The Big Guy sean at BEST.COM
Sat Sep 2 01:36:02 EST 1995

I need help locating cases regarding  accidential or nonaccidential poisoning
involving  phenolphthalein. I am particularily interested in those that
mention the test procedure used for the dectection of the phenolphthalein.

I greatly appreciate your time and assistance with this project!!

Let me introduce myself:
 I  am biochemist located in Milpitas,CA USA. I have a medical technology
as well as a chemical engineering degree.I have been involved in industrial
research for 15+ years. I joined this group to learn more about the
medicinal and chemical compounds found in plants and herbs.

I am doing independent research concerning a personal problem;

 I couldn't and still can't find the answer to this problem in
books,journals.even online searches.

I was desperate and came up with the idea of the computer I have a degree
in medical technology and
chemistry as well as 15 years+ in chemical research. So I went to fellow
scientists,food scientists like
Dr. Wolstad at Oregon State, Dr. Geza Hrazdina at Cornell, biochemists at
Ottawa: Dr. S Perkins and Dr. W. Grant Thompson, both have done studies
involving laxatives. Others have helped me as well: Francis Durst at
Universite Louis Pasteur,  Dr.William L Hasler, Dept of Internal
Medicine.Univ of Michigan Medical School, Andrew Lyon, Clinical Biochemist
Dept. Pathology, Royal Univ. Hospital Saskatoon, SASK,CANADA, and John
Markwell,  Dept. Biochemistry, University of Nebraska

        My research project regarding phenolphthalein and false positives
initially started because this test method used for a test for
phenolphthalein on a young girl diagnosed with Irritable Bowel Syndrome.Due
to persistant diarrhea,36-48 hours before the test was performed she
underwent a colonoscopy and 3 day clean-out consisting of Magnesium citrate
and bisacodyl laxatives. The colonoscopy was basically normal. As well as I
have now found out she was tested on numerous other occasions for
phenolphthalein using the same test method and they were all negative and
she shows no other signs of  long term laxative abuse.All other lab tests
results are not indicative of laxative abuse.

         This patient has medically documented fevers as high as
105,lymphatic infections, T&A involving a great deal of infectious tissue,
ANA 1:80 on 3 occasions, SED rates as high as 48 without diarrhea
present,test proven allergies, history of chronic URI,pharyngitis(sp?),
cervical lymphoid node enlargements and chronic diarrhea and
gastrointestinal problems. The documented family history reveals history of
stomach and gastrointestinal problems.

        The family situation is one of extreme emotional stress due to a
long divorce and custody battle involving this child and not the older
siblings. The child chose to stay in foster care instead of going to live
with the other parent for over 6 months. The child is not in therapy.

        The parent has been examined by psychological experts who have
experience with Munchausen syndrome by proxy cases including expert
testimony to not have Munchausen syndrome by proxy. This parent chose to go
to a jury trial knowing that was the only chance be reunified with the
child. The child's other parent's attorney started the Munchausen theory
about a year earlier after the client didnot get physical custody of the
chlld and very limited visitation.

        I would like to provide you with some additional information that
will answer some of the questions you might have.

        I have done extensive research at over 6 medical libraries.

        The information is not in text books,journals, dissertations and
very little on-line; but I have located and found the resources but they
dated back to 1870-1930's because the test is that old. I have by going to
the internet to very specific groups [experts in each field I need] I have
made contact with a Biochemist, Food scientists, chromatography
researcher,lab medicine, phytopharmacognosy,analytical chemistry and
toxicologist. These experts each conform that as far as they as they are
knowledgeable in their field,my research is factual and reasonable but no
ONE source can confirm it true on all levels. For example, Dr. Wolstad and
Dr. Hrazdina.confirm that the fruits do contain components that can and do
turn pink-red in alkalined solution and the prunus fruits have been proven
to have cathartic properties But they also had no experience with
anthocyanins in an alkaline aenviroment .Antibiotics do cause diarrhea
especially if on long term maintenance for reoccuring infections.The other
pieces belong to the puzzle but they don't all fit together just yet
without one expert to confirm it all. the following is some of the
resources I have used.

I have read the following materials.
PILANDO, L.S. and WROLSTAD, R. E. 1992. Compositional profiles of fruit
juice concentrates and
sweeteners. Food Chem. 44: 19-20 PILANDO, L.S. and WROLSTAD, R. E. 1992.
The effectiveness of pattern recognition, sugar, nonvolatile acid and
(superscript13)C/(superscript 12)C analyses detecting adulteration in
applejuice. J. Food Comp. Anal. 5: 10-24.SKREDE, G., WROLSTAD, R. E. and
ENERSEN, G. 1992. Color stability of strawberry and blackcurrant syrups. J.
Food Sci. 57: 172-177.  ROMMEL, A., WROLSTAD, R. E. and HEATHERBELL, D. A.
1992 Blackberry juice and wine: Processing and storage effects on
anthocyanin composition, color and appearance. J. Food Sci. 57:385-391,
SPANOS, G.A. and WROLSTAD, R. E. 1992. Phenolics of apple, pear,and white
grape juice and their changes with processing and storage--A Review. J.
Agric. Food Chem. 40: 1478-87.

