**6. Chemical tests of isolated biomaterial**

### **6.1 Tests for carbohydrate**

1 ml of freshly prepared biomaterial solution (5% w/v prepared biomaterial solution in double-distilled water) was taken in the test tube. Add two drops of Molisch reagent. Add 1–2 ml of conc. Sulfuric acid in the test tube and observed for the appearance of purple color at the interface of two layers formed. The test was performed and reported [4, 5, 40, 56, 63–65].

#### **6.2 Test for protein**

For testing the presence of protein in the isolated biomaterial was treated with 0.1% solution of ninhydrin reagent and 10% tannic acid solution. The presence of blue color and yellow color precipitate indicates the presence of protein. The test was performed and reported. Madhav and Yadav et al. [4] reported about the proteinous nature of biomaterials.

Biuret test was performed for the confirmation of proteins. 2 ml of *Prunus amygdalus* biomaterial was taken in the test tube (5% biopolymer solution in distilled water), add 1 ml of sodium hydroxide solution with the addition of copper sulfate solution drops. The mixture was kept aside for five minutes and observe any color changes. The appearance of violet color confirms the presence of proteins [37]. The test was performed and reported [4, 5].

#### **6.3 Spectral analysis of the biomaterial**

Spectral analysis of the isolated biomaterial was conducted like I.R, NMR, Mass spectroscopy, SEM studies. The biomaterials were subjected to IR, NMR, Mass spectroscopy studies, and the obtained spectra were interpreted and reported. SEM studies of different biomaterials were also performed and the obtained results were interpreted and reported. I.R, N.M.R, Mass spectroscopy was studied at Central Drug Research Institute, Lucknow, and SEM studies were studied at Birbal Sahani Institute of Paleobotany, Lucknow [62].

#### **6.4 SEM (Scanning electron microscopy)**

The isolated biomaterial's surface morphologies were characterized by Scanning Electron Microscope. In SEM analysis the external surface and internal structure were characterized. **Figure 1**. reveals about the flaky and rough surface of biopolymer [4].

**Figure 1.** *SEM of isolated biomaterial Litchi chinesis 1500x.*
