The effect of time of day, fasting and meal type on biomarkers of liver injury in healthy subjects

INTERVENTION: Each healthy volunteer received three different diets in one of four possible sequences (randomized 1:1:1:1 using a computer generated randomization schedule) each for 8 days, which was expected to be a sufficient duration to detect liver enzyme changes. The four possible diets included (1) standard diet (3000 kcal, 55% carbohydrates, 30% fat,15% protein) (2) standard high‐calorie diet (4500 kcal, 55% carbohydrates, 30% fat, 15% protein) (3) high‐fat high‐calorie diet (4500 kcal, 25% carbohydrates, 60% fat, 15% protein) (4) high‐carbohydrate high‐calorie diet (4500 kcal, 65% carbohydrates, 20% fat, 15% protein) During each period, participants spent 10 days (11 nights) at the Clinical Research Unit. Meals were administered at intervals of approximately five hours, except on Day 9, when participants fasted until 1 pm. This was followed by a non‐resident washout of at least 11 days, during which participants consumed their usual diet. CONDITION: Effect of diet on markers of liver injury in healthy individuals ; Digestive System PRIMARY OUTCOME: ; 1. Hepatocellular injury biomarkers: ALT, AST, and glutamate dehydrogenase (GLDH) measured using blood test at baseline and Day 2, Day 3, Day 5, Day 8, Day 9 and Day 10; 2. Hepatobiliary injury biomarkers: ALP and gamma‐glutamyl transferase (GGT) measured using blood test at baseline and Day 2, Day 3, Day 5, Day 8, Day 9 and Day 10; 3. Hepatic function biomarkers:; 3.1. Total bilirubin measured using blood test at baseline and Day 2, Day 3, Day 5, Day 8, Day 9 and Day10; 3.2. Fasting total bile acids using blood test at baseline and Day 2 with hourly measurement from Hour 1 to Hour 16, Day 2, Day 3, Day 5, Day 8 with hourly measurement from Hour 1 to Hour 16, Day 9 with hourly measurements from Hour 1 to Hour 16, Day 10; 4. Total bile acids during the post‐meal period (8am‐1pm, 1pm‐6pm, 6pm‐8am) measured using blood test at Day 9 during the post‐meal period (from Hour 1 to Hour 6, Hour 6 to Hour 10, hour 10 to hour 24; 5. Concentration of novel biomarkers (GLDH, micro RNA) measured using blood test at baseline and D2, Day 3, Day 5, Day 8, Day 9 and Day 10.; SECONDARY OUTCOME: ; 1. Total cholesterol, low‐density lipoprotein (LDL), high‐density lipoprotein (HDL), and triglyceride measured using blood test at baseline and Day 2, Day 3, Day 5, Day 8, Day 9 and Day 10; 2. Total bile acids and triglycerides at baseline and hourly from 8 am ‐ 1 pm on Day 8 (fed) and Day 9 (fasted) measured using a blood test; 3. Concentrations of bilirubin, ALT, AST, GGT, ALP, total cholesterol, LDL, HDL, triglycerides, and creatine kinase (CK) measured using blood test at baseline and diet on Day 9 and Day 9 with fasting; INCLUSION CRITERIA: 1. Aged 18‐55 years old 2. No clinically relevant abnormalities identified by the medical screening 3. Body mass index (BMI) of 17.5‐30.5 kg/m² 4. Total body weight >50 kg 5. Smoked =5 cigarettes daily 6. Free from the use of prescription or non‐prescription drugs/dietary supplements for at least seven days or five half‐lives prior to the first study period