Table 2 MRM parameters and validation data for the quantitation of 101 SPLs in wild Cordyceps and its mycelia.

From: Quantitative profiling of sphingolipids in wild Cordyceps and its mycelia by using UHPLC-MS

Class

Subclass

No.

Name

RT (min)

MRM transitions (m/z)

Fragmentor (v)

CE* (v)

Precision and stability (RSD%)£

Injection precision

Intra-day precision

Inter-day precision

Analyte’s stability

System’s stability

Sphingoid bases

So

IS-1

So (d17:1)

6.37

286.3 → 268.3

80

5

1.17

1.25

2.02

2.04

2.88

1

So (d14:2)

3.76

242.2 → 224.2

80

5

ND$

ND

ND

ND

2.51

2

So (d14:1)

5.30

244.2 → 226.1

80

5

0.74

2.85

2.18

2.50

2.43

3

So (d16:1)

5.95

272.3 → 254.2

80

5

ND

ND

ND

ND

4.68

4

So (d17:2)

7.11

284.3 → 266.2

80

5

ND

ND

ND

ND

3.69

5

So (d18:2)

6.50

298.3 → 280.3

80

5

0.98

5.08

6.59

3.97

2.45

6

So (d18:1)

6.78

300.3 → 282.3

80

5

0.41

1.85

2.10

1.90

2.99

7

So (t18:1)

6.38

316.3 → 298.3

80

5

0.55

1.16

1.09

2.69

2.30

8

So (t18:1) isomer

6.90

316.3 → 298.3

80

5

0.52

1.19

1.43

4.40

2.35

9

So (d20:1)

9.41

328.3 → 310.3

80

5

0.35

1.80

1.86

2.09

6.36

10

So (t22:2)

9.13

370.3 → 352.3

80

5

1.15

12.19

14.89

1.57

3.37

S1P

IS-2

S1P (d17:1)

6.55

366.2 → 250.3

105

10

0.30

0.94

1.74

4.79

5.40

11

S1P (d18:1)

9.03

380.3 → 362.3

105

10

1.02

4.67

2.43

2.54

5.76

S1Po#

12

S1Po (d18:1)

6.55

467.4 → 449.3

105

10

ND

ND

ND

ND

5.83

Sa

IS-3

Sa (d17:0)

6.57

288.3 → 270.3

110

20

0.49

2.15

2.19

1.37

4.34

13

Sa (d16:0)

6.24

274.3 → 256.3

110

20

ND

ND

ND

ND

4.51

14

Sa (m18:0)

7.12

286.3 → 268.3

110

20

ND

ND

ND

ND

3.72

15

Sa (d17:0) isomer

5.51

288.3 → 270.3

110

20

ND

ND

ND

ND

3.31

16

Sa (d18:0)

6.91

302.3 → 284.3

110

20

0.63

2.58

3.41

4.02

3.88

17

Sa (t18:0)

6.61

318.3 → 300.3

110

20

1.03

0.57

0.61

0.83

4.48

18

Sa (t18:0) isomer

6.32

318.3 → 300.3

110

20

0.58

2.43

2.06

6.09

3.11

Ceramides

Cer

IS-4

Cer (d18:1/12:0)

10.96

482.5 → 264.3

130

25

1.88

3.92

4.03

3.57

5.41

19

Cer (d14:1/22:0)

14.64

566.5 → 208.2

130

25

1.32

3.48

3.47

11.06

12.46

20

Cer (d18:1/2:0)

9.04

342.3 → 264.3

130

25

5.60

12.69

11.98

4.50

4.86

21

Cer (d18:1/14:2)

11.06

506.5 → 264.3

130

25

0.53

3.79

3.89

4.72

10.21

22

Cer (d18:1/14:1)

11.15

508.5 → 264.3

130

25

2.99

7.11

9.99

16.17

5.95

23

Cer (d18:1/14:0)

11.85

510.5 → 264.3

130

25

2.17

4.63

4.19

18.05

10.86

24

Cer (d18:1/15:0)

12.33

524.5 → 264.3

130

25

0.50

1.55

1.36

9.58

14.48

25

Cer (d18:1/16:2)

11.84

534.5 → 264.3

130

25

2.43

3.31

3.83

12.62

10.23

26

Cer (d18:1/16:1)

12.43

536.5 → 264.3

130

25

0.54

1.26

3.14

4.72

9.47

27

Cer (d18:1/16:0)

12.92

538.5 → 264.3

130

25

1.50

2.80

6.05

9.14

11.30

28

Cer (d18:1/17:1)

13.03

550.5 → 264.3

130

25

1.78

4.85

4.33

15.91

13.69

29

Cer (d18:1/18:2)

13.45

562.5 → 264.3

130

25

1.12

5.98

6.29

7.52

14.02

30

Cer (d18:1/18:1)

13.57

564.5 → 264.3

130

25

0.78

0.95

1.20

4.06

9.26

31

Cer (d18:1/18:0)

14.36

566.5 → 264.3

130

25

1.72

3.04

4.54

8.21

12.44

32

Cer (d18:1/20:1)

