HPLC method

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amit_chem
amit_chem's picture
HPLC method

Sir,
I am working on Histamine project.
I tried lot of sets of experiment & search the leterature also,but i didnt get satisfactory HPLC metod for Histamine.
Will u pls help me for the same.
Thank You...

rafiqmo7amed
rafiqmo7amed's picture
Column:

Column:
150 X 3.9 4 |xm Novapak C18
Mobile phase:
to pH 6.55 with phosphoric acid. B was MeCNrwater 80:20. A:B 92:8 for 2 min, to 80:20 over
5 min (Waters convex curve 5), to 65:35 over 28 min (Waters concave curve 7), to 50:50 over
10 min, to 0:100 over 21 min, maintain at 0:100 for 11 min, return to initial conditions over
0.5 min, re-equilibrate for 12.5 min.
Gradient. A was DMF:9 mM NaH2PO4 containing 0.16% triethylamine, adjusted
Column temperature:
50
Flow rate: 1
Injection volume:
10
Detector: UV
436

rafiqmo7amed
rafiqmo7amed's picture
 

 

Column:
150 X 3.9 4 |xm Novapak C18
Mobile phase:
to pH 6.55 with phosphoric acid. B was MeCNrwater 80:20. A:B 92:8 for 2 min, to 80:20 over
5 min (Waters convex curve 5), to 65:35 over 28 min (Waters concave curve 7), to 50:50 over
10 min, to 0:100 over 21 min, maintain at 0:100 for 11 min, return to initial conditions over
0.5 min, re-equilibrate for 12.5 min.
Gradient. A was DMF:9 mM NaH2PO4 containing 0.16% triethylamine, adjusted
Column temperature:
50
Flow rate: 1
Injection volume:
10
Detector: UV
 436

rafiqmo7amed
rafiqmo7amed's picture
sorry there is anerror in

sorry there is anerror in formating
MeCN= ACETONITRILEgradient A= was Dimethylformamide : 9 mM NaH2PO4 containing 0.16% triethylamine, adjusted to pH 6.55 with phosphoric acid.
gradient B= was MeCN : water  80:20
A:B    92:8 for 2 min, to 80:20 over 5 min (Waters convex curve 5), to 65:35 over 28 min (Waters concave curve 7), to 50:50 over 10 min, to 0:100 over 21 min, maintain at 0:100 for 11 min, return to initial conditions over 0.5 min, re-equilibrate for 12.5 min

Column temperature:
50
Flow rate: 1
Injection volume:10Detector: UV 436 

rafiqmo7amed
rafiqmo7amed's picture
 

 

Column:
150 X 1.6 3 jxm Spherisorb 3S TG
Mobile phase:
Gradient. MeCN:water 65:35 for 1 min, to 80:20 over 4 min, to 90:10 over 1 min.
Flow rate: 0.8
Injection volume:
10
Detector: UV
 254

rafiqmo7amed
rafiqmo7amed's picture
Column:

Column:
250 X 4 7 |xm 300 A Nucleosil C8
Mobile phase:
EDTA and 5 mM sodium 1-pentanesulfonate or sodium 1-octanesulfonate.)
MeOH:buffer 15:85 (Buffer was 50 mM pH 3.1 NaH2PO4 containing 0.5 mM disodium
Flow rate: 1
Injection volume:
1-2
Detector: UV
210