AMNIOTIC FLUID  Polyhydramnios – volume greater than 1200 ml


Happens when the fetus fails to begin swallowing the amniotic fluid
Transcribed by: DAPNolasco | December 2022
resulting in excess accumulation of it

Secondarily associated with:
Amniotic fluid is present in AMNION, a membranous sac that  Cardiac arrhythmias
surrounds the fetus.  Congenital infections
 Chromosomal abnormalities
Primary functions:  Oligohydramnios – volume less than 800 ml

Provide a protective cushion for the fetus 
Increased fetal swallowing, urinary tract deformities, and

Allows fetal movement membrane leakage are possible cause

Stabilize temp to protect the fetus from extreme temperature

Associated with:
changes  Congenital malformations

Permit proper lung development  Premature rupture of amniotic membranes
 Umbilical cord compression  decelerated heart rate and
fetal death
AMNION is:
 Hemolytic disease of the Newborn:

Metabolically active Rh (-) mother and Rh (+) fetus

Involved in the exchanges of water and chemicals between the
fluid, fetus, and maternal circulation

Produces peptides, growth factors, and cytokines

Color and Appearance

 Colorless
 May exhibit slight to moderate turbidity from cellular
debris, particularly in later stages of fetal development
 MECONIUM
o Newborn’s first bowel movement formed in the
intestine of from fetal intestinal secretions and
swallowed amniotic fluid
o Dark green, mucus-like material
o May be present as a result of fetal distress
o Fetal aspiration of meconium during fetal swallowing
is a concern when increased amounts are present in
fluid
o Very dark red brown fluid is associated with fetal
death

CHEMICAL COMPOSITION

Placenta is the ultimate source of amniotic fluid water and
solutes.

It has similar composition to maternal plasma

Contains small amount of sloughed fetal cells and biochemical
substances produced by fetus that can be tested to determine
health and maturity; also for cytogenetic analysis

Fetal cells in amniotic fluid indicate genetic material of the fetus and the
biochemical substances that the fetus has produced. These cells can be
separated from the fluid. Karyotyping, fluorescence in situ hybridization
(FISH), fluorescent mapping spectral karyotyping (SKY), and DNA
testing. Biochemical substances produced by the fetus can be analyzed
by thin-layer chromatography (TLC) to evaluate the health of the fetus.

Chemical composition of amniotic fluid changes when
VOLUME production of fetal urine begins
Regulated by a balance between: 
Measurement of amniotic fluid creatinine has been used to

production of fetal urine and lung fluid and the absorption from determine fetal age.
fetal swallowing

intramembranous flow (the absorption of amniotic fluid water How to differentiate maternal urine from amniotic fluid?
and solutes into the fetal vascular system) Sometimes, instead of puncturing the amniotic sac, the maternal
bladder is the one that is punctured. Chemical analysis of
 1st trimester: approx. 35ml of amniotic fluid is derived primarily creatinine, urea/BUN, glucose, and protein aids in the
from the maternal circulation differentiation.
 After first trimester: FETAL URINE is the major contributor of 
Creatinine and BUN are lower in amniotic fluid. Creatinine
amniotic fluid does not exceed 3.5mg/dl and urea does not exceed
 At the time that fetal urine production occurs, fetal swallowing of amniotic 30mg/dl in amniotic fluid
fluid begins and regulates increase in fluid from the fetal urine. The fetus 
Measurement of glucose and protein by a reagent strip is a
swallows amniotic fluid, absorbed thru GIT, reexcreted by the kidneys from
less reliable indicator
the blood into fetal urine and back into amniotic fluid. 
Presence of glucose, protein or both is associated closely
 Latter third to half of pregnancy: the fetus secretes a volume of
with amniotic fluid
lung liquid necessary to expand the lungs with growth
 3rd trimester amount increases in quantity throughout pregnancy Analyte Amniotic Fluid Maternal Urine
reaching a peak of approximately 800-1200 ml Creatinine <3.5 mg/dl >10 mg/dl
 it will gradually decrease prior to delivery BUN <30 mg/dl >300 mg/dl
 SPECIMEN HANDLING AND PROCESSING
Fern Test Fluid for Fetal Lung Should be placed in ice (transport) and
- Test used to evaluate the premature rupture of the Maturity (FLM) kept refrigerated prior
membranes (PROM). In here, vaginal fluid specimen is
spread on a glass slide and allowed to completely air dry at Low speed centrifugation is required
room temperature and then observed microscopically. or filtration must be performed to
- The presence of fern like crystals due to protein and prevent loss of phospholipids
sodium chloride content is positive screen for amniotic fluid Sx for bilirubin Must be immediately protected from
testing light
Sx for cytogenetic Must be processed aseptically,
studies/microbial maintained in RT or body temp to
studies prolong life of cells
All fluid for Should be separated from cellular
chemical testing elements and debris ASAP to prevent
cellular metabolism or disintegration

