Clinical Chemistry 43:11
2155–2163 (1997)
Disappearance of human chorionic gonadotropin
and its a- and b-subunits after term pregnancy
Juha Korhonen,* Henrik Alfthan, Pekka Ylöstalo, Johannes Veldhuis,1 and
Ulf-Håkan Stenman
We have used high-specificity and precision immunoflu-
orometric assays to measure the elimination half-times of
human chorionic gonadotropin (hCG), hCGa, and hCGb in
serum over 21 days after delivery in six women with term
pregnancies. Baseline concentrations and half-times were
calculated with the use of a curve-fitting algorithm for mul-
tiexponential decay. In contrast to the two-component model,
a three-component exponential function with baseline pro-
vided a fit for which predicted values could not be distin-
guished from the observed values by analysis of variance.
Median half-times were 3.6, 18.0, and 53.0 h for hCG; 1.0, 23.4,
and 194 h for hCGb; and 0.6, 6.2, and 21.9 h for hCGa. The
mean ratio of hCGa to hCG decreased rapidly from 36.9% to
3.3% on day 3; thereafter it increased to 64.3% 21 days after
delivery because of a higher baseline concentration of hCGa.
hCGb had the slowest total elimination rate, and the ratio of
hCGb to hCG in serum increased from 0.8% before delivery
to 26.7% after 21 days. If the metabolism of hCG and hCGb is
similar in patients with trophoblastic disease, the ratio of
hCGb to hCG must be evaluated with caution in samples
taken several days after initiating therapy. We conclude that
the disappearance of hCGb from plasma is slower than
previously recognized and that the ratios of hCGb or hCGa to
intact hCG vary as a function of postpartum time. Such
information may be important in clinical studies of preg-
nancy disorders.
Human chorionic gonadotropin (hCG)2 is a heterodimer
composed of two highly glycosylated subunits, called a
Department of Obstetrics and Gynecology and Department of Clinical
Chemistry, Helsinki University Central Hospital, Haartmaninkatu 2, FIN-
00290 Helsinki, Finland.
1
Department of Internal Medicine, University of Virginia Health Sciences
Center, and National Science Foundation Center for Biological Timing, Char-
lottesville, VA 22908.
* Author for correspondence. Fax 358-9-4714801; e-mail ulf-hakan,
stenman@huch.fi.
2
Nonstandard abbreviations: hCG, human chorionic gonadotropin; hCGa
and hCGb, highly glycosylated subunits of hCG; AUC, area under the curve.
Received January 22, 1997; revision accepted July 3, 1997.
2155
Endocrinology and
Metabolism
and b, that are noncovalently joined. The a-subunits of all
glycoprotein hormones of pituitary origin, including fol-
licle-stimulating hormone, luteinizing hormone, and thy-
roid-stimulating hormone, are virtually identical, but the
b-subunits are different and thus confer biological speci-
ficity of the hormones.
In early pregnancy the concentrations of hCG in serum
start to increase 7–11 days after ovulation, corresponding
to 21–25 days after the last menstrual period [1, 2]. After
in vitro fertilization and embryo transfer, an increase of
serum hCG can be observed 9 days after ovum retrieval,
corresponding to 7 days after embryo transfer [3]. The
increase is exponential, with a doubling time of 1.5 days
during the first 6 weeks [4]. Serum hCG reaches peak
concentrations of ;100 000 IU/L (in relation to the First
International Reference Preparation) at 8 –10 weeks after
the last menstrual period. The concentrations start to
decrease after week 12 and stay fairly constant at about
;30 000 IU/L from the 20th week until term [5].
In addition to hCG, serum and urine from pregnant
women and patients with trophoblastic disease contain
free a-subunits (hCGa) and b-subunits (hCGb). The pro-
file of the serum concentrations of hCGb during preg-
nancy resembles that of hCG, but the concentrations are
lower. During gestation, the molar ratio of hCGb to hCG
is 1.5– 4% in early pregnancy and decreases to 0.2–1%
after the 10th week [3, 6]. In patients with benign tropho-
blastic disease, the ratio is similar to that in pregnancy,
whereas higher ratios are observed in trophoblastic can-
cer. Thus the ratio may aid in differentiating between
malignant and benign trophoblastic tumors [7–10].
The concentrations of hCGa increase throughout preg-
nancy from ,1 mg/L (69 pmol/L) [11] to 100 –300 mg/L
(6900 –20 700 pmol/L) in the third trimester [12, 13]. The
hCGa to hCG ratio is ,10% during the first trimester and
increases to 30 – 60% at term [6].
When hCG is injected into humans, it reportedly has a
biphasic disappearance curve with an initial fast half-time
of ;5 h and a slow one of 24 –32 h [14 –16]. Injected hCGa
and hCGb are cleared from circulation much more rap-
idly than hCG [16, 17]. hCGa has a rapid half-time of 13
