Cobas kits

The insulin and lipase levels were measured according to the guidelines on the supplier's standard kits using the Cobas® kits (Roche Diagnostics, USA). The insulin test was critical in examining the overall effects of P. dactylifera seeds as it directly measures the level of insulin in rats [31 (link)]. Lipase enzyme helps the body in breaking down the fats in the intestine. Any fluctuation in its level is considered an abnormality.

Biochemical markers of the liver (aspartate aminotransferase (AST), alanine aminotransferase (ALT) and total bilirubin) and kidney (blood urea nitrogen (BUN) and creatinine) of respective organs were estimated in the serum samples according to the supplier’s guidelines using Cobas® kits (Roche, USA).

Based on the values of UIC obtained and after exclusion of non-respondents, a systematic random sampling methodology was carried out and blood samples were taken from 104, 150, 104 and 61 students with normal, mild; moderate and sever UIC, respectively. The collected blood sample was incubated for an hour at room temperature and the unclotted portion of the blood was centrifuged at 2000 rpm for 20 minutes. The principle of the assay is Electrochemiluminescence (ELC) immuneassay, using Roche Cobas kits following manufacture’s instructions.12 (link) The normal reference ranges of total T4, T3 and total TSH were 4.5-12.6µg/dL, 0.39-1.37ng/dL and 0.3- 4mIU/L, respectively.13

Participants’ menstrual patterns and demographic characteristics, such as age, weight, height, etc., were recorded. BMI (weight (kg)/height (m)²) were calculated. BMI was categorized as normal (BMI 18.5–24.9), class-1 obese (BMI 30.0–34.9), and class-2 obese (BMI 35.0–39.9).
After eight hours of fasting, venous blood samples were drawn from a forearm vein during the menstrual period (Days 2–3) in the morning. All blood samples were centrifuged to obtain serum within 30 minutes after bloodletting. Serum samples were then kept at -80°C until GDF-9, BMP-15, kisspeptin, AMH, FSH, LH, estradiol, glucose, and insulin serum level analysis were performed. The homeostatic model assessment of insulin resistance index (HOMAIR) was utilized to quantify insulin resistance = Fasting glucose (mmol/L) x Fasting serum insulin (μU)/L)/ 405.
Serum insulin, glucose, FSH, LH, and estradiol levels were measured using the electrochemiluminescence immunoassay technique with the Cobas 8000 analyzer using Roche Cobas kits (Roche Diagnostics GmbH, Mannheim, Germany). The quantitative ELISA kits protocol used to measure the main peptides are shown in Table 1.