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Comparative plasma heparin levels after subcutaneous administration of calcium and magnesium heparin
TZROXBOSIS RESEARCH 23; 33-39, 1981 30L9-3848;81/130033-07$02.00/0 Copyright (c) 1981 Pergamon ?ress Ltd. .411rights reserved.
COXPARATIVE PLASMA HEPARIS LEVELS AFTER SUBCUTANEOUS XMINISTRATION OF CALCILM XUD YAGWESIL?1HEPARIN
Gilles FOLLEA, ?4arie-ChristineTRZECIAK, %rc DECUAVA>~E Laboratoire d'Hemobiologie, Institut Pasteur, Faculte Alexis Carrel, 69372 Lyon Cedex 2, France.
(Received 20.2.1981; in revised form 12.5.1981. Accepted by Editor Y. Sultan)
ABSTRACT In a cross-over experiment six normal subjects were given subcutaneously an injection of calcium heparin and another injection of magnesium heparin (from the same batch). Following the administration of the magnesium salt, plasma heparin levels measured by three specific assays were found significantly higher one hour after (~4.05) and lower seven hours after (p
INTRODUCTION During the last few years there was an increased use of subcutaneous heparin either at low dose in the prophylaxis of post-operative deep vein thrombosis (1) or at high dose for therapy of deep vein thrombosis or pulmonary embolism (2,3). At the present time, three kinds of heparin salts are available : sodium, calcium and magnesium heparins. Recently three studies emphasized the role of the salt in the anticoagulant activity of heparin. When heparin is given subcutaneously, the peak plasma levels are lower with the calKEY
WORDS:heparin,
administration and dosage, pharmacodynamics. 33
vo1.23,
x0.1/2
cium salt than vith ihe sodium one (C,5,6). To cur knowledge, only one study compared the plasma heparin levels after subcutaneous injection or’ sodium, calcium and magnesium heparins (7). The results showed no difference, but the heparins used were not prepared from the same batch. The aim of our study was to compare subcutaneous calcium and magnesium heparins, regarding plasma heparin levels after the administration of these two salts. Our overall results indicate significantly different plasma levels between the two kinds of heparin one and seven hours after subcutaneous injection, and suggest a faster absorption rate for magnesium heparin. ?iATERIhLAKD ?fETHODS Reagents - Blood was collected in 0.106 M trisodium citrate (l/9). The time elapsed between venipuncture and centrifugation never exceeded 10 minutes (at room temperature). Platelet poor plasma (PPP) was obtained by centrifugation 4,000 g for 20 minutes at room temperature, and stored at -60°C for no longer than 2 months. Pooled normal plasma (PNP) was obtained in the same conditions from 30 healthy blood donors. - For our first trial, magnesium and calcium heparin were prepared by Biosedra (France) from the same bulk batch of heparin powder, and the identity of heparin was concealed by relabelling them respectively H1 and Ht. These 2 heparins contained respectively 6.24 p.100 magnesium and 10.25 p.100 calcium. For our second trial, commercial calcium and magnesium heparins were obtained from Choay (Calciparine@, batch 742-l) and Biosedra (Cutheparine@, batch 119>, and relabelled respectively H3 and Hb. All four heparins were extracted from hog intestinal mucosa. Molecular weight was estimated by gel chromatography on a column (90 x 2.6 cm) of Sephacryl S 200 with 0.2 M NaCl as eluant and a flow rate of 20 ml/h. Calibration was made with Dextran Tls and Dextran TAO. For the 4 heparins, the mean molecular weight was identical : 15,000 daltons (range 5,000 to 35,000). Isoelectric focusing of heparin in acrylamide gel was performed as described by Nader et al. (8). An ampholyte mixture pH 3-5 was obtained from LKB. There was no difference in the distribution of the bands between the 4 heparins. The French assays stated 24,458
anticoagulant activity of the 4 heparins was assayed according to the Pharmacopeia method (9) and the values quoted were from 4 independent against the same standard for heparin (Diosynth, Netherlands). The potencies were respectively (in IU/ml) : 24,987 (HI), 25,150 (Hz), (Hs) and 26,125 (H4).