2156 Korhonen et al.: Disappearance of hCG after term pregnancy
min and a slow one of 76 min. The corresponding times
for hCGb are 41 and 236 min, respectively [18, 19].
Serial hCG estimations are used for detecting pregnan-
cy-related disorders such as spontaneous abortion and
ectopic pregnancy as well as following-up patients with
ectopic pregnancy. In ectopic pregnancy patients selected
for expectant management, decreasing concentrations
usually indicate spontaneous resolution, but one-third of
these still require surgery within 1–24 days [20]. Thus a
decreasing hCG concentration alone is not a reliable
indicator for spontaneous resolution of an ectopic preg-
nancy.
Because hCGa and hCGb have been reported to have
much shorter half-times than hCG, they might more
rapidly reflect changes in placental function such as an
impending abortion. Changes in the ratio of the subunits
to intact hCG potentially could be used to evaluate
trophoblastic activity in pregnancy-related disorders, e.g.,
a low subunit to hCG ratio might indicate the cessation of
hCG production because the ratio may be expected to
decrease rapidly when the production ceases such as
during an abortion or after delivery.
Materials and Methods
patients
Six women with term pregnancies were included in the
study, which was approved by the local institutional
review committee. One of the patients had a twin preg-
nancy, one had gestational diabetes (non-insulin-depen-
dent glucose intolerance of pregnancy), and one had
diabetes mellitus (insulin-dependent diabetes with ne-
phropathy). The mean age was 37 years (range 32– 48
years) and mean gestational age 38 6 4 weeks (range 37 1
3 to 39 1 5 weeks). In singleton pregnancies the mean
birth weight was 3862 g (range 3270 – 4360 g) and the
mean placental weight 678 g (range 600 –770 g). In the
twin pregnancy the birth weights were 2970 g and 3325 g
and placental weights 560 g and 685 g, respectively.
The first blood sample was drawn from a cannula in-
serted in the right cubital vein 15 min before an elective
caesarean section, performed under spinal anesthesia. Infu-
sion of saline solution was started in the opposite arm to
stabilize the hemodynamics during anesthesia before the
beginning of spinal anesthesia. The infusion was continued
in some individuals, depending on the blood pressure. At
the time of the first sample, ;500 mL of saline solution had
been infused and at the time of delivery ;1000 mL. The
effect of saline infusion on the serum concentration of hCG
and its subunits and consequently on the half-times was
analyzed in one case (patient 5 in Table 3), in which blood
samples were drawn before fluid infusion and after infusion
of 1 L of saline. After delivery, blood samples were drawn at
5, 10, 15, 20, 30, 40, 60, 120, 180, 360, and 720 min during the
first day and every 24 h thereafter for 6 days. Additional
blood samples were obtained from all patients 14 to 16 days
after delivery and from 4 of those patients 21 days after
delivery. Serum samples were stored at 220 °C before assay.
immunoassays
Serum hCG, hCGb, and hCGa were determined by time-
resolved immunofluorometric assays (DELFIA®, Wallac).
The detection limit for hCG was 0.3 IU/L. The assays for
hCGb and hCGa are newly developed time-resolved
immunofluorometric assays and are described in detail
elsewhere. The detection limit for hCGb was 0.3 pmol/L
(0.01 mg/L) (conversion factor: 1 mg/L 5 45.5 pmol/L)
and that for hCGa 2.8 pmol/L (0.04 mg/L) (conversion
factor: 1 mg/L 5 69 pmol/L). Cross-reaction of hCG in the
subunit assays was studied after separation of hCG and
the subunits in pregnancy serum [3]. On a molar basis, the
cross-reaction of hCG in the hCGb assay was 0.05% and
that in the hCGa assay ,0.1%. The upper reference limits
of hCG, hCGb, and hCGa in nonpregnant premenopausal
women were 2.9 IU/L, 1.6 pmol/L [21], and 31 pmol/L,
respectively [H. Alfthan, unpublished finding]. The as-
says measure equally intact and nicked forms of hCG and
hCGb [H. Alfthan, unpublished finding].
calibrator preparations and conversion factors
The hCG assay was calibrated against the WHO 3rd
International Standard (75/569), 1 mg corresponding to
9286 IU, and hCGb and hCGa against the 1st Interna-
tional Standard, 75/551 and 75/569, respectively. For
conversion of IU to molar units, molecular masses of
36 700, 22 000, and 14 500 Da were used for hCG, hCGb,
and hCGa, respectively [22]. The conversion factor of
hCG to SI unit was 1 IU/L 5 2.93 pmol/L.
calculations and statistical analysis
The ratio of the subunits (hCGa or hCGb) to total hCG
was calculated on the basis of the molar concentration of
each subunit relative to the concentration of subunit 1
hCG. Percentiles, means, SDs, and 95% confidence limits
