PMID- 6314841
OWN - NLM
STAT- MEDLINE
DCOM- 19831220
LR  - 20190628
IS  - 0003-2697 (Print)
IS  - 0003-2697 (Linking)
VI  - 133
IP  - 1
DP  - 1983 Aug
TI  - The extraction of leukotrienes (LTC4, LTD4, and LTE4) from tissue fluids: the 
      metabolism of these mediators during IgE-dependent hypersensitivity reactions in 
      lung.
PG  - 30-9
AB  - The metabolites of arachidonic acid known as the leukotrienes are a class of 
      lipid mediators which have potent and diverse biological effects in pulmonary 
      tissue. Leukotrienes C, D, and E (LTC4, LTD4, and LTE4) are known to be principal 
      mediators of immunoglobulin E (IgE)-mediated hypersensitivity reactions in lung 
      tissue. It is therefore important to develop reliable and quantitative isolation 
      techniques for estimating levels of these mediators in tissue. In this study, 
      LTC4, LTD4, and LTE4 were separated from other arachidonate metabolites by 
      organic extraction procedures. 5-Hydroxyeicosatetraeonic acid and leukotriene B4 
      extract efficiently into the organic layer of aqueous:ether or aqueous:chloroform 
      extractions, whereas arachidonate metabolites containing conjugated peptides 
      (e.g., LTC4, LTD4, and LTE4) failed to extract into these organic solvents. An 
      extraction step was therefore developed that affords quantitative extraction of 
      LTC4, LTD4, and LTE4 into the organic phase of an isopropanol:ether:H2O mixture. 
      This step is the key for a two-step extraction method that isolates histamine, 
      LTC4, LTD4, and LTE4 with a recovery of 100, 85, 75, and 57%, respectively. One 
      advantage of this separation procedure for obtaining these mediators by organic 
      extraction is an ability to expediently process many samples. Furthermore, the 
      leukotriene content of extracted samples can be analyzed using the guinea pig 
      ileum bioassay without interference from vasoamines or platelet-activating 
      factor. These later substances are eliminated from leukotriene-enriched fractions 
      by this extraction process. When histamine and LTC4 were added to supernatant 
      fluids recovered from isolated lung tissue, they were quantitatively recovered 
      using this extraction method.(ABSTRACT TRUNCATED AT 250 WORDS)
FAU - Clancy, R M
AU  - Clancy RM
FAU - Hugli, T E
AU  - Hugli TE
LA  - eng
GR  - HL06692/HL/NHLBI NIH HHS/United States
GR  - HL16411/HL/NHLBI NIH HHS/United States
GR  - HL25658/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - 0 (SRS-A)
RN  - 37341-29-0 (Immunoglobulin E)
RN  - 75715-89-8 (Leukotriene E4)
RN  - 820484N8I3 (Histamine)
SB  - IM
MH  - Animals
MH  - Body Fluids/*analysis
MH  - Chromatography, High Pressure Liquid/methods
MH  - Guinea Pigs
MH  - Histamine/metabolism
MH  - Hypersensitivity/*metabolism
MH  - Immunoglobulin E/physiology
MH  - Leukotriene E4
MH  - Lipid Metabolism
MH  - Lung/*metabolism
MH  - SRS-A/*analogs & derivatives/*metabolism
EDAT- 1983/08/01 00:00
MHDA- 1983/08/01 00:01
CRDT- 1983/08/01 00:00
PHST- 1983/08/01 00:00 [pubmed]
PHST- 1983/08/01 00:01 [medline]
PHST- 1983/08/01 00:00 [entrez]
AID - 0003-2697(83)90218-X [pii]
AID - 10.1016/0003-2697(83)90218-x [doi]
PST - ppublish
SO  - Anal Biochem. 1983 Aug;133(1):30-9. doi: 10.1016/0003-2697(83)90218-x.