Seven standards were separated within 5 min in UPLC. In case of UFW, the main phenolic compounds detected in UPLC as well as in TLC short wave UV were 4-hydroxy benzoic acid (HBA; 0.26 mg/g wheat) and 4-hydroxy-3-methoxybenzoic acid (HMBA; JNJ-26481585 0.22 mg/g wheat), whereas, two other unknown compounds (UU1 and UU2) were detected in UPLC. According to TLC of ROFW sample, three bands were identified in short wave UV namely HBA, HMBA

and an unknown compound (SU), while, in long wave UV four unknown bands were observed (LU1-4). In UPLC profile of ROFW, HBA (1.61 mg/g wheat) and different unknown phenolic compounds (majorly RU1-5) were detected. Those unknown compounds might be contributing big role for the antioxidant property of ROFW. Through UPLC analysis [17], observed that syringic acid was the main compound (75.3–77%) in the free phenolic extracts (80% ethanol) of wheat meal. In our study, water was used as extracting solvent. Water extracts were phase separated by ethyl acetate, dried, dissolved in methanol and then they were injected in UPLC column. It is very difficult to compare our data with literature because different methods have been used for extraction. Moreover, antioxidant property

varies between species and varieties of grains [34]. SSF is a complex biochemical process where several hydrolyzing enzymes like α-amylase, xylanase, β-glucosidase, esterases, etc., are produced, which are predicted to be associated in the release of water soluble and more bioavailable PC from insoluble bound form [4]. Along with enzymatic release of PC, some other unknown biochemical pathways might be involved in SSF process to increase the TPC and antioxidant Raf targets properties of wheat. Moreover, in addition to PC, some other water soluble bioactive compounds like small peptides, xylo-oligosaccharides etc., produced during SSF might be contributing in the enhancement of antioxidant properties of fermented wheat [19], [9] and [28]. Present study demonstrates that SSF of wheat by R. oryzae RCK2012 is a very fruitful method for the enhancement of TPC and antioxidant potential. On the basis of the results obtained, it is clearly indicated that water extract of ROFW has strong antioxidant property Reverse transcriptase against

several in vitro and an in vivo oxidative system compared to unfermented wheat. Predominantly insoluble bound phenolics in cereals are less bioavailable. To maximize the possible health benefits of cereals, SSF is a great option for the improvement of bioavailability of cereal phenolics by increasing their solubility. In comparison to unfermented wheat, consumption of fermented wheat might give more health protection against oxidative damages. Moreover, fermented extract can be served as powerful sources of natural antioxidants over the synthetic antioxidant compounds used very often in food and pharmaceutical industry. Additionally, along with PCs some other bioactive compounds might be produced during SSF, which were contributing antioxidant property.