Heparin assays Anti-Xa clotting activity was evaluated by the method of Denson and Bonnar (IO) with reagents from Diagnostic Reagents Ltd (Thame, England). AntiXa amidolytic activity was evaluated by the method of Teien et al. (II) with chromogenic substrate S-2222 (Kabi, Sweden) on an automatic device (Isamat, ISA Biologie), and without addition of antithrombin III (AT III). Thrombin calcium clotting time (TCCT) was carried out with the method described by Penner (12), using a human thrombin (Fibrindex, Ortho) at 1 NIH U/ml in 0.05 ?lCaC12. Standard curves were built with pretreatment PPP (anti-Xa clotting assay) or PW (anti-Xa amidolytic assay and TCCT) treated with the corresponding
injected heparin, and all the results were PPP was used to dilute every tested plasma curve. AT III
expressed found out
as of
IV/ml. Pretreatment the range of the
assays
They uere performed with the method of Xbildgaard et al. (13) using the chromogenic substrate S-2238 (Kabi) and an automatic device (Isamat, ISA Biologic) . Standard curves were built :;ith PSP taken as 100 2 activity. Design
of
the
study
Subjects participating to the study were normal adults (Hospital Staff, 25 to 43 years old) and informed consent was obtained from all of them. They refrained from medication IO days before and throughout the study. In a first cross-over trial 6 women were given 2 subcutaneous injections (200 U/kg each, 2 days apart) of calcium (Hz) and magnesium (HI) heparin from the same batch. The order of injection was randomly allocated and only the physician was aware of this order. The injections were made at 8 a.m. by the same nurse. Blood samples were drawn from each subject immediately before injection and In a second identical cross-over trial, 6 1, 3, 5, 7 and 8 hours thereafter. injections (200 other subjects (1 man, 5 women) were given 2 subcutaneous U/kg) of calcium (Ha) and magnesium heparin (H4) from a commercial source. In another randomized study, 3 normal subjects (1 man, 2 women) were given intravenously 5,000 units of calcium (Hz) and magnesium (HI) heparin from Blood samples were drawn before injection and the same batch, 2 days apart. IO, 30, 60, 120 and 180 minutes thereafter.
III
On every blood were measured. For
statistical
sample,
plasma
comparisons,
heparin
a paired
leve 1 (3 specific
test
assays)
and AT
was used.
RESULTS In the first study (calcium and magnesium heparin from the same batch), statistical comparison of the differences between each paired plasma heparin levels obtained with the 3 assays showed significantly higher plasma levels with the magnesium heparin 1 hour after injection, and higher plasma levels with the calcium heparin 7 hours after injection (table I). No difference was observed between the 2 salts 3, 5 and 8 hours after injection. The mean plasma heparin levels (pooled 3 assays) obtained in this first trial are shown in figure I. Comparison of the differences between each paired plasma levels obtained after intra-venous administration of these 2 heparins in 3 subjects gave no difference between the 2 salts. In the second trial (calcium and magnesium heparins from a commercial I source), we found higher plasma heparin levels with the magnesium heparin and higher plasma levels with the and 3 hours after subcutaneous injection, calcium heparin 7 hours after injection (table II). Overall correlation between anti-Xa clotting levels and TCCT was highly significant (r : 0.762, p
Differ?_nces
: ii;
I tion
I
j !
&kti-:
(hours) _ /
*
3jS
1 + . 0;:
-
. 3; 7
3 7 s
+ . 1x5 (.165) +. +.
172 (.I@)
_
*
i-
-
. 013
!
+ . Oh5 (.077)
1
.
. Or;9:: \.3a1, . 015
(.lh9)
097 (.060)
’
OjC
(.089)
(.OY8) j
-
j -
(.:a)
* . 045 (. i 35)
co: !.l@S)
+ . 10 1::::
+ . OS6
/.!29)
(.136)
i+ . 045 055 - . iii; +. (.l47) i (-041) ; (.147)l
H: and HI : respectively calcium and magnesium :: p c 0.05 ; :::: p < 0.01
3
t3SfS
i (.075) / /
(.l84)
?oo;z:
; I
.
(.085) 3
(ICh.1)
-S-,-T _/-.r _
,:;I:i-:‘:2 azidoly_=ic _
033
- 3:
!
026 (.1i5)
heparin.
TABLE II PIean differences (and standard deviation) between plasma heparin levels after subcutaneous injection of comercial calcium and magnesium heparin.
Differences
: H? - H4 (ID/ml)
tion (hours) 1
3 5
. 159 (.123)
-
- . 102 (.166)
-
+.
027
*
(.Oi6) 081
.
8 .
+.
069 (.148)
+ . Oil
*
+ . 024
+.
(.lOY) +.
i
085 (.032)
+ . oi5 (.197)
-
(.IOi)
(.OYO) 7
I
- . 136 I (.C87) /
083
.
H3 and Hb : respectively calcium and magnesium ..\> ,.,.p < 0.01 ; :::::: p -c O.OlJl.
