This study aims to evaluate whether eating biscuits made with mature okra flour can help young adults feel fuller (enhance satiety) and change certain blood markers compared to eating standard biscuits. Mature okra is a nutrient-dense agricultural by-product that is often discarded due to its tough texture. For this study, mature okra pods are dried and ground into a flour, which is then used to replace 10% of the wheat flour in a custom biscuit formulation. Participants will take part in a single-blind, randomised trial where they will consume either the okra-incorporated biscuits or the standard control biscuits on separate occasions. Researchers will measure participants' perceived fullness over 165 minutes using a visual rating scale. Blood samples will also be collected at specific intervals during this time to evaluate blood glucose levels, appetite-related hormones, and metabolic responses.
Okra is one of the top ten most consumed vegetable crops by Malaysians, with 1.7kg per capita consumption as reported by the Department of Statistic Malaysia in 2023. Among the total 65.6 thousand tonnes of output available, around 7.7% of the okra is turned into waste and partially fed to the livestock. Mature okra becomes unmarketable and turns into waste because it develops a tougher, more fibrous texture and a less desirable taste, unlike tender, immature okra, which is typically preferred for consumption. Studies in the literature found that mature okra also possesses comparable nutrient content or is even better than immature okra in certain aspects. Incorporation of agricultural waste, for instance, mature okra, into food products can offer significant health advantages besides reducing waste and also promotes a sustainable future, aligning with the objectives of Sustainable Development Goals (SDGs) goal No.12. Besides, transforming mature okra and other agricultural by-products into functional or value-added foods is an urge to address environmental and unsustainable use of natural resources. This study aims to evaluate the impact of mature okra flour-incorporated biscuits (MOFB) on perceived satiety and metabolomic responses. Mature okra flour will be prepared by dehydrating and grinding okra pods, then incorporated into biscuits at 10% substitution of wheat flour. This ratio of mature okra flour incorporation is pre-determined based on the previous phase study, as this ratio of incorporation results in a better nutritional profile and an acceptable sensorial score. Subsequently, thirty healthy young adults will participate in a single-blind, randomised trial comparing MOFB and control biscuits. Satiety will be assessed using the SLIM scale and blood glucose measurements over 165 minutes post-consumption. Blood samples will be collected for hormone and metabolomic analyses using ELISA, RIA, and Liquid Chromatography-Mass Spectrometry (LC-MS). The findings will help determine whether MOFB enhances satiety and alters metabolic profiles, supporting its potential as a functional food for appetite regulation.
Study Type
INTERVENTIONAL
Allocation
RANDOMIZED
Purpose
OTHER
Masking
SINGLE
Enrollment
30
Standard biscuit formulation manufactured where 10% of the wheat flour is substituted with mature okra flour, designed to enhance the nutritional profile while maintaining acceptable sensory scores.
Standard biscuit formulation manufactured using 100% wheat flour, designed to match the sensory profile and appearance of the experimental biscuit without the inclusion of mature okra flour.
Faculty of Medicine and Health Sciences, Universiti Putra Malaysia (UPM)
Serdang, Selangor, Malaysia
RECRUITINGSatiety Response
The satiety response after biscuit consumption will be determined by the Satiety Labeled Intensity Magnitude (SLIM) scale. The SLIM scale consists of a 100mm vertical line with labelled phrases to indicate satiety: "greatest imaginable fullness", "extremely full", "very full", "moderately full", "slightly full", "neither hungry nor full", "slightly hungry", "moderately hungry", "very hungry", and "extremely hungry", and "greatest imaginable hunger". The first satiety survey is filled before breakfast (time 0), and the remaining surveys at 15, 45, 75, 105, 135 and 165 min later.
Time frame: Baseline (0 minutes) and post-consumption at 15, 45, 75, 105, 135, and 165 minutes
Change in Plasma Insulin Concentration
Plasma concentration of insulin will be measured using an enzyme-linked immunosorbent assay (ELISA) kit. This metric evaluates metabolic responses to the biscuit interventions.
Time frame: Fasting state (0 minutes), 75 minutes, and 165 minutes post-consumption
Change in Active Glucagon-Like Peptide-1 (GLP-1) Concentration
Plasma concentration of active GLP-1 will be measured using an enzyme-linked immunosorbent assay (ELISA) kit. This metric evaluates anorexigenic hormone responses to the biscuit interventions.
Time frame: Fasting state (0 minutes), 75 minutes, and 165 minutes post-consumption
Change in Plasma Peptide YY (PYY3-36) Concentration
Plasma concentration of PYY3-36 will be measured using an enzyme-linked immunosorbent assay (ELISA) kit. This metric evaluates anorexigenic hormone responses to the biscuit interventions.
Time frame: Fasting state (0 minutes), 75 minutes, and 165 minutes post-consumption
Change in Plasma Cholecystokinin (CCK) Concentration
Plasma concentration of CCK will be determined using a radioimmunoassay (RIA) kit. This metric evaluates anorexigenic hormone responses to the biscuit interventions.
Time frame: Fasting state (0 minutes), 75 minutes, and 165 minutes post-consumption
Change in Active Ghrelin Concentration
Plasma concentration of active ghrelin will be measured using an enzyme-linked immunosorbent assay (ELISA) kit. This metric evaluates orexigenic hormone responses to the biscuit interventions.
Time frame: Fasting state (0 minutes), 75 minutes, and 165 minutes post-consumption
Changes in Metabolite Profiles
For metabolomic analysis, the blood samples will be taken in the fasting state (0 min) and after the last satiety survey (165 min). Metabolites will be quantified by Liquid Chromatography-Mass Spectrometry (LC-MS) analysis, and the serum samples will be prepared according to In Vitro Diagnostics Research (IVDr) standard operating procedures.
Time frame: Fasting state (0 minutes) and 165 minutes post-consumption
Blood Glucose Response
The capillary blood glucose will be measured using the finger-prick method during the fasting state (time 0), and at 15, 45, 75, 105, 135 and 165 min later.
Time frame: Baseline (0 minutes) and post-consumption at 15, 45, 75, 105, 135, and 165 minutes
Ad Libitum Energy Intake at a Subsequent Meal
Following the 165-minute satiety evaluation session, participants will be served an ad libitum meal consisting of fried rice and requested to consume as much as they want until comfortably full. The weight of the food consumed will be calculated by subtracting the remaining food weight from the initial served weight using a digital weighing scale (accurate to 1.0 g). Total energy intake will be calculated by multiplying the weight of the food consumed by the specific caloric content of the fried rice (expressed in kilocalories).
Time frame: At the end of the satiety evaluation session (165 minutes post-biscuit consumption)
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