Table 1.

Sample metabolite sets (attomoles/CFU) extracted from P. aeruginosa cell pellets using different methods and detected by ¹H NMR. Metabolites were identified and quantified via Chenomx NMR Suite (v8.0); I (50% methanol); II (100% methanol); III (100% water); IV (2:1 methanol:chloroform); nd = not detected. All extraction methods were performed in triplicate. Amino acids are designated using their respective 3 letter abbreviations; GABA = 4-aminobutyrate; ADP = adenosine diphosphate; AMP = adenosine monophosphate; NAD⁺ = nicotinamide adenine dinucleotide; NADP⁺ = nicotinamide adenine dinucleotide phosphate; UMP = uridine monophosphate.

Compound	Planktonic				Biofilm
	IV	I	II	III	IV	I	II	III
GABA	nd	0.372±0.022	0.159±0.013	0.488±0.050	0.300±0.004	0.352±0.025	0.226±0.035	1.229±0.101
ADP	0.209±0.040	0.047±0.001	0.097±0.005	0.083±0.015	0.023±0.016	0.158±0.031	0.117±0.007	0.229±0.043
AMP	0.369±0.056	0.318±0.004	0.110±0.003	0.168±0.020	0.624±0.016	0.528±0.016	0.375±0.048	0.734±0.132
Acetate	0.016±0.003	0.591±0.002	0.195±0.034	1.12±0.09	0.085±0.012	5.532±0.090	6.409±2.072	31.16±0.05
Ala	0.601±0.030	1.54±0.36	0.484±0.083	3.42±0.92	0.204±0.016	2.423±0.313	0.307±0.007	3.716±0.632
Betaine	0.182±0.041	0.614±0.000	0.393±0.021	0.299±0.018	0.198±0.008	0.312±0.021	nd	nd
Choline	0.020±0.004	0.056±0.004	0.010±0.001	0.02±0.00	0.018±0.000	0.065±0.009	0.022±0.006	0.102±0.002
Formate	0.037±0.001	1.42±0.10	0.393±0.040	0.075±0.006	2.61±0.05	3.265±0.335	1.310±0.243	2.467±0.086
Glu	1.22±0.17	3.23±0.22	1.20±0.03	2.61±0.59	2.48±0.09	5.566±0.369	3.043±0.263	6.540±0.677
Gln	0.400±0.001	0.296±0.114	0.432±0.080	0.706±0.185	nd	nd	nd	nd
Gly	0.278±0.013	0.837±0.133	0.114±0.019	1.04±0.30	0.213±0.021	0.944±0.079	0.260±0.052	1.619±0.282
His	0.036±0.001	0.096±0.025	0.05±0.02	0.095±0.003	nd	0.142±0.040	nd	0.498±0.119
Ile	0.191±0.009	0.912±0.257	0.407±0.068	1.50±0.47	0.082±0.016	1.328±0.182	0.158±0.001	1.796±0.661
Leu	0.463±0.022	1.78±0.51	1.02±0.15	3.94±0.94	0.242±0.004	2.940±0.392	0.497±0.041	4.614±1.440
Mannose	0.131±0.032	0.400±0.000	0.183±0.011	0.077±0.012	0.178±0.062	0.107±0.021	nd	nd
Methanol	0.093±0.043	0.116±0.007	0.02±0.01	0.030±0.008	0.172±0.037	0.976±0.124	0.135±0.005	0.219±0.063
Met	0.065±0.009	0.309±0.082	0.189±0.030	0.808±0.206	0.108±0.004	0.529±0.074	0.085±0.009	0.842±0.184
NAD+	0.117±0.001	0.167±0.059	0.173±0.004	nd	0.554±0.016	0.598±0.053	0.584±0.027	0.449±0.072
NADP+	0.118±0.012	0.095±0.000	0.05±0.01	0.039±0.005	0.149±0.004	0.165±0.019	0.174±0.008	0.146±0.032
Trp	nd	0.133±0.036	nd	nd	nd	0.196±0.059	nd	0.425±0.202
Tyr	0.111±0.008	0.465±0.147	0.250±0.046	1.04±0.27	0.088±0.016	0.714±0.053	0.119±0.024	1.046±0.321
UMP	0.106±0.027	0.097±0.002	0.02±0.00	0.063±0.008	0.140±0.033	0.111 ±0.014	0.074±0.011	0.443±0.075
Val	0.304±0.024	1.49±0.44	0.704±0.137	3.00±0.78	0.073±0.020	1.978±0.258	0.217±0.003	2.607±0.897