        Ellenhorn Medical Toxicology(1988), 22. Over-the-counter products:
Laboratory,AmJ Hospital Pharm (Jan. !973).30, 26-71 J Am Pharm Assoc.(Mar
1973).3,130-141,155 Baum,Hm.The occurrence of a diphenylisatin in
California prunes.J Am Pharm Assoc 1951;40;3488-9 Mrak EM,Caffeic acid in
prunes and its behavior as a laxative principle.Sci(1935);82:304
Hutchens,Alma R.:Indian Herbalogy of North America pp. 197-198.
Co-pigmentation of anthocyanins under physiological conditions. J. Food
Sci. 43:517. Williams, M. and G. Hrazdina. 1979. Anthocyanins as food
colorants: Effect of pH on the formation of anthocyanin-rutin complexes. J.
Food Sci. 44:66. Chen, Lao-Jer and G.Hrazdina. 1981. Structural aspects of
anthocyanin-flavonoid complex formation and its role in plant color.
Phytochemistry 20:297-303. Chen, Lao-Jer and G.Hrazdina. 1982. Structural
transformation reactions of anthocyanins. Experientia 38, 1030-1032.
Hrazdina, G. 1982.Anthocyanins. In "The flavonoids; Advances in Research
1975-1981". (J. B. Harborne and T. J. Mabry, eds), Chapman and Hall, London
. S. Perkins, Clin. Bichem vol. 26 pp.179-181. 1993.

        Another chemist I know insisted that colormetric chemical analysis
for phenolphthalein's pink-red endpoint just upon the addition of NaOH is a
very specific and very sensitive test.I have a disagreeing opinion and
decided to prove my research. I believe that this is just a simple
acid-base reaction and not very specific or sensitive for phenolphthalein
only.There are too many other variables that can affect the test.

Colormetric chemical analysis for : PHENOLPHTHALEIN
Test is performed on a urine or stool sample.
- Sodium hydroxide [a base ] is added to the sample and the colormetric
reaction will yield a pink-red coloring upon alkalization of the
sample.This suggests the presence of  phenolphthalein in the sample.
Using thin-layer chromatography, both phenolphthalein bisacodyl/hydrolysis
products turn violet when sprayed with sodium hydroxide. Bisacodyl's Rf
relative to phenolphthatein  [100] is 98.
How  sensitive to interfering substances are these tests?

        Aren't there substances with a ring structure and/or phenolic
protons like phenolphthalein has, that the ring will open when exposed to
an alkaline solution  release the phenolic proton?And turn pink-red color?
and could this resulting color look similar to a phenolphthalein endpoint?

        From the research I have found so far, bisacodyl metabolites give a
similar color and rf as phenolphthalein using thin-layer chromatography.
I'm theorizing since thin-layer is a more specific and sensitive test;
bisacodyl and phenolphthalein metabolites are very similar[both
diphenolics] ;and the fact anthraquinones laxatives are very similar in
their chemical structures to bisacodyl and  turn red like phenolphthalein
in NaOH as a false positive reaction for pphenolphthalein. There is a great
possiblity bisacodyl  does cross react and turn red in alkaline solution.

        Do you think that bisacodyl metabolites could cross-react and
produce a red color similar to phenolphthalein in alkalline solution? Their
metabolites are diphenolics? Correct?

   General comments about Phenol - phthaleins

        The characteristic pink phenolphthalein colour developed in
alkaline solution is the result of a reversible structural change creating
a more highly conjugated species ie.  generally an alternating
sequence of double and single bonds resulting in a lowering of the energy
(ie. can be affected by visible rather than UV light) required to cause
electron transitions.   The phenolic group is particularly amenable to such
pH induced changes readily forming a ketonic or enol form which in the case
of "phenolphthaleins" leads to conjugation with the aromatic arms of the
molecule.  I use the plural because many phenols form compounds
structurally analagous to phenolphthalein e.g. cresolphalein,
thymolphthalein etc. are common commercial pH indicators. Indeed a general
qualitative test for the phenolic group relies on a test tube synthesis of
a phenol-phthalein and observation of its colour in alkaline solution.
This colour needn't however be red, for example thymolphthalein is blue in

        Another common reaction of phenols is their propensity to oxidise
in alkaline solution.  These oxidation products are often tarry complexes
which may appear red in low concentrations.  This is not a reversible
reaction however and should not be confused with the pH driven structural
rearrangement type reactions of indicators.