for the concentrations of hCG, hCGb, and hCGa and the
ratios of the concentrations to the initial values were
calculated for each time point. The individual subject’s
concentration values were used for calculations of base-
line concentrations and half-times for hCG, hCGb, and
hCGa with the use of a curve-fitting algorithm for mul-
tiexponential decay (Microsoft Excel®). An iterative ap-
proach was used to calculate the combination of basal
concentration and two or three exponential functions
providing the best fit of the observed concentration val-
ues. The algorithm minimizes the fitted variance, which is
the sum of the squares of the differences between the
calculated curve and the observed values.
The mathematical model for the decay curve is defined
as
OC z e
n
f~t! 5 2R i z t
1A (1)
i
i51
where Ci is a coefficient (decay amplitude) and Ri is a rate
constant (min21). t is the time after delivery, and A is the
baseline concentration. By choosing n between 1 and 3,
 Clinical Chemistry 43, No. 11, 1997
sums of up to 3 exponential terms can be used [23].
Untransformed concentration data over time are fit, such
that a triphasic decay curve is defined as
f~t! 5 C 1 z e ~2R 1t! 1 C 2 z e ~2R 2t! 1 C 3 z e ~2R 3t! 1 A
and a biphasic decay curve is defined as
f~t! 5 C 1 z e ~2R 1t! 1 C 2 z e ~2R 2t! 1 A
The half-times are calculated as t1/2 5 (ln2/Ri). The
integrated contribution of each exponential component to
the overall disappearance curves of hCG, hCGb, and
hCGa was calculated on the basis of the area under the
curve (AUC) by
Ci 1 A
AUC 5 2 z @~e 2R i z t ! 2 1#
Ri
Significant differences between triphasic and biphasic
half-time fits were evaluated by comparing the fitted
variances (above) via F ratio testing with Bonferroni/
Dunn correction. We estimated whether the time course
over which the measurements were made affected the
half-times by analyzing time intervals of 0 – 6 days, 0 –1
day, 0 –12 h, and 0 – 6 h. The longest half-times were not
calculated when the time interval was shorter than the
longest half-time component.
Results
The mean serum concentration of hCG before delivery
was 32 275 IU/L (94 566 pmol/L) (range 1631– 84 995
IU/L). The corresponding value for hCGb was 729
pmol/L (range 48 –2198 pmol/L) and for hCGa 35 862
pmol/L (range 17 432– 68 145 pmol/L). The mean concen-
trations of hCG, hCGb, and hCGa are shown in Figs. 1–3,
and the ratios of hCGb and hCGa to total hCG and the
ratios of hCGa to hCGb are shown in Table 1.
A three-component exponential function gave the best
Fig. 1. Observed disappearance of hCG (mean 1 SD) and estimated triphasic exponential half-times of hCG and their combined curve.
2157
fit to the observed hCG decay curves (Fig. 1). In contrast
to a biphasic model, this resulted in very small deviations
from the observed values, and the differences between
triphasic and biphasic models and between the biphasic
(2)
model and the observed values were significant (Table 2).
The curve for hCGb was also optimally fit to a three-
component model (Fig. 2), with significant differences
(3) from the biphasic model (Table 2). The calculated biphasic
curve of hCGb fell below the observed values during the
first hour after delivery, rose above the observed values
during the next 12 h, and also deviated thereafter. The
curve for hCGa was also best fit to a three-component
model (Fig. 3). It fit to a two-component model only when
values from days 0 –2 were included in the calculations.
When days 3– 6, 14, and 21 values were also included, the
(4)
biphasic model gave a poor fit. There were no significant
differences between calculated values of the triphasic
half-time model and the observed values of hCGa,
whereas calculated values of the biphasic model differed
significantly from the observed ones and those of the
triphasic model by analysis of variance (Table 2). The
half-times were virtually identical when calculated for
shorter time intervals.
The median half-time of the most rapid component of
hCG was ;6 times longer than that of hCGa and 4 times
longer than that of hCGb (Table 3). The median half-time
of the second component of hCG was 30% shorter than
that for hCGb, but for hCGa the second component was 3
times shorter. The median half-time of the third compo-
nent of hCGb was almost fourfold that of hCG, and for
hCGa it was half of that for hCG (Table 3). The algorithm
used was set to calculate the baseline concentrations of
hCG and its subunits limiting the highest possible value
to the upper reference limit of nonpregnant premeno-
pausal women. Without this limitation two patients
would have had baseline concentrations for hCG of 3.5
2158 Korhonen et al.: Disappearance of hCG after term pregnancy

