Triglycerides (TG) are lipid compounds consisting of a glycerol (a 3-carbon molecule containing three hydroxyl groups) backbone esterified to three long chain fatty acids. Triglycerides are a vital energy source for cells. Circulation of triglycerides through the vasculature is achieved by the incorporation of these compounds into lipoproteins. Lipoproteins are composed of a coat of phospholipid, cholesterol and proteins (apolipoproteins) enclosing a hydrophobic center of cholesterol esters and triglycerides. There are four classes of lipoproteins, high-density lipoproteins (HDL), low-density lipoproteins (LDL), very low-density lipoproteins (VLDL) and chylomicrons (CM). All these lipoprotein classes contain TG, however the highest triglyceride concentrations are found in CM and VLDL. Lipoproteins shuttle triglycerides from intestines (ingested triglycerides or CM) and liver (synthesized triglycerides, TG) to cells for energy or adipose tissue for storage. They also shuttle cholesterol to tissues (LDL) and liver (HDL) for use.




The concentration of triglycerides-rich lipoproteins (CM, VLDL, and IDL) in blood is controlled by a variety of hormones. Insulin, insulin antagonists, and other hormones affect lipoprotein concentrations by activating or inhibiting the activities of lipoprotein lipase, hepatic lipase, and hormone-sensitive triglyceride lipase. These hormone regulated enzymes stimulate metabolic processes such as lipolysis and hepatocyte clearance of lipoprotein remnants. Disorders disrupting CM and VLDL metabolism result in hyperlipoproteinemia (↑ [lipoprotein]) thus leading to hypertriglyceridemia (↑ [triglyceride]). Mechanisms of the enzymes involved in lipoprotein metabolism are described below:

  • Lipoprotein lipase (LPL): Lipoprotein lipase is found in vascular endothelium. It is activated by insulin, ACTH, TSH, glucagon and thyroid hormone. Its activity is enhanced by heparin. Lipoprotein lipase hydrolyzes CM and VLDL to free (non-esterified) fatty acids and glycerol for tissue use. Cholesterol-rich remnants produced from both lipoproteins are removed by the liver. Lipoprotein lipase is one of the several enzymes that also converts VLDL to LDL (via IDL). Apolipoprotein C-II is essential for activation of LPL.
  • Pancreatic lipase: Pancreatic lipase degrades dietary fat, allowing uptake and packaging into CM.
  • Hepatic lipase: This enzyme hydrolyzes surface phospholipids on lipoproteins and is responsible for removing triglycerides from LDL (and helps the liver remove LDL from circulation).
  • Hormone-sensitive triglyceride lipase: This enzyme is responsible for lipolysis (mobilization of triglycerides from adipose tissue to yield non-esterified fatty acids and glycerol). The enzyme is stimulated by catecholamines, glucagon, growth hormone, thyroxine, ACTH, corticosteroids and prostaglandins. It is inhibited by insulin (or the hormone is most active in the absence of insulin). Liberated non-esterified fatty acids are transported to the liver (free or albumin-bound), where they are taken up and used for energy (β-oxidation), re-combined with glycerol to form VLDL or incorporated into ketones. Therefore, lipolysis will increase VLDL production.


Reaction type

End-point reaction


  • Triglycerides GPO-PAP method: In the first step of this 4-stage reaction, the enzyme lipoprotein lipase (LPL) catalyzes the complete hydrolysis of triglycerides yielding glycerol and fatty acids. Step 2 involves the enzyme glycerokinase (GK) which, in the presence of Mg2+, catalyzes the phosporylation of glycerol from ATP. In the following step, glycerophosphate oxidase (GPO) catalyzes the oxidation of glycerol-3-phosphate, produced in the subsequent reaction, to dihydroxyacetone phosphate and hydrogen peroxide. Finally, under the catalytic action of peroxidase hydrogen peroxide reacts with 4-aminophenazone and 4-cholorophenol to form the red dye product 4-(p-benzoquinone-monoimino)-phenazone. The color intensity of the red dye product is measured photometrically, at 500 nm, and is directly proportional to the triglyceride concentration (reported in mg/dL).
  • Reactions are shown below:

triglycerides + 3H2O     lipoprotein lipase   > glycerol + fatty acids

glycerol + ATP    glycerol kinase  > glycerol-3-phosphate + ADP

glycerol-3-phosphate + O2 glycerol-3-phosphate oxidase> dihydroxyacetone phosphate + H2O2