15.86

592.6 → 264.3

130

25

1.01

8.45

7.15

6.96

20.58

33

Cer (d18:1/22:0)

18.20

622.6 → 264.3

130

25

4.88

10.12

8.26

17.27

19.05

34

Cer (d18:1/23:0)

19.10

636.6 → 264.3

130

25

2.82

4.01

7.49

5.38

14.47

35

Cer (d18:1/24:0)

20.10

650.6 → 264.3

130

25

4.22

4.78

4.05

3.73

13.24

36

Cer (d18:1/25:0)

19.25

664.7 → 264.3

130

25

ND

ND

ND

ND

14.40

37

Cer (d18:1/26:1)

19.23

676.7 → 264.3

130

25

5.08

2.38

2.94

7.04

14.17

38

Cer (d18:2/15:0)

11.83

522.5 → 262.3

130

25

2.53

4.35

6.87

14.73

11.54

39

Cer (d18:2/16:1)

11.85

534.5 → 262.3

130

25

0.73

6.26

3.63

7.28

2.68

40

Cer (d18:2/16:0)

12.38

536.5 → 262.3

130

25

0.39

1.49

1.17

3.74

11.23

41

Cer (d18:2/18:1)

13.50

562.5 → 262.3

130

25

ND

ND

ND

ND

10.07

42

Cer (d18:2/23:0)

18.18

634.6 → 262.3

130

25

ND

ND

ND

ND

21.35

43

Cer (d18:2/24:0)

19.06

648.6 → 262.3

130

25

4.01

4.34

5.62

5.10

13.95

44

Cer (d19:2/16:0)

12.76

550.5 → 276.3

130

25

1.04

1.50

2.49

5.83

10.33

45

Cer (d18:1/16:1(OH))

12.32

552.5 → 264.3

130

25

3.06

4.12

4.46

9.65

14.78

46

Cer (d18:1/16:0(OH))

12.40

554.5 → 264.3

130

25

1.34

0.73

0.71

6.06

11.96

47

Cer (d18:1/17:1(OH))

15.36

566.5 → 264.3

130

25

2.36

2.68

3.95

6.35

14.16

48

Cer (d18:1/18:1(OH))

14.53

580.5 → 264.3

130

25

0.31

2.22

3.70

4.98

14.72

49

Cer (d18:1/18:0(OH))

13.55

582.5 → 264.3

130

25

ND

ND

ND

ND

8.01

50

Cer (d18:1/19:1(OH))

12.55

594.5 → 264.3

130

25

1.66

2.36

2.18

7.49

12.06

51

Cer (d18:1/22:0(OH))

17.11

638.6 → 264.3

130

25

2.40

6.06

10.11

19.27

19.71

52

Cer (d18:2/16:0(OH))

11.84

552.5 → 262.3

130

25

2.88

5.74

14.77

19.71

13.98

53

Cer (d18:2/18:1(OH))

11.52

578.5 → 262.3

130

25

ND

ND

ND

ND

12.92

54

Cer (d19:2/16:0(OH))

12.20

566.5 → 276.3

130

25

0.35

3.57

5.49

2.41

6.30

55

Cer (d18:0/15:0)

12.75

526.5 → 266.3

130

25

1.03

7.52

7.17

15.16

12.59

56

Cer (d18:0/16:1)

13.23

538.5 → 266.3

130

25

2.35

5.60

4.32

10.49

9.24

57

Cer (d18:0/16:0)

13.35

540.5 → 266.3

130

25

1.77

5.60

4.49

11.40

9.75

58

Cer (d18:0/18:0)

14.63

568.6 → 266.3

130

25

ND

ND

ND

ND

14.79

59

Cer (d18:0/26:0)

20.82

680.7 → 266.3

130

25

ND

ND

ND

ND

14.20

60

Cer (t18:1/18:0)

12.99

582.5 → 262.3

130

25

ND

ND

ND

ND

14.17

61

Cer (t18:1/22:1)

16.15

636.6 → 262.3

130

25

ND

ND

ND

ND

14.48

62

Cer (t18:1/22:0)

15.64

638.6 → 262.3

130

25

5.95

22.80

21.75

17.75

18.08

63

Cer (t18:0/16:0)

12.28

556.5 → 264.3

130

25

1.02

3.71

4.67

9.28

10.46

64

Cer (t18:0/22:1)

16.12

638.6 → 264.3

130

25

ND

ND

ND

ND

21.05

65

Cer (t18:0/22:0)

16.13

640.6 → 264.3

130

25

0.43

9.40

8.70

7.21

14.36

66

Cer (t18:0/24:0)

17.45

668.7 → 264.3

130

25

1.76

2.15

2.66

4.83

21.36

67

Cer (t18:0/18:0(OH))

12.98

600.6 → 264.3

130

25

2.68

4.36

4.53

11.48

12.06

68

Cer (t18:0/22:0(OH))

15.48

656.6 → 264.3

130

25

1.17

3.70

2.89

5.04

14.51

69

Cer (t18:0/24:0(OH))

16.83

684.7 → 264.3

130

25

2.32

2.59

2.43

18.20

22.32

Glycosphingolipids

HexCer

IS-5

GlcCer (d18:1/12:0)

