Table 2.

Absolute quantification of HMI components in fresh and spent media after 56 h of trypanosome growth to 4 × 10⁶ cells ml^−1f

Component	Exptl concn (μM)a				Theoretical concn (μM)
	HMI		CMM		HMI11 reciped	CMM recipe	Human bloode	Human CSFe
	Fresh	Spent	Fresh	Spent
d-Glucosea					25,000	10,000	4,817	3,357
l-Cysteinea	3,349,188 ± 1,426,490	661,446 ± 147,585	3,987,853 ± 435,789	254,831 ± 85,829	1,504	1,000	99	0
l-Glutamine	4,002 ± 238	3,382 ± 191	727 ± 11	289 ± 6	3,998	1,000	550	490
l-Phenylalanine	327 ± 16	224 ± 8	76 ± 1	1.3 ± 0.1	400		79	11
l-Tryptophan	87 ± 5	73 ± 3	21 ± 1	0.4 ± 0.1	78		54	6
l-Tyrosine	1,013 ± 60	951 ± 44	194 ± 5	115 ± 6	574		89	11
l-Methionine	195 ± 8	188 ± 7	39 ± 1	22 ± 1	201		29	4
l-Arginine	692 ± 40	715 ± 28	225 ± 1	219 ± 9	482		110	21
l-Leucinea,b	48,270,601 ± 13,204,959	44,388,168 ± 4,445,117	17,478,123 ± 2,079,608	4,996,443 ± 584,426	801		158	14
l-Isoleucinea,b					801		69	7
l-Valine	440 ± 30	517 ± 69	385 ± 22	404 ± 44	803		234	19
l-Lysine	1,302 ± 65	1,382 ± 56	255 ± 4	231 ± 9	999		236	27
l-Threonine	899 ± 52	895 ± 42	102 ± 2	72 ± 4	798		155	31
l-Serine	553 ± 33	555 ± 27	193 ± 7	156 ± 10	400		145	30
l-Proline	483 ± 60	572 ± 57	128 ± 5	115 ± 5	348		190	3
l-Histidine	241 ± 12	225 ± 9	52 ± 1	46 ± 3	271		106	16
Glycine	482 ± 25	542 ± 25	179 ± 4	183 ± 9	400		266	8
l-Glutamate	729 ± 39	791 ± 32	179 ± 3	216 ± 10	510		76	13
l-Asparagine	276 ± 13	284 ± 10	91 ± 1	88 ± 4	189		50	7
l-Aspartate	338 ± 19	367 ± 15	100 ± 2	103 ± 6	226		21	2
l-Alanine	514 ± 41	1,263 ± 59	256 ± 6	793 ± 34	281		448	32
l-Cystinea	33,127 ± 57,378	<LODc	152,042 ± 14,351	<LODc	379		101	0.2
Pyruvate	937 ± 17	9,408 ± 503	4.7 ± 1.2	11,525 ± 2,053	1,295		60	98
Hypoxanthine	965 ± 47	983 ± 37	4.4 ± 0.1	3.8 ± 0.2	1,000		8	4
Thymidinea	154,748 ± 29,103	507,275 ± 74,464	<LODc	<LODc	161		0.2
myo-Inositola					40		24	138
Cholinea	7,999,054 ± 1,371,732	8,557,781 ± 1,051,370	3,691,885 ± 172,624	3,698,509 ± 306,766	38		9	6
Nicotinamide	4 ± 0.7	8.9 ± 1.0	0.02 ± 0.001	0.04 ± 0.01	33		0.4
Pyridoxala	420,434 ± 155,128	460,607 ± 178,818	<LODc	2,726 ± 506	24		0.3
Pantothenate	26 ± 2	29 ± 2	2.0 ± 0.1	1.8 ± 0.1	18		4	3
Thiaminea	3,643,586 ± 61,195	3,665,175 ± 218,103	3,531,013 ± 41,082	3,369,323 ± 26,130	15		0.1	0.07
Folic acida	150,002 ± 37,145	170,635 ± 65,126	<LODc	<LODc	9		0.03	0.02
Riboflavina	55,730 ± 9,975	54,715 ± 4,636	<LODc	<LODc	1		0.3	0.4
Biotina	10,264 ± 1,217	11,764 ± 1,234	9,819 ± 1,057	10,539 ± 735	0.053		0.002	0.0005
Cyanocobalamina					0.010		0.0003	0.00002

^aAbsolute quantification of these metabolites could not be performed by the LC-MS methods and internal standards that were used in this study. Where possible, normalized peak intensities (arbitrary units) are shown in italics to allow comparison of experimental relative abundances.

^bLeucine and isoleucine peaks overlapped. Peak intensity represents the sum of both metabolites.

^cA peak was not detected or was below the limit of detection (<LOD), which was set at 1,000 arbitrary units.

^dTheoretical concentrations of HMI11 components were taken from http://tryps.rockefeller.edu/trypsru2_culture_media_compositions.html.

^eTheoretical concentrations in human blood and cerebrospinal fluid (CSF) are average values for healthy adults obtained from www.hmdb.ca.

^fn = 5.