H2O2 + 4-aminophenazone + 4-chlorophenol   peroxidase > red-colored complex

Units of measurement

Serum triglyceride concentration is measured in mg/dL (conventional units) or mmol/L (SI units). The conversion formula is shown below:

mg/dL x 0.1129 = mmol/L 

Sample considerations

Sample type

Serum, plasma, body cavity fluids (peritoneal, pleural)


Heparin or EDTA


Triglyceride concentrations in serum are stable for variable periods of time within a range of cold temperatures in humans: 10 days at 2-8 °C, 3 months frozen at -20°C, and several years frozen at -70°C (per product information sheet).  To maintain the highest attainable level of stability avoid repeated thawing and freezing cycles.


  • Lipemia : None of these interferents affect the triglyceride concentration substantially with the methods used by Cornell University. Note, that the term (result) lipemic index implies it is an assessment of triglyceride concentrations, the lipemic index in reality does not correlate well to triglyceride concentrations and is more of a reflection of sample turbidity.
  • Icterus: Can falsely decrease concentrations (icteric index > 9).
  • Hemolysis: No interference up to a hemolytic index of 700 (marked hemolysis).
  • Drugs: No interference was observed using common drug panels. Corticosteroids may increase triglyceride concentrations (resulting in a post-prandial lipemia)

Test interpretation

Low triglyceride concentrations are not of diagnostic relevance.

Increased concentration (hypertriglyceridemia)


Gross lipemia

Hypertriglyceridemia is due to increased CM and/or VLDL and may result in visible lipemia. It is important to remember that hypertriglyceridemia is often found in post-prandial samples. A fasting hypertriglyceridemia (lipemia) indicates underlying defects in lipoprotein metabolism. Lipemia should clear within 4-6 hours after eating, illustrating the importance of collecting a fasting blood sample for clinical pathologic testing (12 hour fast recommended). In dogs, the most common causes of a fasting lipemia are diabetes mellitus, hyperadrenocorticism, pancreatitis and corticosteroid therapy. In horses, hypertriglyceridemia is usually due to metabolic syndrome, pituitary pars intermedia dysfunction (PPID or equine Cushing’s disease), and negative energy balance (ponies, donkeys). Visible lipemia is usually only seen in states of negative energy balance in ponies and donkeys and is not a feature of equine metabolic syndrome or Cushing’s disease, despite hypertriglyceridemia in these conditions. In camelids, negative energy balance (with associated hepatic lipidosis) is a cause of hypertriglyceridemia and lipemia. In contrast, high triglyceride concentrations and visible lipemia are not seen in cattle with negative energy balance. This may be because cattle do not produce VLDL in response to lipolysis, but produce ketones (such as β-hydroxybutyrate) instead.

