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Immunosuppressants (EDITED) July 12.
2012-07-12T11:31:45-04:00
2012-07-12T11:31:45-04:00
2012-07-12T11:31:44-04:00
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(32 Nixon Road, Bolton, Ontario, Canada)Tj
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)]TJ
-10.779 -1.2 Td
[(spectrometer \(Bolton, ON Canada\) with a Shimadzu UFLC system. 20 \265L\
)35( of supernatant were loaded on a )]TJ
-0.001 -1.2 Td
(porous R1/20 pretreatment column \(30x2.1mm\) for on-line washing with w\
ater for 0.25 minutes at a liquid )Tj
0.01 -1.2 Td
[(flow rate of 3 mL/min, then eluted by an Imtakt Cadenza CD-C18HT)16( analytical column \(50x2.0mm, 3\265m\) at )]TJ
0 -1.2 TD
[(flow rate of 0.6 mL/min using Solvent )54(A)55( \(water:methanol = 98:2, v/v)74(, with 0.1% formic acid and 10mM am)]TJ
46.197 0 Td
(-)Tj
-46.188 -1.2 Td
[(monium acetate\) and Solvent B \(water:methanol = 2:98, v/v)73(, with 0.1% formic acid and 10mM ammonium )]TJ
-0.017 -1.2 Td
(acetate\))Tj
/TT1 1 Tf
-0.008 -2.4 Td
(Mass Spectrometry Conditions:)Tj
/TT0 1 Tf
[( Electrospray V)55(oltage: 5000V)91(, HSID: 150\260C, Nebulizer Gas: 450, Drying )]TJ
0.025 -1.2 Td
[(Gas: 120, Heating Gas: 350, Source )18(T)111(emperature: 325\260C. )]TJ
-0.068 -2.4 Td
(The total LC cycle time for an injection is 3 min and the sample analysi\
s time is 1.5 min with alternating )Tj
0.058 -1.2 Td
[(injections from each LC. )54(All the solvents used in this method are HPLC grade.)]TJ
/TT1 1 Tf
36 0 0 36 3529.3052 1297.1274 Tm
(Results: )Tj
/TT0 1 Tf
(This method covers a concentration range of three orders of magnitude fr\
om 0.2 )Tj
0.012 -1.2 Td
[(to 200 ng/mL)37( for )18(T)111(acrolimus, Sirolimus, Everolimus and 2 to 2000 ng/mL)35( for Cyclosporin )55(A, )]TJ
-0.013 -1.2 Td
(while maintaining good linearity \(R)Tj
20.988 0 0 20.988 4077.5479 1222.7153 Tm
(2)Tj
36 0 0 36 4089.2207 1210.7271 Tm
[( = 0.999\) with 1/x weighting. )17(The intraday and interday )]TJ
-15.537 -1.2 Td
[(variability for three levels QCs were all <7% and <1)72(1%, respectively)73(. No interference or )]TJ
-0.016 -1.2 Td
(cross contamination was observed.)Tj
0.971 0.736 0.017 0 k
/TT1 1 Tf
54 0 0 54 3993.9399 2154.772 Tm
(Calibration Curves)Tj
ET
q
0 2592 5184 -2592 re
W n
0 0 0 0.9 k
q 1 0 0 1 4876.127 2134.7041 cm
0 0 m
-1252.799 0 l
h
f
Q
q 1 0 0 1 4876.127 2138.2041 cm
0 0 m
-60.8 0 l
-219.489 0 l
-440.504 0 l
-688.281 0 l
-927.257 0 l
-1121.866 0 l
-1236.545 0 l
-1252.799 0 l
-1257.313 0 -1257.313 -7 -1252.799 -7 c
-1191.999 -7 l
-1033.31 -7 l
-812.295 -7 l
-564.518 -7 l
-325.541 -7 l
-130.933 -7 l
-16.254 -7 l
0 -7 l
4.514 -7 4.514 0 0 0 c
-60.8 0 l
-219.489 0 l
-440.504 0 l
-688.281 0 l
