Does the composition of breast milk change over the course of the 24-hour day? Yes. The "drowsy" hormone, melatonin, reaches peak concentrations in breast milk at night. Cortisol -- a stress hormone that promotes alertness -- is typically at its highest in the morning.
So if you feed your baby milk that has been pumped and stored, it's probably a good idea to make sure the milk is "circadian-matched." Researchers suspect that "mistimed" milk could contribute to sleep troubles, and possibly disrupt your baby's developing circadian rhythms.
Some researchers call it "chrononutrition," the idea that we can improve our sleep cycles and other daily rhythms by adjusting the timing of what we eat.
For example, some food ingredients make us more alert. Others make us sleepy. And breast milk? It might be able to do both, depending on when it's produced.
So maybe it will become standard operating procedure. Mothers who use breast pumps and store their breast milk will note the time of day that their milk was expressed. When they feed their babies at night, they’ll use only milk that was produced after dark.
And when parents use baby formulas? Maybe they'll use slightly different formulations depending on the time of day -- to mimic the natural changes in breast milk.
Consider, for example, a study of five breastfeeding mothers in Israel.
Researchers tracked changes in breast milk over time, testing milk every 2 hours for a 24-hour period. In particular, the researchers measured levels of melatonin, the hormone that makes us feel drowsy after dark.
Were there any differences between milk expressed during the day and milk expressed in the evening?
Yes, there were, and the researchers found evidence of a clear, circadian rhythm: Milk produced at night contained higher concentrations of melatonin (Cohen Engler et al 2012).
Subsequent research has confirmed these results.
For instance, in a study of 21 lactating mothers living in Germany, researchers didn't just find that melatonin levels were higher at night. They were 5 times higher than daytime levels (Katzer et al 2016).
Similarly, researchers in China have documented dramatic changes across a 24-hour period.
Yishi Qin and her team tested the breast milk of 98 lactating mothers, and compared milk melatonin levels at 3pm, 9pm., and 3am (Qin et al 2019).
On average, melatonin levels at 9pm were nearly 3 times as high as melatonin levels at 3am. And the breast milk pumped in the middle of the night? At 3am?
That's when melatonin concentrations were at their peak -- nearly 10 times higher than melatonin levels in milk collected during the afternoon.
Cortisol helps make us feel alert, and it's possible that babies could receive a boost in cortisol depending on the timing of breast milk production.
Morning levels are approximately 4 times higher than levels present in breast milk produced in the evening (around 6pm). And they are about twice as high as levels present in milk expressed during the night (Pundir et al 2017; Italianer et al 2020).
Tryptophan is found in breast milk, and it is used by the body to manufacture melatonin. Like melatonin, tryptophan levels rise and fall according to a circadian rhythm, with concentrations peaking at night.
Do babies who consume tryptophan in breast milk experience a rise in melatonin levels? There is some evidence to support the idea.
Javier Cubero and colleagues studied 8 exclusively-breastfed infants, tracking changes in their mothers’ breast milk and changes in the infants’ melatonin levels. They also monitored a control group of 8 formula-fed babies (Cubero et al 2005).
All the babies were approximately 12 weeks old, and they were each fed on the same schedule -- once every 4 hours. But there was a difference between groups. For breastfed babies, melatonin levels peaked a few hours after their mothers’ tryptophan levels did. For formula-fed babies, the peak in melatonin happened much later, and infants spent significantly less time sleeping at night.
Did the tryptophan in breast milk cause differences in melatonin rhythms and sleep patterns?
That’s not clear. Possibly, the link between maternal tryptophan and infant sleep patterns was due to chance. And even if it wasn’t, we can’t be sure about causation.
Maybe something other than maternal tryptophan could explain why breastfed babies were different. For example, breastfed babies might have gotten more skin-to-skin contact, which lowered their stress levels and made it easier to sleep. Or maybe the breastfeeding mothers in this study were more likely to expose their babies to natural lighting patterns, which would have helped their babies adjust to the 24-hour day.
So I’m interested in a study conducted by Sara Aparicio and colleagues (Aparicio et al 2007). These researchers tested the effects of dietary tryptophan by performing a double-blind experiment on formula-fed babies (Aparicio et al 2007).
There were three conditions:
After a week on the “night-time tryptophan” regimen, babies in the test condition showed improvements in their sleep patterns. Babies in the control groups did not.
So it seems that higher daytime tryptophan concentrations might help young babies sleep better.
Melatonin and tryptophan aren't the only sleep-associated substances in breast milk. Cristina Sanchez and her colleagues note that several nucleotides—like 5’UMP, 5’AMP, and 5’GMP—either induce sleepiness or help regulate circadian rhythms (Sanchez et al 2009).
These nucleotides are present in breast milk. Do their concentrations in breast milk change over a 24-hour period? The researchers asked 30 women to express their breast milk at several different times of day. Then the milk was analyzed.
Nucleotide concentrations in breast milk did indeed change over a 24-hour period.
For example, 5’UMP--which has a calming effect--peaked in the middle of the night.
And 5’AMP, which makes people feel drowsy, was at its highest concentration in breast milk that was expressed in the early night-time.
Can we assume that nucleotides in breast milk might help infants fall asleep—and stay asleep—during the night?
To answer this, we need another experiment like that one conducted on formula-fed babies by Sara Aparicio’s team. Some experiments have been conducted, but they involved food that was fortified with both tryptophan and sleep-friendly nucleotides (Cubero 2006; Cubero et al 2007). As a result, we don’t know what effect the nucleotides might have had on their own.
More research is needed to answer this question, and the reality is bound to be complex. But as Jennifer Hahn-Holbrook and her colleagues have noted, it's certainly plausible.