12$:x:: i:.SSl)
.
rJ7y::: (.i32)
055 (.137)
-
Oh? ;.210)
+ . 031 i . :38)
134 (.186)
+.
j + . 044
(.Oj6) j
’
()‘$fj:::: i . 133)
; + . 0'3 .il) (.043) / \.'
heparin.
7
3.
PLASMA
HEPARI
(Ida1
N LEVELS
)
MAGNESIUM .-----’ 1
CALCIUM
HEPARIN HEPARIN
I 5
3 TIME
FIG.
AFTER
I 7
INJECTION
I 8 (HOURS)
1
Comparative plasma heparin levels (mean k S.D. from pooled 3 tests in 6 subjects) after subcutaneous administration of calcium and magnesium heparins from the same batch. highly significant (r : 0.792, p < 0.001) but the clotting method gave significantly higher levels than the amidolytic method (p < 0.001). AT III assays gave values in the normal range for all the blood specimens and we observed no modification of the AT III levels up to 8 hours after heparin injection with the 4 heparins. While no local pain was reported after subcutaneous injection of the 2 calcium heparins, 416 and 3/b subjects experienced pain with the magnesium heparins, respectively in the first and the second study. No local hematoma was seen in all the 12 subjects. DISC'JSSIOY The results of our first trial indicate differences between calcium and magnesium heparin and we are not aware of another similar report comparing subcutaneous calcium and magnesium heparins from the same batch. But the differences we found between subcutaneous commercial calcium and magnesium heparins are at variance with the only study using magnesium heparin (7), in which no difference was noted between the plasma heparin levels after subcutaneous injections of sodium, calcium and magnesium heparins. The use of only one assay (thrombin clotting time) in the cross-over experiment (5 subjects) of Bouvier (7) may probably partly explain the discrepancy between his study and ours.
-
T3.2 &if-’ 2r2nc2s <>‘::air.& i?. :,iir study ‘between :he caiaiu?. and magnesium. heparins from the same batch cannet be explained by a differential clearance plasma heparin levels foilowing intra-venous rate because of the similar administration of the 7 salts. In addition, t1:ere was r.0 difference between * r;eight clstri’bution. Our rethe 4 ‘heparins used regarding their molecular sults suggest a faster absorption rate for magnesium h2parin as compared to some clinicalcium heparin. However, we have as yet no ar.gument to attribute cal relevance to these findings.
Overall correlations between the 3 heparin assays used in this study were highly significant, and no difference was found between the anti-Xa clotting the anti-Xa anidolvtic assav gaire signilevels and the TCCT levels. However, ficantly lower values than the anti-Xa clotting assay, and these discrepancies could not be explained by decreased AT III levels. Such a difference has already be mentionned by others (14,15) and no clear explanation appears to be available. Marciniak and Gockerman (16) reported a progressive reduction in AT III, in patients receiving intra-venous heparin therapy. In this study, we never observed a modification of AT III levels up to 8 ‘hours after a single subcutaneous injection, whatever the heparin used. These findings are in accordance to those of Kakkar et al. (17) who found a decrease in AT III only in intravenously treated patients but not in subcutaneously treated patients. However, controversial results have been presented by Green (18), showing decreased AT III values after 3 days of subcutaneous heparin. Our results tend to indicate that a single dose of subcutaneous heparin produces no change in AT III, as stated by Marciniak and Gockerman (16) for single doses of intravenous heparin. ACKNOWLEDGMENTS We are grateful to Mrs Chantal to Mrs Fabienne Martin for typing McGregor for reviewing the english S-2222 used throughout this study
Huchon for skilful technical assistance the manuscript. We also thank Dr. J.L. manuscript. The chromogenic substrates were a generous gift from Kabi.
and
REFERENCES Present status Low-dose heparin. 25, suppl. 36, 158-178, 1980.
and future
trends
: Scud.
I.
KAKKAR, V.V. J. Hamtiat.
2.
HIRSH, J., O’SULLIVAN, E.F., GALLUS A.S. calcium heparin therapy in the treatment J. Au/~&. I, 15-17, 1970.
3.
BENTLEY, P.G., KAKKAR, V.V., SCULLY, M.F., ?!cGREGOR, I.R., P JONES, x. An objective study of alternative methods of ni:tration. Thrramb. R&i. 18, 177-187, 1980.
4.
JOHNSON, E.A., KIRKWOOD, T.B.L., STIRLING 1 Y., PEREZ-REQUEJO, J.L., INGRAM, J.I.C., BANGHAM,D.R., BROZOVIC, M. Four heparin preparations anti-Xa potentiating effect of heparin after subcutaneous injection. Thhomb. ff~em~d-t. 35, 586-591, 1976.
