Organic Sewage Tracers

© NERC All rights reserved

Organic Sewage Tracers

• Sterols are components of cells & are synthesized by most organisms

• In humans cholesterol is biohydrogenated by intestinal microflora to 5cholestrol

• Other animals also produce 5cholestrol but at much lower concentrations

• Herbiverous animals produce 24-ethyl coprostanol in greater concentrations

12

3

45

67

89

10

1112

13

14 15

16

17

20

21 22

2324 25

26

27

2829

18

19

A B

C D

Sterol structure


Assessment of Faecal Contamination Using Sterol Molecular Markers

Attenborough Ponds Nature: Long Term Effects of River Diversion


Conneries datingAttenborough ponds CN

0

10

20

30

40

50

60

70

80

0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00

Cs-137 concentration (Bq/kg)

Dept

h (c

m)

1 sigma errors shown

1986 ?

1963 ?


TOC vs Sterol Concentration


Identification of Sewage Markers

17.5 20.0 22.5 25.0minutes

2.5

5.0

7.5

10.0

12.5

15.0

GCounts

Target mix

22.2 22.3 22.4 22.5 22.6 22.7 22.8minutes

2.5

5.0

7.5

10.0

12.5

GCounts

01020304050607080

MCounts

0

25

50

75

100

125MCounts

0

25

50

75

100

MCounts

TIC

m/z = 333Cholesterol-d6

m/z = 231Coprostan-3-one

m/z = 329Cholesterol

Resolving co-elution

17.5 20.0 22.5 25.0minutes

10

20

30

40

50GCounts

0

10

20

30

40MCounts

0

5

10

15

20

25MCounts

0

25

50

75

100

MCounts

0

50

100

150

200MCounts

Sediment Sample

coprostanol

m/z 333

m/z 231

m/z 329

est

ron

e

estr

adio

l

5-c

hole

stan

e

copr

osta

nol

5-e

pico

pros

tano

l

5-c

hole

stan

olca

mpe

ster

olst

igm

aste

rol

fuco

ster

ol +

s

itost

erol

5-s

tigm

asta

nol

Environment & Human Health Theme


•Ten of eleven sewage markercompounds were identifiedin the Attenborough cores

•5-cholestane not detected


sterolR=H: cholest-5-en-3 ; (cholesterol); [57-88-5]; mammalsR=CH : 24 ; (campesterol); [474-62-4]; plantsR=C H : 24-ethylcholest-5-en-3 ; ( -sitosterol); [83-46-5, 19044-06-5]; plants

β-olα-methyl-5-cholesten-3β-ol

β-ol β3

2 3

HO

R

5 ...stanolβ-R=H : 5 ; (coprostanol); [360-68-9]

C 24-methyl- ampe ???-??-?C 24-ethyl- stigma

β-cholestan-3β-olR= H : 5 ; (5 c stanol), [ ]R= H : 5 ; (5 stanol); [19466-47-8]

β-cholestan-3β-ol β-β-cholestan-3β-ol β-

2 5

3

HHO

R

5 epi...stanolβ-Intensive microbial reworking of faecal material in waters and sediments in the environment

Intestinalmicrobialreduction

Microbial reduction in the natural environment

R=H: ; ( epicoprostanol); [516-92-7]C5β-cholestan-3α-ol

R= H : (epi- campestanol); [???-??-?]R=C H : - -cholestan- -ol; (epi- stigmastanol); [???-??-?]

55 5

β-β 3α β-2 5 24-ethyl

5β-

3

HHO

R

5 ...stanolα-R=H: 5 ; ( cholestanol); [80-97-7]

C campe camp ???-??-?stigma stigma

-cholestan-3 -olR= H : 5 ; (5 estanol); [ ]R=C H : 5 ; (5 stanol); [???-??-?]

α- stan-3β-ol α-α- stan-3β-ol α-2 5

α β α-5

3

HHO

R

5α-stanone (intermediate)

HO

R

Δ4 stenone (intermediate)-

O

R

5β-stanone (intermediate)R=H: 5β-coprostanone; [601-53-6]

HO

R

Δ5 stenone (intermediate)-

O

R

cholest-5-en-3 -d (cholesterol-2,2,3,4,4,6-d ); [92543-08-3]β-ol6; 6

HD

2

2D

DD

HO

Sterol & Stanol Marker Pathways

Intestinal (5)

Environmental (5)


1. Sitosterol: Clifton and Church = Input from plants & or avian FM2. Stigmastanol: Conneries = Input from ruminants / manure, slurry ?3. Coprostanol: Concs. >0.5ug/g = received appreciable amounts of sewage

1 2 3


HH O

Coprostanol

H O

Cholesterol

Ratios >0.2 indicative of feacal contam ination (Grim alt & A lbaiges, 1990)

HH O

Cholestanol

HH O

Coprostanol

HH O

Coprostanol

1) M easure of sewage contam ination

HH O

Coprostanol

HH O

Epicoprostanol

HH O

Coprostanol

HH O

Epicoprostanol

HH O

Cholestanol

Ratios 0 .7 indicative of feacal contam ination


H HHO

Coprostanol

Epicoprostanol

HHO

Coprostanol

HO

Cholesterol

2) Measure of sewage treatment or age

Ratios 0 to 0.5 sewage old or treated Ratios 1.5 to 3 sewage fresh or untreated (Mudge etal., 1999)

H HHO

Coprostanol

Epicoprostanol

3) Combined 1 & 2

4) Other sterol markers - Terrestiral plants

HO

Presence of sitosterol indicator of terrestrial run-off(may indicte non-point source diffuse pollution from agricultural areas)

HO

HO


1. Coprostanol/Cholesterol: human sewage >0.2 criteria= Only Conneries

2. Coprostanol/(Coprostanol+Cholestanol) human sewage >0.7= Conneries

3. Epicoprostanol/coprostanol:treatment or age in environ. =Church &

Clifton little sewage or high degree of treatment


1.5

1.0

0.5

0.00.5 1.0 1.5 2.0 2.50.0

HH O

Coprostanol

H O

Cholesterol

HH

HO

Cop

rost

anol

Epi

cop

rost

anol

HO

Low sewage, old or treated

High sewage, untreated

Bi-plot of sewage marker compounds


•Ratio 5Cholestanol/Cholesterol:anaerobic process marker

•Conneries possible more reducing environ at 20 to 50 cm ?

Organic Sewage Tracers

Documents

Transcript of Organic Sewage Tracers