The opium latex of 1470 individual plant samples of 98 germplasm lines were screened for five major economic alkaloids. The alkaloids profile showed that morphine content ranged from 9.20–20.86% with an average of 15.00 ± 0.267. Codeine ranged from 1.69–6.48% with arithmetic mean of 3.35 ± 0.091, thebaine from 0.52–7.95% with an average of 2.27 ± 0.116. Narcotine ranged from 8.79–17.97% with an average 8.79 ± 0.266 and papaverine content ranged from 0.00–6.07% with an average 1.18 ± 0.167 in over all the germplasm lines. The content of different alkaloids categorized into class interval which exhibited that maximum number of plants and accessions for morphine content fall in group of 10–15% followed by 15–20%, for codeine content in group of 2–4% followed by 4–6%, for thebaine content in 1–2% followed by 2–4%, for narcotine in 5–10% followed by 10–15% and for papaverine content 0–2%. 24 germplasm lines had morphine content above 16.0%. Similarly, 37 germplasm lines for codeine and 13 for thebaine showed consistently high content, i.e., above 3%. 33 germplasm lines had narcotine content above 9% and 20 lines exhibited papaverine content above 2%. Line BR 316 was found unique having thebaine content 6.27–9.33%. The probable reasons of increase or decrease of different alkaloids based on their biosynthetic pathway has been discussed in detail.