Table 1. Ratios (percentage) of hCG subunit to total hCG and ratio of hCGa to hCGb after delivery.
Mean (SD) ratio

Time after delivery n hCGb/(hCGb 1 hCG), % hCGa/(hCGa 1 hCG), % hCGa/hCGb

0 min 6 0.8 (0.2) 36.9 (21.7) 110.9 (116.2)
5 min 5 0.8 (0.2) 38.8 (23.2) 103.5 (102.6)
10 min 6 0.8 (0.2) 34.2 (22.2) 97.4 (102.2)
15 min 5 0.8 (0.2) 35.6 (23.5) 95.0 (102.3)
20 min 6 0.7 (0.2) 31.7 (21.7) 90.2 (96.1)
30 min 6 0.7 (0.2) 29.6 (21.3) 82.2 (86.6)
40 min 6 0.7 (0.1) 27.8 (21.4) 75.3 (80.9)
60 min 6 0.7 (0.1) 23.5 (18.8) 59.5 (60.8)
120 min 6 0.6 (0.1) 17.2 (15.6) 43.8 (44.7)
180 min 6 0.6 (0.1) 14.7 (13.1) 34.1 (30.9)
360 min 6 0.6 (0.1) 11.9 (11.4) 25.0 (23.6)
720 min 6 0.7 (0.1) 9.2 (9.5) 15.6 (15.1)
1 day 6 0.9 (0.1) 5.5 (5.3) 6.9 (6.4)
2 days 6 1.2 (0.2) 3.4 (3.1) 3.0 (2.5)
3 days 6 1.5 (0.3) 3.3 (2.3) 2.4 (1.6)
4 days 6 1.7 (0.3) 4.1 (2.7) 2.5 (1.6)
5 days 5 2.1 (0.3) 5.2 (3.2) 2.8 (1.7)
6 days 5 2.8 (0.5) 7.0 (4.5) 2.8 (1.2)
14 days 4 15.8 (7.4) 45.1 (19.7) 5.0 (1.9)
21 days 4 26.7 (7.6) 64.3 (20.6) 6.7 (4.7)

