key: cord-1023093-5bystg08
authors: Sharma, Nupur; Bhat, Sadam H.; Tripathi, Gaurav; Yadav, Manisha; Mathew, Babu; Bindal, Vasundhra; Sharma, Shvetank; Gupta, Ekta; Singh, Jaswinder; Sarin, Shiv Kumar
title: Global metabolome profiling of COVID-19 respiratory specimen using High Resolution Mass Spectrometry (HRMS)
date: 2021-12-03
journal: STAR Protoc
DOI: 10.1016/j.xpro.2021.101051
sha: bdd1929bd76d8e3de8bff70c09cde7f04d9ce612
doc_id: 1023093
cord_uid: 5bystg08

Here we describe a protocol for identifying metabolites in respiratory specimens of patients that are SARS-CoV-2 positive, SARS-CoV-2 negative, or H1N1 positive. This protocol provides step-by-step instructions on sample collection from patients, followed by metabolite extraction. We use ultra high-pressure liquid chromatography (UHPLC) coupled High Resolution Mass Spectrometry (HRMS) for data acquisition and describe the steps for data analysis. The protocol was standardized with specific customization for SARS-CoV-2 containing respiratory specimens

Samples are analyzed in single runs for positive and negative modes using the 8 parameters shown in Table 1 .

Hypersil GOLD™ C18 Selectivity HPLC Column is used as stationary phase to 11 separate respiratory metabolites. Column Specifications are mentioned in Table 2 .

The mobile phases consist of (A) Acetonitrile with 1% formic acid and (B) Milli-Q-14 water with 0.1% formic acid, used for gradient elution (Table 3) . Step-by-step method details 2 An outline of the procedures is described in Figure 1 . A total of 500μL respiratory specimens 3 was heat-inactivated using heat block at 95°C for 15min in a BSL3 lab. Heat inactivated 4 respiratory specimen was subjected to homogenization by probe sonicator (steps 1-6). it is essential to homogenize the samples. This step provides you MS.raw file for each sample for further analysis. MS.raw file is mapped on different metabolome databases for metabolite annotation that is 10 exported in excel sheet for statistical analysis in order to interpret the data. The study did not generate any materials.

Data and code availability 8 The processed data is provided as supplemental information, and the raw data for the 9 manuscript is available on request to the lead contact Dr. Jaswinder Singh Maras 10 (jassi2param@gmail.com). 20 There is no conflict of interest from any of the authors included in the manuscript. 

Heat inactivation of the severe acute CONTROL, C. F. D. & PREVENTION 2020. Interim guidelines for collecting, handling, and testing clinical 16 specimens for COVID-19

Temporal change in the plasma 18 metabolome profile is indicative of outcome in severe alcoholic hepatitis

Baseline urine metabolic phenotype in patients with severe alcoholic hepatitis and its 21 association with outcome