Animal studies confirm that the melatonin in breast milk "rapidly crosses the intestinal barrier and diffuses into many tissues, including the liver, kidney and brain" (Hahn-Holbrook et al 2019).
It's also clear that glucocorticoids (like cortisol) can pass from breast milk into a young rodent's brain, and we've good reason to think that the same is true for humans (Hahn-Holbrook et al 2019).
So it seems wise to assume that the timing of breast milk production could make a difference. It may have an impact on an infant's behavior, and not merely in the minutes or hours after a baby feeds.
Hahn-Holbrook and her colleagues point out that a regular diet of "mistimed milk" might interfere the development of a baby's circadian rhythms, contributing to sleep problems. It might also alter an infant's stress responses.
The good news? If mistimed milk really does present problems, these are easy to fix. We just need to note the time of day when milk is expressed, and avoid giving our babies milk that is chronologically mismatched.
For more information about the composition of breast milk, click here.
For more breastfeeding information, including tips about breast pumps, see these practical breastfeeding tips.
Struggling with a sleepless baby? Check out these evidence-based articles about the science of baby sleep.
Aparicio S, Garau C, Esteban S, Nicolau MC, Rivero M, and Rial RV.
2007. Chrononutrition: use of dissociated day/night infant milk
formulas to improve the development of the wake-sleep rhythms. Effects
of tryptophan. Nutr Neurosci. 2007 Jun-Aug;10(3-4):137-43.
Cohen Engler A, Hadash A, Shehadeh N, Pillar G. 2012. Breastfeeding may improve nocturnal sleep and reduce infantile colic: potential role of breast milk melatonin. Eur J Pediatr. 171(4):729-32.
Cubero J, Valero V, Sánchez J, Rivero M, Parvez H, Rodríguez AB, Barriga C. 2005. The circadian rhythm of tryptophan in breast milk affects the rhythms of 6-sulfatoxymelatonin and sleep in newborn. Neuro Endocrinol Lett. 26(6):657-61.
Cubero J, Narciso D, Terrón P, Rial R, Esteban S, Rivero M, Parvez H, Rodríguez AB, Barriga C. 2007. Chrononutrition applied to formula milks to consolidate infants' sleep/wake cycle. Neuro Endocrinol Lett. 28(4):360-6.
Cubero J, Chanclón B, Sánchez S, Rivero M, Rodríguez AB, Barriga C. 2009. Improving the quality of infant sleep through the inclusion at supper of cereals enriched with tryptophan, adenosine-5'-phosphate, and uridine-5'-phosphate. Nutr Neurosci. 12(6):272-80.
Gila-Díaz A, Herranz Carrillo G, Cañas S, Saenz de Pipaón M, Martínez-Orgado JA, Rodríguez-Rodríguez P, López de Pablo ÁL, Martin-Cabrejas MA, Ramiro-Cortijo D, Arribas SM. 2020. Influence of Maternal Age and Gestational Age on Breast Milk Antioxidants During the First Month of Lactation. Nutrients. 12(9):2569.
Glynn LM, Davis EP, Schetter CD, Chicz-Demet A, Hobel CJ, Sandman CA. 2007. Postnatal maternal cortisol levels predict temperament in healthy breastfed infants. Early Hum Dev. ;83(10):675-81.
Grey KR, Davis EP, Sandman CA, Glynn LM. 2013. Human milk cortisol is associated with infant temperament. Psychoneuroendocrinology. 38(7):1178-85.
Hahn-Holbrook J, Saxbe D, Bixby C, Steele C, Glynn L. 2019. Human milk as "chrononutrition": implications for child health and development. Pediatr Res. 85(7):936-942.
Harada T, Hirotani M, Maeda M, Nomura H, and Takeuchi H. 2007. Correlation between breakfast tryptophan content and morning-evening in Japanese infants and students aged 0-15 yrs. J Physiol Anthropol. 26(2):201-7.
Hinde K, Skybiel AL, Foster AB, Del Rosso L, Mendoza SP, and Capitanio JB. 2015. Cortisol in mother’s milk across lactation reflects maternal life history and predicts infant temperament. Behavioral Ecology 26 (1): 269-281.
Katzer D, Pauli L, Mueller A, Reutter H, Reinsberg J, Fimmers R, Bartmann P, Bagci S. 2016. Melatonin Concentrations and Antioxidative Capacity of Human Breast Milk According to Gestational Age and the Time of Day. J Hum Lact. 32(4):NP105-NP110.
Pundir S, Wall CR, Mitchell CJ, Thorstensen EB, Lai CT, Geddes DT, Cameron-Smith D. 2017. Variation of Human Milk Glucocorticoids over 24 hour Period. J Mammary Gland Biol Neoplasia. 22(1):85-92.
Qin Y, Shi W, Zhuang J, Liu Y, Tang L, Bu J, Sun J, Bei F. 2019. Variations in melatonin levels in preterm and term human breast milk during the first month after delivery. Sci Rep. 2019 9(1):17984.
Sánchez CL, Cubero J, Sánchez J, Chanclón B, Rivero M, Rodríguez AB, and Barriga C. 2009. The possible role of human milk nucleotides as sleep inducers. Nutr Neurosci. 12(1):2-8.
van der Voorn B, de Waard M, van Goudoever JB, Rotteveel J, Heijboer AC, Finken MJ. 2016. Breast-Milk Cortisol and Cortisone Concentrations Follow the Diurnal Rhythm of Maternal Hypothalamus-Pituitary-Adrenal Axis Activity. J Nutr. 146(11):2174-2179.
Content of "Breastfeeding and baby formula" last modified 3/2021
Title image of infant feeding, mother concealed face by Nattakorn Maneerat / istock
image of mother bottlefeeding infant, sideview by PorporLing / shutterstock
image of father bottlefeeding infant by Halfpoint / shutterstock