LABORATORY TESTS FOR FETAL DISTRESS

Test for Fetal Hemolytic Disease (HDN)
AMNIOCENTESIS
 Optical density (OD)
It is the procedure used to collect amniotic fluid thru needle 
Read at wavelength of 365 nm to 550 nm
aspiration into the amniotic sac. The procedure performed most 
Normal: Highest absorption at 365 nm
frequently is the transabdominal amniocentesis. Vagina
: decreased linearity at 550 nm
amniocentesis could also be performed however, this method 
If there is HDN: increased absorbance at 450 nm
carries a greater risk of infection.
 (maximum absorbance of bilirubin)
Maximum of 30 ml of amniotic fluid is collected in sterile 
Absorbances are plotted in a liley graph
syringes


Interpretation:
First 2 or 3 ml are discarded as it may be contaminated with
 Zone 1: normal or mildly affected
maternal blood, tissue fluid, and cells
  Zone 2: moderate hemolysis or moderately affected
Specimens should be transferred to sterile plastic containers Requires close monitoring anticipating early delivery or
and taken immediately to the laboratory exchange transfusion upon delivery

Fluid for bilirubin analysis in cases of HDN must be protected  Zone 3: severe hemolytic disease
from light at all times. Intervention thru induction of labor or intrauterine
exchange transfusion is considered
Amniotic fluid analysis is done to: 
Precautions:

Detect fetal abnormalities  Specimens must not be contaminated with

Assess fetal health meconium, cellular fluid and hgb because they

Detect fetal age interfere with spectroanalysis

Monitor pulmonary maturity  Maximum absorbance of oxyhgb occurs at 410 nm
and can interfere with bilirubin absorption peak
Indications for Amniocentesis (at 15-18 weeks AOG)  Remedy: extraction with chloroform

Mother’s age of 35 or older at delivery  Do not expose specimen to light

Family history of chromosome abnormalities such as trisomy 21

Parents carry an abnormal chromosome rearrangement Test for Neural Tube Defects

Earlier pregnancy or child with birth defect  Spina bifida and anencephaly

Parent is carrier of a metabolic disorder  Detected by MSAFP blood test, high resolution ultrasound,

Family history of genetic diseases such as sickle cell disease, amniocentesis
Tay-Sachs disease, hemophilia, muscular dystrophy, sickle  Screening test: measure Alpha-fetoprotein (AFP) N.V= 2 MoM
cell anemia, Huntington chorea, and cystic fibrosis  Confirmatory test: Acetylcholinesterase (AChE)

Elevated maternal serum alpha fetoprotein

Abnormal triple marker screening test  Spina bifida

Previous child with neural tube disorder such as spina bifida, or 
Condition where one or more vertebrae fails to develop completely,
ventral wall defects (gastroschisis) leaving a portion of the spinal cord unprotected

Three or more miscarriages

Strongly linked with deficiency in folate (folic acid) in the diet
especially in pregnancy
Indications for Amniocentesis (at 20-42 weeks AOG)

Fetal lung maturity

Fetal distress

HDN caused by Rh blood type incompatibility

Infection
 Anencephaly
Collection and analysis are recommended when the following 
Most of the infant’s brain is missing because it doesn’t develop
screening blood tests yield abnormal results: 
They can’t survive or either is stillborn or dies within few days

Triple screening tests such as: AFP, hCG, and Estriol

Quadruple screening test: AFP, hCG, estriol and inhibin A

REMINDERS TO CONSIDER!

After the 14th Amniocentesis is generally safe to perform

week AOG Test for Fetal Lung Maturity
At approx. 16 Fluid for chromosome analysis is usually
 Respiratory Distress Syndrome
weeks AOG collected 
The most frequent complication of early delivery
Near the end of Tests for intrauterine growth retardation 7 most common cause of morbidity and mortality in premature
 th
2nd trimester are performed infants
Tests for fetal distress and maturity are 
Caused by insufficiency of lung surfactant production and structural
3rd trimester
performed immaturity of fetal lungs
1. Lecithin/Sphingomyelin Ratio (L/S Ratio)

Usually measured by thin layer chromatography

Normal ratio: greater than or equal to 2.0

Note: can’t be done on sx contaminated with blood or meconium

Sphingomyelin
Lipid constantly produced starting after 26 weeks AOG
Lecithin
Primary component of surfactants (phospholipids, neural lipids,
proteins) that makes up alveolar lining and account for alveolar stability.
It is produced at relatively low and constant rate until 35th week AOG
 increase production to stabilize fetal lung alveoli

2. Phosphatidyl Glycerol

PG is also essential for adequate lung maturity, detected after 35 th
week AOG

Production parallels to lecithin but delayed in maternal diabetes

TLC lung profile must include lecithin, sphingomyelin, and PG to
provide accurate measurement of FLM

3. Lamellar Bodies

Densely packed layers of phospholipids representing a storage
form of pulmonary surfactant

Secreted by Type II pneumocytes of fetal lung at about 24 weeks
AOG

Provides surfanct

As fetal lung matures, production increase is reflected by increase
phospholipids and L/S ratio

4. Foam Stability Index


Mechanical screening test to determine presence of lung surface
lipids

Still in use because it can be performed bedside or in the
laboratory

Shows good correlation with L/S ratio and tests for PG

Cannot be performed on contaminated samples  falsely mature
index