"The pVDTS workflow for solid whole larval samples. An accurately measured volume ( V f ) of saline containing 13 C-formate at a known concentration [ f ] 0 is transferred to a number of larvae ( n ) in a <t>microcentrifuge</t> tube (a). An accurately measured volume ( V t ) is then removed from the larvae in the microcentrifuge tube for the ""unopened"" sample (b). Larvae are then homogenized using a motorized pellet pestle and the microcentrifuge tube as a mortar (c)—this will be the ""opened"" sample. An accurately measured volume ( V t ) is then removed from the homogenized larvae and transferred to a known volume ( V D ) of chloroform/methanol/water (green) containing a fixed concentration [ D ] 0 of DSS (d). The remaining steps (e–i) are the same as (e–i) in Figure 1 . V WL : volume of homogenized whole larva. [X] WL : concentration of metabolite X in whole larval homogenate. I ′ 1 f and I ′ 2 f : ""unopened"" and ""opened"" sample signal intensities as per Figure 1 ."
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