Introduction:

This tool is used to annotate compounds from the LC-MS peak table by matching mz and RT with the public/custom library. The best top five hits will be shown in the results. If you want to use the custom library for annotation, a two-column Tab-delimited text file is required with the first column as compound name and the second column as precise MZ.

Input files:

1.      Raw peak table file in Tab-delimited txt format, can be obtained from tool "metaMS.runLC". First column is the compound identifier, the others are samples.

For example:

ID

mz

rt

wt15

wt16

wt18

wt19

LC1

589.25

41.76407

0

0

0

9347.745

LC2

332

41.81412

0

99789.56

0

0

LC3

246.1

41.99718

0

50669.26

0

21915.15

LC4

244.1

42.03141

0

35808.77

0

34583.09

LC5

317

42.03826

0

11905.25

0

62753.99

Parameter:

1.        mz cutoff: A hit that difference of mz must be <=mz_cutoff.

2.        RT window: The acceptable retention time difference.

3.        ppm: Parts per million. numeric the relative error for matching peaks that is a window of user specified error (or the default 10) in ppm for each fragment mass.

4.        (|M- M0|¡Âm) ¡Á106 (ppm). ¡®M¡¯ is the measured value of the ion mass; ¡®M0¡¯ is the theoretical value of the ion mass; ¡®m¡¯, an integer, is the mass of the ion.

5.        For example, the molecular ion measured value of a compound is 364.2504, the theoretical value is 364.2509, and the mass measurement accuracy is: |364.2504-364.2509|¡Â364¡Á106=1.4ppm

6.        adducts type: There are several possible adducts and the recommended type that most commonly occurs is ¡°M+H¡± or ¡°M-H¡±.

Output files:

1.      'identified_pkTable.txt', identified total peak table file in Tab-delimited txt format, containing duplicate row names possibly.

For example:

ID

wt15

wt16

wt18

wt19

[3-(7-hydroxy-4-oxo-4H-chromen-2-yl)phenyl]oxidanesulfonic acid

0

99789.56

0

0

2-{[hydroxy(2,3,4-trimethoxyphenyl)methylidene]amino}acetic acid

0

50669.26

0

21915.15

N-Phenylacetyl pyroglutamic acid

0

35808.77

0

34583.09

1D-Myo-inositol 1,4-bisphosphate

0

11905.25

0

62753.99

Phenytoin

44555.52

95913.45

0

13436.41

 

2.      'identified_uniq_pkTable.txt', identified uniq peak table file in Tab-delimited txt format. When row names are duplication, the row with the maximum intensity will be retained.

 

3.      'detailed_information.txt', detailed information about query and database relationship in library searching.

Columns are:

'Query'

query name in inputs.

'Query_mz'

MZ of the query

'Best_hit'

name of the best hit

'Best_hit_adducts_type'

adducts type of the best hit

'Best_hits_mz'

MZ of the best hit

'Best_hit_delta_mz'

delta MZ of the best hit

'Best_hit_rt'

rt of the best hit

'Best_hit_delta_rt'

delta rt of the best hit

'Other_hits'

other hits in searching

'ther_hits_adducts_type'

adducts type of the other hit

'Other_hits_mz'

Other hits MZ

'Other_hits_delta_mz'

delta MZ of the other hit

'Other_hits_rt'

Other hits rt

'Other_hits_delta_rt'

delta rt of the other hit

Note:

1.     There is no retention time field in the public library and only mass spectrum information is used for annotation. For a custom library, this tool supports the joint annotation by precise MZ and retention time. Users can provide an in-house two-column Tab-delimited text file with the first column as compound name and the second column as precise MZ.

2.     The tool will identify all possible matching compounds based on all adduct types selected, sort them according to the matching score, and output them all as 'detailed_information.txt'. Also, the compound with the smallest MZ and RT deviation will be outputted as the final identified compound in 'identified_pkTable.txt' file.