5. THOHAS, D.P., SAGAR, S., STAMATAKIS, J.D., KAKKAR, V.V. sodium salts
Plasma heparin levels of heparin. Thmb.
after RCA. 9,
et of
al. Evaluation thromboembolic
MAFFEI, F.H.A., administration of 241-248, 1976.
of subcutaneous disease. bicd.
WEBB, P., CHAS, heparin admi-
ERDI, A., calcium and
:
vo1.23,
Xo.l!2
39
6.
LOW, J., BICGS, J.C. Comparative plasma heparin levels after subcutaneous sodium and calcium heparin. Thtcrr;D. H.z??c~~.40, 397-&06, 1978.
7.
BOWIER C.X. Comparison at the clinical level of different types and routes of heparin administration. Stand. J. Ha&q~tcL. 25, suppl. 36, 91100, 1980.
6. NADER, H.S., McDUFFIE, N,M., DIETRICH, C.P. Heparin fractionation b) electrofocusing : presence of 21 components of different molecular weights. 8icch. Bioph. R&A. Ccm. 57, ii@-h93, 197s. 9. Phamacopee Franqaise, 8eme edition, p. 559-562. 10. DEUSON K .W .E ., BO%AR, J. The measurement of heparin. A method based on . Piaih. Hcccmo&z. 30, 471-479, the polentiation of anti-factor Xa. Tko&. 1973. II. TEIEN, A.N., LIE M. Evaluation of an amidolytic heparin assav method : increased sensitivity by adding purified antithrombin III. Tj:d~omb. Rc.4, IO, 399-410, 1977. 12. PENNER, J.A. Experience with a thrombin clotting time assay for measuring heparin activity. Am. 1. c&l. P&d. 61, 645-653, 1974. 13. ABILDGAARD, U., LIE,M., @DEGARD, O.R. Antithrombin (heparin cofactor) assay with "new" chromogenic substrates (S-2238 and chromozym TH) : Thumb. Rcs. II, 549-553, 1977. 14. BARROWCLIFFE, T.W., JOHNSON, E.A., EGGLETO?;,C.A., KEMBALL-COOK, G., THOMAS, D.P. Anticoagulant activities of high and low molecular weight heparin fractions. BCt. J. Haemtitol.41, 573-583, 1979. 15. ANDERSSON, L.O., BARROWCLIFFE, T.W., HOLMER, E., JOHNSON, E.A., SODERSTROM, G. Molecular weight dependency of the heparin potentiated inhibition of thrombin and activated factor X. Effect of heparin neutralization in plasma. Tivwmb. Ru. 15, 531-541, 1979. 16. MARCINIAK, E., COCKERMAN, J.P. Heparin induced decrease in circulating antithrombin III. Lancti 17, 581-584, 1977. 17. KAKKAR, V.V., BENTLEY, P.G., SCULLY, M.F., MCGREGOR, I.R., JONES, N.A.G., WEBB, P.J. Antithrombin III and heparin. Lancti 1, 103-104, 1980. 18. GREEN, P.J. Antithrombin III and subcutaneous heparin. LanCei 1, 374-375, 1980.
COXPARATIVE PLASMA HEPARIS LEVELS AFTER SUBCUTANEOUS XMINISTRATION OF CALCILM XUD YAGWESIL?1HEPARIN
Gilles FOLLEA, ?4arie-ChristineTRZECIAK, %rc DECUAVA>~E Laboratoire d'Hemobiologie, Institut Pasteur, Faculte Alexis Carrel, 69372 Lyon Cedex 2, France.
