Identification of Phytochemicals from Seed Extract of Custard Apple, Annona squamosa

Custard apple has many alkaloids, such as aporphine, roemerine, norcorydine, corydine, norisocorydine, glaucine and anonaine in different parts of the plant. The roots are used to treat acute dysentery, depression and spinal marrow diseases, while leaves have been used in cases of prolapse of the anus, sores and swelling. Ripe fruit is sweet and good tonic for human health and it is enriching the blood, increases muscular strength and lessens vomiting. Its seeds are used as abortifacients. Seed oil is used in paint and soap industry. This investigation involves preliminary screening, detection, and separation of secondary metabolites from the seed extract of Annona squamosa Linn. Oil was extracted from seeds of Annona squamosa Linn. by Soxhlet extraction method; methanol was used as a solvent for extraction. Absorbance and functional group detection were studied using FTIR, seed oil supernatant was used to detect the functional groups of secondary metabolites those were presented in sample. A potent Cyanidin-3-O-(2-O-beta-xylopyranosyl-6-O-acetyl)-beta-galactopyranoside, 4-[4-(2aminoethyl)-2,6Diiodophenoxy]2-iodophenol, Amikacin. The study was done using atmospheric pressure chemical ionization (APCI) Liquid Chromatography mass Spectroscopy technique for identification of Phytochemical. Various phytochemicals were detected using this technique. The analysis by APCI-LC-MS reveals presence of several compounds like cyclopeptides and acetogenins. Cyclosqamosin A, Cyclosqamosin B, Cyclosqamosin H, Acetogenins (polyketides); Annonacin, Squamocin, Annonin VI, which were detected by peaks of m/z ratio between 605 to 640 positive ions shift and Tenacissoside F (steroid) at 667 m/z negative ion shift. Identified compounds were compared with reference of earlier investigations, it was clear that this compound played a major role as anti-diabetic, anticancer, anti-inflammatory and have insecticidal property. However, further Research is required to study phytochemicals.


INTRODUCTION
Custard apple is tropical fruit which belongs to genus Annona and family Annonaceae and are collectively known as annonaceous fruits. There are over 120 species of genus Annona and are commonly found in India as a fruit consuming plant. This plant is commonly known as custard apple in english and Sitafal in gujarati. Custard apple contains alkaloids, such as aporphine, roemerine, norcorydine, corydine, norisocorydine, glaucine and anonaine in various parts of the plant (Kowalska and Puett, 1990;Pinto et al., 2005;Hiwale, 2015). The Custard apple roots are used in the treatment of acute dysentery, spinal marrow diseases and some cases of depression. The leaves of Custard apple are used in cases of prolapse of the anus, sores and swelling. Custard apple contains alkaloids, such as aporphine, roemerine, norcorydine, corydine, norisocorydine, glaucine and anonaine in various BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS parts of the plant (Kowalska and Puett, 1990;Chao-Ming et al., 1997;Pinto et al., 2005). Seeds of Custard apple contains acetogenins namely; squamocins B to n, coumarinoligans, annotemoyin-1, annotemoyin-2, squamocin and cholesteryl, glucopyranoside (Zahid et al., 2018).
Seeds of Custard apple are toxic, but they are used to treat head lice as they have insecticidal properties (its preparation causes eye irritation and can cause blindness). Seeds of Annona also have insecticidal properties. Farmers use pesticides to protect their crops from pest infestation, using chemical pesticides is no longer preferable. Seeds of custard apple can be used as biopesticide which can be used as suitable alternatives for pest control. Oil content is high in seeds which can be used to make soap or, if treated to remove the toxic alkaloids, it can be used as a cooking oil (Das et al., 2016;Vetal and Pardeshi, 2019). earlier studies have been done to study phytochemicals obtained from seeds, leaves.  , 2020). This investigation involves preliminary screening, detection, and separation of secondary metabolites from the seed extract of Annona squamosa Linn. By comparing with reference of earlier studies, it was clear that seed extract contains compound that played a major role as anti-diabetic, anticancer, anti-inflammatory and have insecticidal property (Mangal et al., 2015;Ribeiro et al., 2018). The present investigation involves identification of such compounds that can be used for medicinal purposes.

MATERIAL AND METHODS
Seeds of Annona squamosa were collected from fruit of Annona squamosa L. seeds were tested and determined viability, viable seeds were selected for identification of phytochemicals as previous study (Patel et al., 2019). Seeds were crushed using mixer grinder and powdered seeds were filled in a paper cup for further oil extraction. Finely crushed seed powder packed in paper cup was taken for oil extraction using Soxhlet extraction method, methanol was used as a solvent for extraction. After few cycles the crude oil was obtained and stored in dark bottles for further phytochemical analysis.
FTIR spectral analysis was conceded from the Annona squamosa seed oil supernatant to detect the functional groups of secondary metabolites those were presented in sample.  (Chavan, 2006).       4) and APCI (negative), we have obtained major peaks of m/z at 667, 683 (Fig 5). The analysis by APCI-LC-MS revealed presence of several compounds namely cyclopeptides and acetogenins (Zahid et al., 2013 (Mangal et al., 2015;Ribeiro et al., 2018).

CONCLUSION
The present study explicates the therapeutic application of the seeds which is a rich source of antioxidants such as phenols and flavonoids. The analysis by APCI-LC-MS and FTIR reveals presence of several compounds; cyclopeptides and acetogenins. Cyclopeptides like Cyclosqamosin A, Cyclosqamosin B, Cyclosqamosin H and other groups of cyclopeptides. Acetogenins (polyketides), Annonacin, Squamocin, Annonin VI and Tenacissoside F. By comparing with reference of earlier studies, it was clear that this compound played a major role as anti-bacterial, anti-diabetic, anticancer, anti-inflammatory and have insecticidal property. Research and development would be an important area to focus on medicinal importance of plant. The isolated compounds can be used in future to make antidiabetic, anticancerous, anti-inflammatory medicines, further in future this compound can be used in anti-leaching cream, anti-dandruff shampoo, hair oil and seed oil can also be used as biopesticide.

ACKNOWLEDgEMENTS
This study has been supported by Department of biotechnology, Pramukh swami science & H.D Patel Arts College, Kadi. We are highly thankful to principal Dr. Ajay S. gor, Lab Assistant Mayur Shah and Viral patel for constant technical support.

Conflict of interest:
On behalf of all authors, the corresponding author states that there is no conflict of interest.