-927.257 0 l
-1121.866 0 l
-1236.545 0 l
-1252.799 0 l
-1257.313 0 -1257.313 -7 -1252.799 -7 c
-1191.999 -7 l
-1033.31 -7 l
-812.295 -7 l
-564.518 -7 l
-325.541 -7 l
-130.933 -7 l
-16.254 -7 l
0 -7 l
4.514 -7 4.514 0 0 0 c
f
Q
Q
BT
0 0 0 1 k
/TT0 1 Tf
36 0 0 36 3523.1011 588.7197 Tm
[(A)55( sensitive, reliable and accurate LC-MS/MS method was developed and vali\
dated for )]TJ
-0.006 -1.2 Td
[(quantification of )17(T)111(acrolimus, Sirolimus, Everolimus and Cyclosporin )52(A)55( in whole blood. )17(The )]TJ
-0.003 -1.2 Td
(use of an ESI/ESI novel dual source allows a sample analysis time of onl\
y 1.5 minutes, )Tj
0 -1.2 Td
[(which doubles the throughput. )17(This LC-MS/MS method requires simple sample )]TJ
0.001 -1.2 Td
(preparation and is well-suited for routine therapeutic drug monitoring o\
f )Tj
0.01 -1.2 Td
(immunosuppressive drugs.)Tj
26 0 0 26 3523.0928 323.5195 Tm
(Reference: [1] Christoph Seger)Tj
15.158 0 0 15.158 3881.5068 332.1777 Tm
(1)Tj
26 0 0 26 3889.937 323.5195 Tm
[(, et al. )56(A)55( rapid HPLC-MS/MS method for the simultaneous quantification of cyclospo\
rine )52(A, )]TJ
-14.101 -1.2 Td
[(tacrolimus, sirolimus and everolimus in human blood samples. Nature Prot\
ocols, V)51(ol 4, 526-534 \(2009\) )]TJ
-0.002 -1.2 Td
[([2] V)56(ogeser M, Sp\366hrer U. Pitfall in the high-throughput quantification of\
whole blood cyclosporin )51(A)55( using liquid chromatogra)]TJ
54.477 0 Td
(-)Tj
-54.467 -1.2 Td
[(phy-tandem mass spectrometry)73(. Clin Chem Lab Med. 43\(4\):400-402 \(2005\).)]TJ
-0.009 -1.2 Td
[([3] Sha Joshua )18(Y)92(e, et al, Method Development )17(T)37(ime Reduction In Clinical )53(Applications By Multiplexing )16(T)55(wo HPLC )55(Analyses )]TJ
0.001 -1.2 Td
[(Using a Dual Coaxial Flow Ion Source , 201)72(1 )55(ASMS meeting poster)]TJ
0 0 0 0 k
/TT1 1 Tf
58.3 0 0 58.3 966.3955 2402.1011 Tm
[(High Throughput Simultaneous )37(Analysis of Immunosuppressants by ESI/ESI Dual Source Coaxial Flow Ion S\
ource LC-MS/MS System)]TJ
ET
q
0 2592 5184 -2592 re
W n
266.061 764.075 130.063 -74.399 re
f*
396.123 764.075 265.938 -74.399 re
f*
662.061 764.075 150 -74.399 re
f*
812.061 764.075 144 -74.399 re
f*
0 0 0 0.8 k
266.061 689.676 130.063 -74.4 re
f*
396.123 689.676 122.907 -74.4 re
f*
519.03 689.676 143.031 -74.4 re
f*
662.061 689.676 150 -74.4 re
f*
812.061 689.676 144 -74.4 re
f*
0 0 0 0 k
266.061 615.275 130.063 -40.8 re
f*
396.123 615.275 122.907 -40.8 re
f*
519.03 615.275 143.031 -40.8 re
f*
662.061 615.275 150 -40.8 re
f*
812.061 615.275 144 -40.8 re
f*
0 0 0 0.8 k
266.061 574.476 130.063 -40.8 re
f*
396.123 574.476 122.907 -40.8 re
f*
519.03 574.476 143.031 -40.8 re
f*
662.061 574.476 150 -40.8 re
f*
812.061 574.476 144 -40.8 re