and 5.2 IU/L, two patients baseline concentrations for
hCGb of 2.8 and 2.5 pmol/L, and one patient a baseline
concentration for hCGa of 34.2 pmol/L.
In one patient (number 5 in Table 3) the serum concen-
trations of hCG, hCGb, and hCGa were measured before
and after induction of spinal anesthesia and infusion of 1
L of saline. The infusion lowered the serum concentra-
tions by ;22%. When the postinfusion concentrations
were used as initial values, the calculated half-times
increased, but the median effect was small (2.2%, range
0 –24%).
The mean concentrations of hCG, hCGb, and hCGa on
day 21 after delivery were 2.6 IU/L (7.6 pmol/L) (range
0.7– 4.3 IU/L), 3.2 pmol/L (range 1.2–5.1 pmol/L), and 16
pmol/L (range 9.3–25.0 pmol/L), respectively. hCGb had
the slowest total elimination rate, and decreased to a
mean 6 SD concentration of 1.3% 6 1.0% of the initial
value after 14 days and 0.9% 6 0.8% after 21 days. The
hCG concentration decreased to 0.05% 6 0.02% of the
initial value at 14 days and 0.02% 6 0.01% within 21 days.
hCGa reached a concentration of 0.07% 6 0.03% within 14
days, decreasing only slightly thereafter to 0.05% 6 0.03%
of the initial value on day 21 (Fig. 4).
The contributions of the various components to the
disappearance curves were calculated on the basis of
AUCs. For hCG, the fastest component represented 13.8%
of all hCG and two slower ones were 68.8% and 17.4%,
respectively. For hCGb, the corresponding numbers were
3.1%, 78.1%, and 18.9%, and for hCGa they were 23.9%,
59.7%, and 16.4%, respectively (Table 4). hCG contained
much more of the most rapid component than hCGb.
Therefore, the total disappearance of hCGb was clearly
slower than that of hCG (Fig. 4), and after a transient
decrease during the first hour after delivery, the ratio of
hCGb to hCG increased gradually from 0.8% (range
0.6 –1.0%) before delivery to 15.8% (range 7.7–28.3%) after
14 days and 26.7% (range 20.6 –36.9%) after 21 days (Fig.
5).
hCGa had the highest proportion of the rapid compo-
nent and the lowest proportion of the slow component,
explaining its most rapid total disappearance (Table 4 and
Fig. 4). The mean ratio of hCGa to hCG before delivery
was 36.9% (range 15.3–78.5%). Initially the ratio decreased
rapidly with a nadir of 3.3% (range 1.4 –7.9%) on day 3,
but thereafter it increased to 64.3% (range 42.5– 83.5%) 21
days after delivery (Fig. 6). This resulted from a constant

Table 2. Significance of the fitted variances of triphasic vs biphasic half-time curves of hCG, hCGb, and hCGa.
P-valuea

Form of hCG Triphasic vs observed Biphasic vs observed Triphasic vs biphasic

hCG 0.9138 ,0.0001 ,0.0001
hCGb 0.8531 ,0.0001 ,0.0001
hCGa 0.7063 ,0.0001 ,0.0001
a
Comparisons by analysis of variance are significant when the corresponding P is ,0.0167.
 Clinical Chemistry 43, No. 11, 1997 2159

Fig. 2. Observed disappearance of hCGb (mean 1 SD) and estimated triphasic exponential half-times of hCGb and their predicted overall decay
curve.

background concentration of hCGa with decreasing hCG
concentrations.
Discussion
With the use of highly sensitive and specific immunoflu-
orometric assays, we found in this study that the disap-
pearance of endogenous hCGb from plasma after delivery
is much slower than that observed in studies with the use
of RIA and purified hCGb infused intravenously [18, 19].
Various possible explanations exist for this. Chemical
differences, mainly in the carbohydrate side chains [24 –
26], have been shown to exist between circulating hCGb
and that purified from pregnancy urine, which has been
used for injection [18, 19]. The experimental setting in the
present study is not directly comparable with that after
infusion of purified hCGb; i.e., the half-time was mea-
sured for hCGb over a course of 3 weeks, whereas in
earlier reports the time span has been only hours or a few
days. However, for comparison, we also calculated half-
times for shorter time frames that were comparable with
those in earlier studies. This did not affect the results
substantially. After pregnancy, some release of small
amounts of hCGb (and hCG) sequestered in tissues could
cause an apparent increase in half-time. Another possibil-
ity is that trophoblastic cells remaining in the body
continue to produce hCGb and very little or no hCG.
Trophoblasts persist in the lungs for extended periods of
time after pregnancy [27], but the mass of these cells is
very small in comparison with that of the placenta.
Therefore, any production of hCG and hCGb by persist-
ing trophoblastic cells would not contribute significantly
to the serum concentrations observed the first week after