The phenolic groups on bisacodyl are protected (deactivated) by
esterification and this will prevent
the "phenolphthalein" rearrangement in alkaline solution and colour
development.  After hydrolysis however the phenol group is freed and should
therefore undergo an alkaline colour change as indicated in the "Clinical
Paediatrics" reference which you site.  Bisacodyl may become red if left in
NaOH for some time due to hydrolysis- I don't know what the kinetics of
hydrolysis may be.


According to the Merck Index this discolours in light.  I tried adding some
phloroglucinol to NaOH.  There is no apparent immediate reaction, although
being a tri-phenol the possibility of oxidation, albeit slow, cannot be
discounted. Indeed over the course of a couple of hours my alkaline extract
acquired a distinct pinkish-purple colour.

     Natural Anthroquinone derivatives

Again according to Merck, emodin produces a "cherry-red" colour in alkali
hydroxides, sodium carbonate and ammonia.  This could be a problem!  Don't
know about tumeric acid.  However, I did fiddle about with tumeric, the
spice, at home with the kids and found some mild pH activity, I think.
Merck sites curcumin (tumeric yellow) as a pH active natural root colouring
agent (brownish-red (alkaline) - light yellow (acid)!

     I know from my research that anthraquinones laxatives and their
derivatives also produce a red in an alkaline solution like NaOH.

        Anthraquinone derivatives are found and occur chiefly as
glycosides:anthocyanins,flavoids,and carotoids in a variety of plants and
fruits.Anthoquinones are found occurring naturally in many fruits
including:blackberries, raspberries, cherries, plums, apples, grapes,
nectarines,peaches,pineapples and others? These fruits contain caffeic
acid, O-methyltransferase,tumeric acid,emodin,tartaric acid and /or salts.
Could these turn red like phenolphthalein when alkalinized with NaOH??

        Concentrations of phenolics(chlorogenic acid, caffeic acid,
p-coumaric acid, catechin, epicatechin, two procyanidins, phloridzin, rutin
and hydroxymethylfurfural) have been found in apple juice. SPANOS, G.A. and
WROLSTAD, R. E. 1992. Phenolics of apple, pear, and white grape juice and
their changes with processing and storage--A Review. J. Agric. Food Chem.
40: 1478-87.

        The hydrolysis and metabolism products of anthocyanin and
flavonoids' chemical structure - results in the formation of benzoic acid
derivatives(from the B-ring) and phlorglucinal derivatives (from the
A-ring). Could these turn red like phenolphthalein when alkalinized with

Dr. Hrazdina wrote:
>While anthocyanins may be used a general pH indicators, (red in acidic,
>colorless in mildly acidic, purple at neutral and blue in alkaline
>pH),they also form complexes with other compounds where the same color
>variations may occur.
>Good luck with your research!
>Geza Hrazdina

        It is known that humans metabolize phenolphthalein(unconjugated
with the lactose ring open) but also can store its conjugated form as a
diphenylmethane or phenolphthalein glucuride or another form as rhein
If a person was on a restricted liquid only diet and consumed only
fruit/anthocyanins containing liquids:juices- apple berries,grape etc.,
popsicles, grape flavored heavy on the syrup snowcones,ricelyte, jello
could that person consume enough anthocyanins/anthraquinones to cause
diarrhea and have stool sample turn pink-red upon the addition of an
alkaline solution?

Also what if this person also was consuming way above the FDA limit of food
coloring, food additives, and food preservatives in addition to the above
could a stool sample turn pink-red when exposed to an alkaline solution?

This person was given phenergan suppository 18 hours before a pink-red test

This person was prescribed 20 mg bisacodyl,diphenolic laxative by
suppositories 36 which is way above the amount recommended 5 mg  for  a
child 6-12.

Thank you for reading this very long message!
Can you shed some new light on this subject?
I graciously appreciate any time you take with this project and any help
you can give me!

Now the biggest part I left out,this is all being researched because this
child is a member of  my family.I have taken her at her word and have been
involved since day1 and IBS still fits her pattern of illness especially
since there was a great deal of emotional and stress issues involved during
this time period.And due the fact the test was positive only once and
especially under unusual condition and she was prescribed the other
laxatives so close to the time the test was done. SHE WANTS TO GO HOME!

Well now you know what I am researching and why.I hope you don't mind my
complete honesty and will consider assisting me with this matter?

Patty Ames
patty at sean.vip.best.com


       I can provide with addresses,e-mail,or phone numbers if
        you would like to confirm anything included in my message.

More information about the Toxicol mailing list