*4.2.2 Exclusion criteria*


#### **4.3 Experimental procedure**

Recruited study subjects were non-randomised into two intervention groups: a lycopene group (n=10) and a placebo group (n=10). This non-randomised method was chosen to ascertain the similarity of subject characteristics between study groups with respect to age, gender distribution and total cholesterol. Because the experimental groups were small (n=8–10), randomisation would likely increase the risk of variable group composition.

The Principal Investigator was provided with coded study product, according to the number of subjects in intervention(s) and control, and allocated an equal number of study subjects to either study product such that each group (corresponding to the same product code) matches the other group in numbers and demographic and clinical variables (as explained above). The Principal Investigator was blinded to which codes represented active or control product.

Prior to the start of the study, subjects were instructed to refrain from the consumption of dairy butter and tomato-based products for 10 days before beginning the trial and for the duration of the study. At the start of the study, following the end of the 10-day run-in period, blood samples were drawn and subjects were checked for meeting inclusion criteria. Then, subjects were distributed with a 2-week supply of the study product. Anthropometric measurements and blood pressure were taken and blood samples drawn.

All volunteers were instructed to ingest one sample of dairy butter once a day with the main meal.

All blood parameters were measured in the morning between 8 and 10 am.

Blood was collected in the morning after an overnight fast from arm veins of the volunteers. Serum was separated from the clotted mass by centrifugation and aliquots were stored at –80°C prior to analysis.

#### **4.4 Compliance**

All volunteers were instructed to ingest dairy butter sample once a day with the main meal. All volunteers were instructed to keep their butter packaging and bring it to the following clinical visit. After the verification of compliance, the packaging was exchanged for a fresh 14-day supply of products.
