Materials and Methods:

1.1. Reagents: All Chemicals used were analytical grade quality. Nitric Acid (HNO 3) and
Hydrochloric Acid (HCl) were used to digest the samples. Stock standard solutions of
Copper, Iron, Magnesium, Manganese, Aluminum, Arsenic, Barium, Cadmium, Chromium,
and Zinc containing 100 ppm solutions in 2% HNO3 from HPS.

1.2. Collection of Shrimp Samples: There were varieties of shrimp samples collected from
the local grocery shops around Clinton, MS. The main species of interest were the collected
from the Gulf of Mexico, Argentina, Indonesia, and America. The samples were collected
from random shops to get collective data. Collected shrimp samples were kept in a deep
freezer to maintain their freshness and prevent rotting.

1.3. Sample Preparation for Analysis: The shrimp samples were in a frozen state. The shells
were removed and flesh from abdomen were cut and weighed approximately for 5 grams for
each shrimp and taken in sterilized plastic sample collecting tubes. To avoid contamination
of samples, certain measures were taken while sampling. They were marked as per the
location these samples were collected. This was followed by the addition of 5 ml of HNO 3
and 2 ml of HCl for digestion of the edible portion of the shrimps. Further, 3 ml of H2O2 was
added before these tubes were made to stay in Heatblock.

1.4. Heatblock Digestion: The closed vessel Heatblock digestion system (Environmental
Express heatblock-150) was used to digest the samples in collecting tubes. The sealed tubes
were kept in a heatblock and heated accordingly at a temperature of 95°C for a time of 2
hours and a cooling time of 15 minutes. After digestion was completed, the sample solutions
were cooled at room temperature and then filtered in new collecting tubes. Then the solutions
were made up to 30 ml by the addition of 5% (v/v) HNO3

1.5. Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES): Absorption

wavelengths of heavy elements like Copper, Iron, Magnesium, Manganese, Aluminum,
Arsenic, Barium, Cadmium, Chromium, and Zinc were detected using Agilant ICP-OES for
the presence of these elements in shrimp samples.