f*
0 0 0 0 k
266.061 533.676 130.063 -40.801 re
f*
396.123 533.676 122.907 -40.801 re
f*
519.03 533.676 143.031 -40.801 re
f*
662.061 533.676 150 -40.801 re
f*
812.061 533.676 144 -40.801 re
f*
0 0 0 0.8 k
266.061 492.875 130.063 -40.8 re
f*
396.123 492.875 122.907 -40.8 re
f*
519.03 492.875 143.031 -40.8 re
f*
662.061 492.875 150 -40.8 re
f*
812.061 492.875 144 -40.8 re
f*
0 0 0 0 k
266.061 452.075 130.063 -40.8 re
f*
396.123 452.075 122.907 -40.8 re
f*
519.03 452.075 143.031 -40.8 re
f*
662.061 452.075 150 -40.8 re
f*
812.061 452.075 144 -40.8 re
f*
0 0 0 0.8 k
266.061 411.275 130.063 -40.8 re
f*
396.123 411.275 122.907 -40.8 re
f*
519.03 411.275 143.031 -40.8 re
f*
662.061 411.275 150 -40.8 re
f*
812.061 411.275 144 -40.8 re
f*
0 0 0 0 k
266.061 370.476 130.063 -40.8 re
f*
396.123 370.476 122.907 -40.8 re
f*
519.03 370.476 143.031 -40.8 re
f*
662.061 370.476 150 -40.8 re
f*
812.061 370.476 144 -40.8 re
f*
0.746 0.676 0.668 0.898 K
2 w 1 j
q 1 0 0 1 266.0605 689.6758 cm
0 0 m
690 0 l
S
Q
Q
BT
0.746 0.676 0.668 0.898 k
/TT2 1 Tf
27.96 0 0 27.96 439.9502 716.5625 Tm
(L)Tj
0.666 0 Td
(o)Tj
0.502 0 Td
(a)Tj
0.502 0 Td
(d)Tj
0.557 0 Td
(i)Tj
0.279 0 Td
(n)Tj
0.557 0 Td
(g)Tj
0.502 0 Td
( )Tj
0.249 0 Td
(P)Tj
0.61 0 Td
(u)Tj
0.557 0 Td
(m)Tj
(p)Tj
4.055 0.605 Td
(E)Tj
0.666 0 Td
(l)Tj
0.279 0 Td
(u)Tj
0.557 0 Td
(t)Tj
0.334 0 Td
(i)Tj
0.279 0 Td
(n)Tj
0.557 0 Td
(g)Tj
0.502 0 Td
( )Tj
-2.865 -1.206 Td
(P)Tj
0.61 0 Td
(u)Tj
0.557 0 Td
(m)Tj
(p)Tj
0 0 0 0 k
-15.439 -1.455 Td
(T)Tj
0.632 0 Td
(i)Tj
0.279 0 Td
(m)Tj
0.828 0 Td
(e)Tj
0.443 0 Td
( )Tj
-2.259 -1.205 Td
(\()Tj
0.334 0 Td
(m)Tj
0.828 0 Td
(i)Tj
0.279 0 Td
(n)Tj
0.558 0 Td
(\))Tj
1.603 1.205 Td
(S)Tj
0.557 0 Td
(o)Tj
0.502 0 Td
(l)Tj
0.279 0 Td
(v)Tj
0.502 0 Td
(e)Tj
0.443 0 Td
(n)Tj
0.558 0 Td
(t)Tj
0.334 0 Td
( )Tj
0.187 -0.025 Td
(A)Tj
0.666 0 Td
( )Tj
-2.68 -1.18 Td
(\()Tj
0.334 0 Td
(%)Tj
(\))Tj
3.12 1.205 Td
(S)Tj
0.557 0 Td
(o)Tj
0.502 0 Td
(l)Tj
0.279 0 Td
(v)Tj
0.502 0 Td
(e)Tj
0.443 0 Td
(n)Tj
0.558 0 Td
(t)Tj
0.334 0 Td
( )Tj
0.245 0 Td
(B)Tj
0.666 0 Td
( )Tj
-2.85 -1.205 Td
(\()Tj
0.334 0 Td
(%)Tj
(\))Tj
3.668 1.205 Td
(S)Tj
0.557 0 Td
(o)Tj
0.502 0 Td
(l)Tj
0.279 0 Td
(v)Tj
0.502 0 Td
(e)Tj
0.443 0 Td
(n)Tj
0.558 0 Td
(t)Tj
0.334 0 Td
( )Tj
0.245 0 Td
(B)Tj
0.666 0 Td
( )Tj
-2.85 -1.205 Td
(\()Tj
0.334 0 Td
(%)Tj
(\))Tj
4.538 1.205 Td
(V)Tj
0.61 0 Td
(a)Tj
0.499 0 Td
(l)Tj
0.279 0 Td
(v)Tj
0.502 0 Td
(e)Tj
0.443 0 Td
( )Tj
-2.839 -1.205 Td
(P)Tj
0.612 0 Td
(o)Tj
0.502 0 Td
(s)Tj
0.39 0 Td
(i)Tj
0.279 0 Td
(t)Tj
0.334 0 Td
(i)Tj
0.279 0 Td
(o)Tj
0.502 0 Td
(n)Tj
0.746 0.676 0.668 0.898 k
/TT3 1 Tf
-21.908 -1.459 Td
(0)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(0)Tj
0.502 0 Td
