat and barley grown under different environment (Redaelli et al., 2013). Moreover, Ping et al. (2013)