Fig. 3. Observed disappearance of hCGa (mean 1 SD) and estimated triphasic exponential half-times of hCGa and their algebraically summed
curve.
2160 Korhonen et al.: Disappearance of hCG after term pregnancy

delivery, when the difference in half-times was already

Table 3. Individual baselines and triphasic half-times (h) of
apparent in an increasing ratio of hCGb to hCG (Fig. 5).
hCG, hCGb, and hCGa after delivery.
Dissociation into subunits of the hCG remaining in
Half-times, h
circulation could also affect the estimated half-time of
Base Rapid (I) Medium (II) Slow (III) hCGb. This mechanism could be important if the disap-
hCG, IU pearance of hCGb were more rapid than that of hCG, but
Patient 1 2.90 2.08 17.26 50.88 the opposite was actually true. Intact hCG is quite stable,
Patient 2a 2.90 4.88 18.64 48.72 whereas nicked hCG dissociates more rapidly. Especially
Patient 3 2.35 3.71 19.16 64.10 in trophoblastic disease [10, 28], nicking may increase the
Patient 4 0.21 3.46 16.66 55.02 dissociation of hCG into subunits, which may contribute
Patient 5 0.66 2.04 12.82 38.11
to a high ratio of hCGb to hCG. Long incubation times
Patient 6 1.22 6.17 25.32 60.86
during the assay of hCGb can cause dissociation of hCG,
Mean 1.71 3.73 18.31 52.95
thus increasing the apparent concentration of hCGb in the
SD 1.17 1.61 4.10 9.31
sample. However, with the incubation times used in the
Median 1.79 3.58 17.95 52.95
present assay for hCGb, this effect is negligible [3].
hCGb, pmol/L Although all these mechanisms could increase the half-
Patient 1 1.60 1.12 23.50 191.56 time of hCGb, they probably do not explain why hCGb in
Patient 2a 1.59 0.79 26.83 196.60 all subjects studied disappeared substantially more
Patient 3 1.60 1.08 23.26 134.78
slowly than hCG. Therefore, other explanations need to be
Patient 4 0.65 0.93 21.81 214.86
considered.
Patient 5 0.00 0.80 22.26 462.12
The most likely explanation for the longer half-time of
Patient 6 0.93 1.25 24.33 102.64
hCGb (vs intact hCG) is that hCGb circulating in plasma
Mean 1.06 0.99 23.67 217.09
SD 0.66 0.19 1.79 127.27
differs from that isolated by dissociation of urinary hCG
Median 1.26 1.01 23.38 194.08 into subunits. hCG in urine is known to be less glycosy-
lated than that in serum [24]. The carbohydrate composi-
hCGa, pmol/L
tion, and especially the presence of terminal sialic acid, is
Patient 1 5.01 0.70 8.79 126.26
known to affect the in vivo half-time of hCG [26]. Fur-
Patient 2a 25.61 0.54 4.74 21.38
thermore, the b-chain of both hCG and hCGb in crude
Patient 3 9.35 0.85 8.49 100.12
urinary hCG preparations has been found to be partially
Patient 4 31.00 0.35 3.28 15.25
Patient 5 10.88 0.63 6.64 17.69
cleaved or nicked between residues 47 and 48 [25, 29]. In
Patient 6 22.96 0.62 5.67 22.45 addition, under the potentially harsh chemical conditions
Mean 17.47 0.62 6.27 50.53 required to dissociate the subunits, denaturation and
SD 10.43 0.17 2.15 49.31 appearance of components with shortened half-times
Median 16.92 0.63 6.16 21.92 could occur. The hCG heterodimer is unusual in that it is
a
Patient with diabetes mellitus (white F).
held together by a loop of the b-chain embracing the

Fig. 4. Disappearance of hCG, hCGb, and hCGa after delivery.

