PMID- 20156415
OWN - NLM
STAT- MEDLINE
DCOM- 20100629
LR  - 20211203
IS  - 1096-0309 (Electronic)
IS  - 0003-2697 (Linking)
VI  - 401
IP  - 1
DP  - 2010 Jun 1
TI  - Reversed-phase high-performance liquid chromatography method for simultaneous 
      analysis of two liposome-formulated short interfering RNA duplexes.
PG  - 61-7
LID - 10.1016/j.ab.2010.02.012 [doi]
AB  - Gene silencing induced by short interfering RNA (siRNA) has proven to be useful 
      in genomic research and has great potential for therapeutic applications; 
      however, siRNAs are not readily bioavailable. Cationic liposomes offer effective 
      protection of drug product from nucleases and enable distribution to desired 
      target organs. The amount of siRNA in the formulation must be determined 
      accurately. We have developed a stability-indicating, ion-pair, reversed-phase 
      high-performance liquid chromatography method to separate and accurately 
      quantitate two siRNA duplexes in a liposome without sample pretreatment. The 
      gradient mobile phase system consisted of 385mM hexafluoro-2-propanol, 14.5mM 
      triethylamine, and 5% methanol (mobile phase A) and 385mM hexafluoro-2-propanol, 
      14.5mM triethylamine, and 90% methanol (mobile phase B). The column used was an 
      XBridge C18 column (50x2.1mm i.d., 2.5microm particle size), and separation was 
      performed at 60 degrees C. Quantitation was achieved with ultraviolet (UV) 
      detection at 260nm. Linearity was established for the single strands of both 
      siRNA duplexes for concentrations ranging from 10 to 110microg/ml. Accuracy of 
      the method was determined by replicate analysis (n=5) at four concentrations 
      (R(2)>0.996 and relative standard deviations [RSDs] of 1-4%). The use of an 
      ion-pairing reagent that is compatible with mass spectrometry detection makes 
      this method amenable to liquid chromatography-mass spectrometry (LC-MS) impurity 
      profiling.
CI  - Copyright 2010 Elsevier Inc. All rights reserved.
FAU - Murugaiah, Veeravagu
AU  - Murugaiah V
AD  - Alnylam Pharmaceuticals, Cambridge, MA 02142, USA. mmurugaiah@alnylam.com
FAU - Zedalis, William
AU  - Zedalis W
FAU - Lavine, Gary
AU  - Lavine G
FAU - Charisse, Klaus
AU  - Charisse K
FAU - Manoharan, Muthiah
AU  - Manoharan M
LA  - eng
PT  - Journal Article
DEP - 20100213
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - 0 (Liposomes)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (Vascular Endothelial Growth Factor A)
RN  - EC 3.6.4.4 (Kinesins)
SB  - IM
MH  - Chromatography, High Pressure Liquid/*methods
MH  - Chromatography, Reverse-Phase/*methods
MH  - Kinesins/genetics
MH  - Liposomes/*chemistry
MH  - RNA, Small Interfering/analysis/*chemistry
MH  - Vascular Endothelial Growth Factor A/genetics
EDAT- 2010/02/17 06:00
MHDA- 2010/06/30 06:00
CRDT- 2010/02/17 06:00
PHST- 2009/11/18 00:00 [received]
PHST- 2010/02/05 00:00 [revised]
PHST- 2010/02/09 00:00 [accepted]
PHST- 2010/02/17 06:00 [entrez]
PHST- 2010/02/17 06:00 [pubmed]
PHST- 2010/06/30 06:00 [medline]
AID - S0003-2697(10)00097-7 [pii]
AID - 10.1016/j.ab.2010.02.012 [doi]
PST - ppublish
SO  - Anal Biochem. 2010 Jun 1;401(1):61-7. doi: 10.1016/j.ab.2010.02.012. Epub 2010 
      Feb 13.