  • Physiologic: Post-prandial hypertriglyceridemia is due to an increase in CM.
  • Iatrogenic: Exogenous immunosuppressive or anti-inflammatory doses of corticosteroids can result in a fasting hypertriglyceridemia in dogs and cats (Lowe et al 2008, Khelik et al 2019, Tinklenberg et al 2020). This is attributed to inhibition of insulin action on lipoprotein lipase and stimulation of hormone-sensitive lipase. The increase will be due to CM and VLDL (lipoprotein lipase clears both from blood).
  • Pathophysiologic
    • Diabetes mellitus: Hyperlipemia is due to a marked increase in triglycerides, with a concurrent mild to moderate increase in cholesterol. There is increased CM, VLDL and LDL. This is from a deficiency or lack of insulin, resulting in decreased activation of lipoprotein lipase (decreases CM and VLDL hydrolysis), increased lipolysis (enhances VLDL production) and downregulation of LDL receptors (increases LDL). Hyperlipemia associated with diabetes mellitus (from paramyxovirus-mediated pancreatic necrosis) has been reported in llamas in the USA.
    • Hyperadrenocorticism: Corticosteroids inhibit the action of insulin, resulting in decreased activity of lipoprotein lipase (clears VLDL and CM). Corticosteroids also promote lipolysis by stimulating hormone-sensitive lipase, which will increase VLDL production.
    • Pancreatitis: Hyperlipemia is mostly associated with hypertriglyceridemia with a mild increase in cholesterol. This is due to increased CM and VLDL from decreased lipoprotein lipase activity.
    • Excessive negative energy balance: In states of excessive negative energy balance (e.g. starvation, anorexia) particularly when energy demands are high (e.g. late pregnancy, early lactation), lipolysis of fat stores in adipocytes will increase VLDL concentrations. Hyperlipemia due to excessive negative energy balance mostly occurs in horses and camelids and is due to increased energy demands from lactation or pregnancy combined with insufficient food intake (stress, transport, underlying disease), insufficient dietary energy or insulin resistance (from pregnancy or stress-associated hormones). In contrast, ruminants with excessive negative energy balance rarely develop triglyceride or cholesterol abnormalities (which has been attributed to inefficient export of VLDL by the liver in these species). In all species, excessive negative energy balance can cause hepatic lipidosis (non-esterified fatty acids from lipolysis are converted to triglycerides and are stored, when in excess, in hepatocytes).
      • Hyperlipemia in horses: This is especially seen in pony mares and donkeys and is associated with obesity, pregnancy, stress (e.g. transport) and lactation. It is characterized by negative energy balance (resulting in lipolysis) and insulin resistance (from pregnancy-associated hormones like progesterone and obesity). Horses have poorly developed pathways for ketone production and hence cannot convert mobilized fatty acids to ketone bodies, shifting them instead to VLDL production. Hypertriglyceridemia is due to increased VLDL and results in hepatic lipidosis (and eventually liver failure with increased liver enzymes), hypoglycemia, renal disease and central nervous system signs. It is fatal in more than 60% of cases. Common diseases associated with hyperlipemia in horses are hyperadrenocorticism (hyperglycemia, insulin resistance, and hirsutism are characteristic), colic, laminitis and gastrointestinal parasitism.
      • Hyperlipemia in camelids: Camelids, unlike horses, can also suffer from ketosis when they are in excess negative energy balance. Pregnancy toxemia is the term used to refer to pregnant camelids with hyperlipemia (due to increases in triglycerides [mostly VLDL] especially, but also cholesterol) and ketosis. Like horses, affected camelids often have underlying hepatic lipidosis and can die from their disease
    • Other metabolic conditions
      • Hepatic lipidosis: Anorexia induces increased lipolysis and hypertriglyceridemia (from increased VLDL production).
      • Hypothyroidism, cholestasis: Hypercholesterolemia is more common in these disorders, although triglycerides may be mildly increased (from increased VLDL production).
      • Familial hyperlipidemias: These are usually due to defects in lipoprotein lipase and have been reported in dogs (Miniature Schnauzers, Beagles, Brittany Spaniels) and cats (inherited hyperchylomicronemia in Siamese, domestic shorthair and Himalayans).
        • Hypertriglyceridemia in Miniature Schnauzers is seen in middle aged or older dogs (> 4 years old). They have marked hypertriglyceridemia (triglycerides of 500-800 mg/dL) with increased VLDL and CM. Many have concurrent diabetes mellitus or pancreatitis.
        • Inherited hyperchylomicronemia affects young cats (8-9 months old) and is associated with lethargy, inappetance, anemia, xanthomata (lipid granulomas in the skin and internal organs), neuropathy, and, if triglyceride concentrations are > 1500 mg/dL, lipemia retinalis. The disease is due to an autosomal recessive defect in lipoprotein lipase.