The values are given as the proportion (mean 1 SD) of the concentration before delivery. The SD bars not visible are too small to be seen at this scale.
 Clinical Chemistry 43, No. 11, 1997 2161

Table 4. Proportions of the various components of hCG, hCGa, and hCGb estimated on the basis of the AUC of each
component.
AUCa

Form of hCG Rapid (I) Medium (II) Slow (III) Total

hCG 0.48 (13.8) 2.41 (68.8) 0.61 (17.4) 3.50 (100)
hCGb 0.14 (3.1) 3.45 (78.3) 0.82 (18.6) 4.41 (100)
hCGa 0.13 (23.9) 0.32 (59.7) 0.09 (16.4) 0.54 (100)
a
Given in parentheses is the percent of the total.

a-chain [30]. Disrupting the dimer in vitro might therefore
change the structure of hCGb in comparison with the
circulating form, which probably never has been involved
in heterodimer formation. Free hCGa in serum does not
reassociate with hCGb [31, 32].
An increased ratio of hCGb to hCG, in most studies
.10%, has been observed in trophoblastic cancer, and this
has been used to differentiate between malignant and
benign trophoblastic disease [7–9, 33]. In the present
study, ratios .10% were observed in 5 of the 6 patients
studied 14 days after delivery, and in all 4 patients
studied 21 days after delivery. If the metabolism of hCG

Fig. 5. Changes in the ratio of hCGb to total hCG (mean 1 SD) after delivery.
The values are given as percentages.

Fig. 6. The ratio of hCGa to total hCG (mean 1 SD) after delivery.
The values are given as percentages.
2162 Korhonen et al.: Disappearance of hCG after term pregnancy
and hCGb is similar in patients with trophoblastic dis-
ease, our findings suggest that the ratio of hCGb to hCG
must be evaluated with caution in samples taken several
days after initiation of therapy. An increased ratio of
hCGb to hCG has actually been observed several weeks
after treatment of trophoblastic disease [34, 35].
The two most rapid components of hCG had half-times
similar to those observed for hCG injected into humans,
i.e., 3.6 and 18 h as compared with 5 and 24 –36 h,
respectively [14 –16]. In an earlier study three components
with half-times of 15, 27, and 168 h were estimated for
hCG after abortion [36]. The two latter half-times support
our calculations about the third component with the
half-time of several days (median 53 h for hCG). This
component represented only 17% of total hCG. Therefore,
it may not be detectable after injection unless a large
amount of hCG is injected, or an ultrasensitive assay
method is used, or very prolonged observations are
carried out. However, it is possible that this component
represents hCG produced by residual, gradually dying
trophoblasts or that is less abundant in urinary hCG than
in plasma. The half-times of the two most rapid compo-
nents of disappearance of hCG, hCGb, and hCGa were
similar in all the patients studied, but there was more
individual variation in the half-times of the longest com-
ponent, i.e., from 38 to 64 h for hCG, 103 to 462 h for
hCGb, and 15 to 126 h for hCGa. Renal clearance accounts
for 20% of the total disposal of hCG after injection of
purified hCG preparations [19]. The slightly impaired
kidney function in our patient with diabetes mellitus
(patient 2 in Table 3) seemed to have no effect on the
half-times of hCG and its subunits.
The algorithm used for calculation of half-times in the
present study was based on the principles described in the
EXPFIT program [23]. A two-component model has been
used in most earlier studies to calculate disappearance
half-times of hCG and its subunits. However, the fit of a
two-component model was unsatisfactory for hCG,
hCGb, and hCGa, whereas a three-component model
with baseline yielded a statistically preferred fit. In con-
trast to the two-component model, three exponentials
with baseline provided a fit for which predicted values
could not be distinguished from the observed values by
analysis of variance. The baseline concentrations obtained
with the algorithm were in most cases well within the
range of the reference values for nonpregnant premeno-
pausal women (Table 3) [21]. However, when the fol-
low-up time is insufficient for analysis of the baseline, it
can be restricted in the algorithm.
In conclusion, we have developed a three-component
exponential model with a baseline for calculation of
half-times of hCG and its subunits. Disappearance of
endogenous hCGb from plasma after delivery is slower
than previously observed, and the ratios of hCGb or
hCGa to intact hCG vary as a function of postpartum
time. If the metabolism of hCG and hCGb is similar in
patients with trophoblastic disease, the ratio of hCGb to
hCG must be evaluated with caution in samples taken
several days after initiation of therapy. Ratios of hCGb to
hCG .10%, which are indicative for chorionic cancer,
were observed in all patients 21 days after delivery.
However, this needs to be evaluated in patients with
trophoblastic disease. Additional studies will also reveal
whether the half-times are similar in early pregnancy and
whether this can be used to diagnose pregnancy-related
disorders.
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