(1)Tj
3.395 0 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
4.254 0 Td
(0)Tj
4.74 0 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
3.267 0 Td
(W)Tj
0.863 0 Td
(a)Tj
0.443 0 Td
(s)Tj
0.39 0 Td
(h)Tj
0.502 0 Td
(i)Tj
0.279 0 Td
(n)Tj
0.502 0 Td
(g)Tj
0 0 0 0 k
-21.895 -1.459 Td
(0)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(2)Tj
0.502 0 Td
(5)Tj
3.395 0 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
4.254 0 Td
(0)Tj
4.74 0 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
3.529 0 Td
(E)Tj
0.61 0 Td
(l)Tj
0.279 0 Td
(u)Tj
0.502 0 Td
(t)Tj
0.279 0 Td
(i)Tj
0.279 0 Td
(n)Tj
0.502 0 Td
(g)Tj
0.746 0.676 0.668 0.898 k
-21.375 -1.589 Td
(1)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(3)Tj
3.643 0.129 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
4.254 0 Td
(0)Tj
4.74 0 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
0 0 0 0 k
-15.396 -1.589 Td
(1)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(5)Tj
4.146 0.129 Td
(0)Tj
4.254 0 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
4.238 0 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
3.267 0 Td
(W)Tj
0.863 0 Td
(a)Tj
0.443 0 Td
(s)Tj
0.39 0 Td
(h)Tj
0.502 0 Td
(i)Tj
0.279 0 Td
(n)Tj
0.502 0 Td
(g)Tj
0.746 0.676 0.668 0.898 k
-21.642 -1.588 Td
(2)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(0)Tj
4.146 0.129 Td
(0)Tj
4.254 0 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
4.238 0 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
0 0 0 0 k
-15.396 -1.589 Td
(2)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(1)Tj
3.643 0.129 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
4.254 0 Td
(0)Tj
4.74 0 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
0.746 0.676 0.668 0.898 k
-15.396 -1.587 Td
(3)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(0)Tj
3.643 0.128 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
4.254 0 Td
(0)Tj
4.74 0 Td
(1)Tj
0.502 0 Td
(0)Tj
0.502 0 Td
(0)Tj
0 0 0 1 k
/TT1 1 Tf
35 0 0 35 2016.5283 2039.1943 Tm
(Figure 2: )Tj
/TT0 1 Tf
(The LC-MS/MS Setup Used During the Development of this Method. )Tj
/TT1 1 Tf
38.6446 0 0 42.0841 271.6421 785.8662 Tm
(Figure 1:)Tj
/TT0 1 Tf
[( LC Cycle )18(T)37(ime)]TJ
ET
q
808.0799866 0 0 751.1999817 2033.9448242 198.0392761 cm
/Im4 Do
Q
BT
/TT1 1 Tf
34 0 0 34 3568.8691 2074.4878 Tm
(Figure 5A)Tj
36 0 0 36 3725.6709 2074.4878 Tm
(:)Tj
/TT0 1 Tf
34 0 0 34 3737.6592 2074.4878 Tm
( Calibration Curve for Sirolimus \(0.2-200ng/mL\) )Tj
/TT1 1 Tf
21.23 0 Td
(5B: )Tj
/TT0 1 Tf
[(T)111(acrolimus \(0.2-200ng/mL\) )]TJ
/TT1 1 Tf
-20.295 -1.2 Td
(5C: )Tj
/TT0 1 Tf
(Everolimus \(0.2-200ng/mL\) )Tj
/TT1 1 Tf
14.228 0 Td
(5D:)Tj
/TT0 1 Tf
( Cyclosporin \(2-2000ng/mL\))Tj
