In search of an accurate and effective method to determine fatty acid composition in silkworm pupae oils, five methylation methods were evaluated for use in the gas chromatographic (GC) quantitation of fatty acid methyl esters (FAMEs), including one-step esterification catalyzed by an acidic (H2SO4 and BF3) or alkali catalyst (KOH and NaOCH3) and a two-step procedure catalyzed successively by KOH and H2SO4. These methods were comparatively adopted to quantify FAMEs in silkworm pupae oil using GC-MS and GC and then validate their precision, stability and average recovery rates. The results indicated that compared with the other four methyl esterification methods, two-step methylation effectively improves the synthesis yield of FAMEs, conserves agents, and eliminates the usage of potential harmful reagents. The proposed GC method has been validated, shows good accuracy and precision, and has been applied successfully to the quantification of FAMEs in several varieties of silkworm pupae oils. The short analytical run time leads to low costs and a fast chromatographic procedure. In summary, two-step pretreatment had superior performance, providing technical references for the determination and analysis of fatty acids in other oils.

Fatty acid synthase (FAS) is a key enzyme in the lipid synthesis pathway, however, its roles in insects remain largely unknown. Here, we firstly identified two FAS genes from the transcriptome dataset of the general cutworm Spodoptera litura, which is a destructive insect pest of many crops. Both SlFAS1 and SlFAS2 were highly expressed in third instar larvae and in their fat bodies. Then, we successfully silenced SlFAS1 in third instar larvae and the content of α-linolenic acid and triglyceride was significantly decreased. Besides that, the effect of FAS on the metamorphic development in S. litura was evaluated. The results indicate that after silencing SlFAS1, the survival rates of S. litura larvae decreased significantly compared to the control groups. Silencing SlFAS1 in fifth instar larvae resulted in more malformed pupae and adults, and the emergence rates were significantly reduced. Furthermore, the ecdysone content in the haemolymph of fifth instar larvae was significantly decreased after silencing SlFAS1. In addition, knocking down SlFAS1 significantly alters the expression of other key genes in the lipogenesis pathway, implying that FAS has an impact on the lipogenesis pathway. The present study deepens the understanding of FAS in insects and provides novel potential targets for managing insect pests.