Masshunter workstation software qualitative analysis version b 06

Serum metabolites of the Control and DKD groups at 6 and 15 weeks of age were extracted and analyzed by the Human Metabolome Technologies (HMT Inc., Yamagata, Japan) method. Briefly, methanol containing Internal Standard Solution 1 (HMT Inc.) was added to the sera and mixed well. Subsequently, 200 µL of Milli‐Q water (Merck Millipore, Burlington, MA, USA) and 500 µL of chloroform were added and centrifuged at 2300 g for 10 min at 4 °C. The upper aqueous layer was collected and then filtrated with a Millipore 5‐kDa cutoff filter and dried. The metabolites were resuspended in 25 µL of Milli‐Q water containing Internal Standard Solution 3 (HMT Inc) and applied to CE‐MS. All CE‐MS experiments were performed using an Agilent 7100 CE capillary electrophoresis system (Agilent Technologies, Santa Clara, Waldbronn, Germany) connected to an Agilent 6530 accurate Q‐TOF‐MS system (Agilent Technologies Inc., Santa Clara, CA, USA).
The data obtained by CE‐MS analysis were preprocessed using masshunter Workstation Software Qualitative Analysis version B.06.00 (Agilent Technologies Inc.). Each metabolite was identified and quantified based on the peak information including m/z, migration time, and peak area. The metabolite peaks were analyzed statistically using the mass profiler professional software (Agilent Technologies Inc.).