/TT1 1 Tf
36 0 0 36 2036.7637 988.1831 Tm
(Figure 4: )Tj
/TT0 1 Tf
4.5 0 Td
(EIC Chromatograms of Compounds for QC Level I in Whole Blood)Tj
/TT1 1 Tf
50 0 0 50 4879.9355 1796.2085 Tm
(B)Tj
-14.54 -7.44 Td
(C)Tj
15.5 -0.12 Td
(D)Tj
ET
q
0 2592 5184 -2592 re
W n
0 0 0 0 k
3528.547 1087.296 186 -96 re
f*
3714.547 1087.296 204 -96 re
f*
3918.547 1087.296 132.001 -96 re
f*
4050.548 1059.296 132 -96 re
f*
4182.548 1059.296 132 -96 re
f*
0 0 0 0.8 k
3528.547 991.296 186 -78 re
f*
3714.547 991.296 204 -78 re
f*
3918.547 991.296 132.001 -78 re
f*
4050.548 991.296 132 -78 re
f*
4182.548 991.296 132 -78 re
f*
0 0 0 0 k
3528.547 913.296 186 -46.856 re
f*
3714.547 913.296 204 -46.856 re
f*
3918.547 913.296 132.001 -46.856 re
f*
4050.548 913.296 132 -46.856 re
f*
4182.548 913.296 132 -46.856 re
f*
0 0 0 0.8 k
3528.547 866.439 186 -62.313 re
f*
3714.547 866.439 204 -62.313 re
f*
3918.547 866.439 132.001 -62.313 re
f*
4050.548 866.439 132 -62.313 re
f*
4182.548 866.439 132 -62.313 re
f*
0 0 0 0 k
3528.547 804.127 186 -67.2 re
f*
3714.547 804.127 204 -67.2 re
f*
3918.547 804.127 132.001 -67.2 re
f*
4050.548 804.127 132 -67.2 re
f*
4182.548 804.127 132 -67.2 re
f*
0.746 0.676 0.668 0.898 K
2 w 1 j
q 1 0 0 1 3528.5469 991.2959 cm
0 0 m
786.001 0 l
S
Q
Q
BT
0.746 0.676 0.668 0.898 k
/TT2 1 Tf
27.96 0 0 27.96 3586.5645 1000.9834 Tm
(N)Tj
0.721 0 Td
(a)Tj
0.502 0 Td
(m)Tj
(e)Tj
5.695 0 Td
(M)Tj
(R)Tj
1.665 0 Td
(M)Tj
4.523 0 Td
(Q)Tj
0.777 0 Td
(C)Tj
0.974 0 Td
(1)Tj
2.97 0 Td
(Q)Tj
0.777 0 Td
(C)Tj
0.974 0 Td
(2)Tj
2.97 0 Td
(Q)Tj
0.777 0 Td
(C)Tj
0.975 0 Td
(3)Tj
0 0 0 0 k
-25.292 -2.11 Td
(T)Tj
0.597 0 Td
(a)Tj
0.499 0 Td
(c)Tj
0.443 0 Td
(r)Tj
0.445 0 Td
(o)Tj
0.502 0 Td
(l)Tj
0.279 0 Td
(i)Tj
0.279 0 Td
(m)Tj
0.828 0 Td
(u)Tj
0.558 0 Td
(s)Tj
2.39 0 Td
(8)Tj
0.502 0 Td
(2)Tj
0.502 0 Td
(1)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(6)Tj
0.502 0 Td
(/)Tj
0.279 0 Td
(7)Tj
0.502 0 Td
(6)Tj
0.502 0 Td
(8)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(5)Tj
2.404 -0.022 Td
(3)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(5)Tj
0.502 0 Td
( )Tj
0.249 0 Td
(\()Tj
0.334 0 Td
(9)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(4)Tj
0.502 0 Td
(\))Tj
1.632 0 Td
(1)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(4)Tj
0.502 0 Td
( )Tj
0.249 0 Td
(\()Tj
0.334 0 Td
(2)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(0)Tj
0.502 0 Td
(\))Tj
1.632 0 Td
(2)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(1)Tj
0.502 0 Td
( )Tj
0.249 0 Td
(\()Tj
0.334 0 Td
(0)Tj
0.502 0 Td
(.)Tj
0.249 0 Td
(8)Tj
0.502 0 Td
(\))Tj
0.746 0.676 0.668 0.898 k
-25.754 -2.211 Td
(S)Tj
0.557 0 Td
(i)Tj
0.279 0 Td
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