10.40

644.5 → 264.3

130

30

1.99

1.09

1.04

4.79

5.89

70

HexCer (d18:1/16:0)

12.21

700.6 → 264.3

130

30

1.53

6.73

9.43

7.96

14.79

71

HexCer (d18:2/16:0)

11.52

698.6 → 262.3

130

30

1.20

2.91

2.01

4.81

4.81

72

HexCer (d18:2/16:0(OH))

11.25

714.6 → 262.3

130

30

2.15

6.51

7.11

5.81

14.71

73

HexCer (d19:2/15:0)

11.46

698.6 → 276.3

130

30

1.93

6.63

5.60

5.33

6.50

74

HexCer (d19:2/16:0(OH))

11.75

728.6 → 276.3

130

30

0.50

11.97

15.09

4.32

8.65

75

HexCer (d19:2/17:0(OH))

12.00

742.6 → 276.3

130

30

5.44

2.33

2.42

23.88

20.74

76

HexCer (d19:2/18:0(OH))

12.53

756.6 → 276.3

130

30

0.58

3.80

3.08

12.18

10.77

77

HexCer (t19:1/16:1)

11.20

728.6 → 276.3

130

30

ND

ND

ND

ND

10.09

Phosphosphingolipids

SM

IS-6

SM (d18:1/12:0)

10.37

647.5 → 184.1

170

20

1.87

2.02

1.72

5.38

5.40

78

SM (d18:1/14:0)

11.1

675.5 → 184.1

170

20

0.39

2.83

3.51

2.44

3.63

79

SM (d18:1/16:0)

12.00

703.6 → 184.1

170

20

0.52

1.33

2.18

2.3

4.9

80

SM (d18:1/18:0)

13.25

731.6 → 184.1

170

20

0.83

1.94

2.06

3.6

4.99

81

SM (d18:1/20:0)

14.51

759.6 → 184.1

170

20

1.06

5.48

4.76

5.08

10.22

82

SM (d18:1/22:0)

15.61

787.7 → 184.1

170

20

5.52

4.99

4.81

2.1

11.33

83

SM (d18:1/24:0)

16.91

815.7 → 184.1

170

20

2.12

3.65

2.42

2.09

14.39

84

SM (d18:1/16:1)

11.66

701.6 → 184.1

170

20

1.07

2.15

2.05

3.98

3.62

85

SM (d18:1/20:1)

13.90

757.6 → 184.1

170

20

1.75

2.8

3.48

3.14

9.43

86

SM (d18:1/22:1)

15.12

785.7 → 184.1

170

20

0.47

4.6

6.32

4.76

7.39

87

SM (d18:2/18:0)

12.75

729.6 → 184.1

170

20

1.4

6.59

6.9

7.41

5.37

88

SM (d18:2/18:1)

11.81

727.6 → 184.1

170

20

ND

ND

ND

ND

7.42

89

SM (d18:1/18:1(OH))

12.38

745.6 → 184.1

170

20

1.35

2.82

3.91

5.21

7.03

90

SM (d18:2/16:0(OH))

11.07

717.6 → 184.1

170

20

1.43

1.33

0.86

4.21

4.13

91

SM (d33:1)

11.56

689.6 → 184.1

170

20

2.79

5.38

4.6

2.57

7.44

92

SM (d39:2)

14.56

771.6 → 184.1

170

20

2.68

5.07

4.16

4.51

11.93

93

SM (d42:2)

16.14

813.7 → 184.1

170

20

1.65

2.82

2.77

1.49

10.5

94

SM (d34:0)

12.5

705.6 → 184.1

170

20

1.29

5.33

3.96

6.85

6.14

95

SM (d36:0)

13.84

733.6 → 184.1

170

20

1.91

3.23

3.04

5.74

7.85

96

SM (d38:0)

14.90

761.7 → 184.1

170

20

ND

ND

ND

ND

11.75

PI-Cer#

97

PI-Cer (d18:0/16:0(OH))

10.90

798.5 → 538.5

170

30

ND

ND

ND

ND

5.50

98

PI-Cer (t18:0/22:0(OH))

12.84

898.6 → 638.6

170

30

ND

ND

ND

ND

14.83

99

PI-Cer (t18:0/24:0(OH))

13.95

926.7 → 666.6

170

30

1.73

5.26

4.66

12.65

13.32

MIPC#

100

MIPC (t18:0/22:0(OH))

12.60

1060.7 → 638.6

170

40

ND

ND

ND

ND

20.32

101

MIPC (t18:0/24:0(OH))

13.68

1088.7 → 666.6

170

40

2.37

19.15

23.55

20.13

11.82

  1. *CE, Collision Energy. £Precisions and stabilities of endogenous SPLs in validation samples (Hirsutella sinensis) were calculated according to the quantitative data, while precisions and stabilities of 6 IS were calculated using peak area data; system’s stability was determined using QC sample. $ ND, Not detected in validation samples. # The IS for quantitation of S1Po was IS-2; the IS for quantitation of PI-Cer and MIPC was IS-4.
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