Recipient Organization
PUMPKIN BABY INC.
5 CHARLES WAY
LAWRENCEVILLE,NJ 08648
Performing Department
(N/A)
Non Technical Summary
PumpKin Baby is developing affordable, in-field (i.e., in homes, at milk banks, in neonatal intensive care units (NICUs)) diagnostic tools to assess breastmilk nutrition and shelf-life using rapid and easy-to-interpret color-reactive detection strips. This technology will provide guidance on whether freeze-thawed breastmilk is nutritionally adequate or even rancid at the time of use, helping to minimize breastmilk waste and ensure infants receive proper nutrition.To facilitate continued breastfeeding, nursing mothers often rely on freezer storage of expressed breastmilk, and those with lactation difficulties or babies in NICUs seek donor breastmilk, which is also stored frozen. However, the coldest available storage unit in most households - the kitchen freezer - cools to -20 °C, a temperature that does not adequately preserve the complex emulsified structure of human breastmilk.Milk composition analyses have revealed that banked donor milk fails to provide the energy density and macronutrients needed by very low-birthweight premature babies, indicating a critical need for effective characterization tools to identify and address nutritional gaps through supplementation. These findings undermine CDC guidelines, which state breastmilk can be stored in the freezer for up to six months and may be acceptable up to 12 months.To provide just-in-time information on the quality of stored breastmilk,PumpKin's inexpensive, easy-to-use, diagnostic test kit will measure 1) propensity for fat breakdown (i.e., lipase levels), 2) fat breakdown/rancidity in stored human breastmilk (i.e., glycerol levels), and 3) vitamin C content (i.e., antioxidant capacity). To develop thetechnology, this project will involve collection and characterization of breastmilk samples, measuring the key quality indicators (i.e., lipase, glycerol/free fatty acids (FFAs), and vitamin C) over time from freshly expressed to after months of storage. We will use colorimetric tools, similar in concept to pH strips, to allow for rapid testing of breastmilk samples on these quality indicators. The outputs of our developed test kits will be compared to standard laboratory assays to validate their accuracy. Development of the proposed technology would providecaregivers with accurate and timely information about shelf-life, condition after storage, and nutritional value of breastmilk, thus delivering insights on the quality of the breastmilk at time of use, and whether or not supplementation may be needed to meet nutritional guidelines.
Animal Health Component
34%
Research Effort Categories
Basic
33%
Applied
34%
Developmental
33%
Goals / Objectives
Major goals of the project are to develop a diagnostic test kit capable of measuring the following in breastmilk:1) the propensity for fat breakdown (i.e., lipase levels), 2) fat breakdown/rancidity in stored breastmilk (i.e., glycerol levels), and 3) vitamin C content.To achieve these goals, this project will involve characterization of human breastmilk samples to define the relevant concentrations for lipase, glycerol, and vitamin C that determine breastmilk shelf-life, fat content, rancidity status, and essential vitamin C content, respectively. Regarding development and validation of the three test kits (i.e., for lipase, glycerol, and vitamin C), we aim for the kits to produce accurate readouts within 2 minutes over a range of concentrations of the three test components. Accuracy of the readouts will be validated against results from standardlaboratory-grade assays, targeting demonstrations of statistical similarity (p<0.05) in the measurements from our colorimetric test kits and the laboratory-grade assays. We also aim to demonstrate low standard error between predicted and actual values for breastmilk shelf-life (SE<8%), fat content (SE<5%), and vitamin C deficiency levels (SE<12%).
Project Methods
First, we will establish the upper and lower bounds of acceptable and typical amounts of lipase, glycerol, and vitamin C in human breastmilk and their connections to nutrient content and frozen shelf-life. This analysis will be accomplished through characterization of breastmilk provided from a wide range of donors and across the fresh- to freeze/thawed life cycle of days to months. We will use standard laboratory assays, including the Themo EnzChek Lipase Substrate assay for lipase; Promega Glycerol-Glo Assay and Caymen Chem FFA Fluorometric Assay Kit for glycerol/rancidity; and Sigma Ascorbic Acid Assay Kit for vitamin C.The characterization effort will enable us to understand the interpersonal differences in breastmilk lipase, glycerol, and vitamin C levels so we can define concentration ranges to drive storage and supplementation guidelines.Second, a suite of affordable, at-home colorimetric diagnostic test strips will be developed to measure the amounts of lipase, glycerol, and vitamin C content in breastmilk samples. Test kit development will build on preliminary work using colorimetric tools to assess enzymatic reactions characterizing the levels of lipase, glycerol, and vitamin C in breastmilk samples.Assay developmentto deliver rapid, accurate readings will be combined with learnings from the above characterization work to connect measurements to practical guidelines.Finally, the accuracy of the developed test kits will be validated using breastmilk samples assayed using both our approach and gold-standard laboratory-grade measurement tools. Collectively, this work will provide parents, milk banks and their individual and NICU recipients with easy-to-use, and inexpensive tools to assess the safety and nutritional value of their breastmilk.