# Effects of Dietary Ammonium Chloride Supplementation on the Lipidome and Volatile Flavor Compounds in the Subcutaneous Adipose Tissue of Tibetan Sheep

## TL;DR

This study examines how adding ammonium chloride to Tibetan sheep diets affects fat composition and meat flavor.

## Contribution

The study reveals how different levels of ammonium chloride supplementation alter lipid metabolism and flavor compounds in Tibetan sheep.

## Key findings

- High ammonium chloride levels increased saturated fats and phospholipids while reducing unsaturated fatty acids and beneficial flavor compounds.
- Ammonium chloride supplementation above 2.24% correlated with abnormal lipid accumulation and elevated ammonia in adipose tissue.
- Changes in lipid metabolism were linked to reduced synthesis of key aroma compounds like 2-propanone and 2-butanone.

## Abstract

As a source of non-protein nitrogen, ammonium chloride (NH4Cl) is widely utilized in ruminant diets to reduce feed costs. However, the impact of its supplementation level on the flavor of sheep meat remains unclear, despite the known influence of fat on meat flavor. This study aimed to investigate the effects of dietary NH4Cl supplementation levels on the lipidome and flavor compounds of subcutaneous adipose tissue in Tibetan sheep, providing a scientific basis for dietary optimization in Tibetan sheep farming. Eighty 2-month-old early-weaned Tibetan lambs were selected and randomly allocated into four groups, fed diets supplemented with 0% (N0 group), 1.49% (N1 group), 2.24% (N2 group), and 3.01% (N3 group) NH4Cl for an experimental period of 105 days. The study conducted histomorphological observations, lipidomics analysis, and determination of flavor compounds. The results showed that NH4Cl supplementation significantly reduced (p < 0.05) the contents of various unsaturated fatty acids and n-3 polyunsaturated fatty acids (n-3 PUFA) in the subcutaneous adipose tissue of Tibetan sheep. Specifically, the total saturated fatty acid (total SFA) content in the N3 group was significantly higher than that in the other groups, while the total monounsaturated fatty acid (total MUFA) content was significantly lower than that in the N1 and N2 groups (p < 0.05). The absolute contents of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and the sum of triglycerides (TGs) and diglycerides (DGs) in the N3 group were significantly higher (p < 0.05) than those in the other groups. Regarding flavor compounds, the contents of ketone aroma compounds, such as 2-propanone and 2-butanone monomer, were significantly higher (p < 0.05) in the N0 group than in the other groups. The ammonia content in the N1 and N3 groups was significantly higher (p < 0.05) than that in the N0 and N2 groups, while the allyl sulfide content in the N2 group was significantly higher (p < 0.05) than that in the other groups. Correlation analysis revealed that the majority of TG and DG differential lipids were significantly positively correlated with allyl sulfide, and most differential lipids belonging to the PC, PE, and hexosylceramide (Hex1Cer) classes were significantly positively correlated with ammonia (|r| ≥ 0.80, p < 0.01). Conversely, PC (16:0_18:3) exhibited significant negative correlations with multiple beneficial aroma compounds (|r| ≥ 0.80, p < 0.01). The study indicates that dietary NH4Cl supplementation levels exceeding 2.24% are associated with alterations in lipid metabolism and reduced synthesis of unsaturated fatty acids and beneficial flavor compounds, such as 2-propanone and 2-butanone, in subcutaneous adipose tissue. This is also associated with the abnormal accumulation of phospholipids and ceramides, which correlate strongly with elevated ammonia concentrations in adipose tissue and the generation of oxidation products such as propanal, potentially compromising meat flavor quality.