(Received 20.2.1981; in revised form 12.5.1981. Accepted by Editor Y. Sultan)
ABSTRACT In a cross-over experiment six normal subjects were given subcutaneously an injection of calcium heparin and another injection of magnesium heparin (from the same batch). Following the administration of the magnesium salt, plasma heparin levels measured by three specific assays were found significantly higher one hour after (~4.05) and lower seven hours after (p
INTRODUCTION During the last few years there was an increased use of subcutaneous heparin either at low dose in the prophylaxis of post-operative deep vein thrombosis (1) or at high dose for therapy of deep vein thrombosis or pulmonary embolism (2,3). At the present time, three kinds of heparin salts are available : sodium, calcium and magnesium heparins. Recently three studies emphasized the role of the salt in the anticoagulant activity of heparin. When heparin is given subcutaneously, the peak plasma levels are lower with the calKEY
WORDS:heparin,
administration and dosage, pharmacodynamics. 33
vo1.23,
x0.1/2
cium salt than vith ihe sodium one (C,5,6). To cur knowledge, only one study compared the plasma heparin levels after subcutaneous injection or’ sodium, calcium and magnesium heparins (7). The results showed no difference, but the heparins used were not prepared from the same batch. The aim of our study was to compare subcutaneous calcium and magnesium heparins, regarding plasma heparin levels after the administration of these two salts. Our overall results indicate significantly different plasma levels between the two kinds of heparin one and seven hours after subcutaneous injection, and suggest a faster absorption rate for magnesium heparin. ?iATERIhLAKD ?fETHODS Reagents - Blood was collected in 0.106 M trisodium citrate (l/9). The time elapsed between venipuncture and centrifugation never exceeded 10 minutes (at room temperature). Platelet poor plasma (PPP) was obtained by centrifugation 4,000 g for 20 minutes at room temperature, and stored at -60°C for no longer than 2 months. Pooled normal plasma (PNP) was obtained in the same conditions from 30 healthy blood donors. - For our first trial, magnesium and calcium heparin were prepared by Biosedra (France) from the same bulk batch of heparin powder, and the identity of heparin was concealed by relabelling them respectively H1 and Ht. These 2 heparins contained respectively 6.24 p.100 magnesium and 10.25 p.100 calcium. For our second trial, commercial calcium and magnesium heparins were obtained from Choay (Calciparine@, batch 742-l) and Biosedra (Cutheparine@, batch 119>, and relabelled respectively H3 and Hb. All four heparins were extracted from hog intestinal mucosa. Molecular weight was estimated by gel chromatography on a column (90 x 2.6 cm) of Sephacryl S 200 with 0.2 M NaCl as eluant and a flow rate of 20 ml/h. Calibration was made with Dextran Tls and Dextran TAO. For the 4 heparins, the mean molecular weight was identical : 15,000 daltons (range 5,000 to 35,000). Isoelectric focusing of heparin in acrylamide gel was performed as described by Nader et al. (8). An ampholyte mixture pH 3-5 was obtained from LKB. There was no difference in the distribution of the bands between the 4 heparins. The French assays stated 24,458
anticoagulant activity of the 4 heparins was assayed according to the Pharmacopeia method (9) and the values quoted were from 4 independent against the same standard for heparin (Diosynth, Netherlands). The potencies were respectively (in IU/ml) : 24,987 (HI), 25,150 (Hz), (Hs) and 26,125 (H4).
Heparin assays Anti-Xa clotting activity was evaluated by the method of Denson and Bonnar (IO) with reagents from Diagnostic Reagents Ltd (Thame, England). AntiXa amidolytic activity was evaluated by the method of Teien et al. (II) with chromogenic substrate S-2222 (Kabi, Sweden) on an automatic device (Isamat, ISA Biologie), and without addition of antithrombin III (AT III). Thrombin calcium clotting time (TCCT) was carried out with the method described by Penner (12), using a human thrombin (Fibrindex, Ortho) at 1 NIH U/ml in 0.05 ?lCaC12. Standard curves were built with pretreatment PPP (anti-Xa clotting assay) or PW (anti-Xa amidolytic assay and TCCT) treated with the corresponding
injected heparin, and all the results were PPP was used to dilute every tested plasma curve. AT III
expressed found out
as of
IV/ml. Pretreatment the range of the
assays
They uere performed with the method of Xbildgaard et al. (13) using the chromogenic substrate S-2238 (Kabi) and an automatic device (Isamat, ISA Biologic) . Standard curves were built :;ith PSP taken as 100 2 activity. Design
of
the
study
Subjects participating to the study were normal adults (Hospital Staff, 25 to 43 years old) and informed consent was obtained from all of them. They refrained from medication IO days before and throughout the study. In a first cross-over trial 6 women were given 2 subcutaneous injections (200 U/kg each, 2 days apart) of calcium (Hz) and magnesium (HI) heparin from the same batch. The order of injection was randomly allocated and only the physician was aware of this order. The injections were made at 8 a.m. by the same nurse. Blood samples were drawn from each subject immediately before injection and In a second identical cross-over trial, 6 1, 3, 5, 7 and 8 hours thereafter. injections (200 other subjects (1 man, 5 women) were given 2 subcutaneous U/kg) of calcium (Ha) and magnesium heparin (H4) from a commercial source. In another randomized study, 3 normal subjects (1 man, 2 women) were given intravenously 5,000 units of calcium (Hz) and magnesium (HI) heparin from Blood samples were drawn before injection and the same batch, 2 days apart. IO, 30, 60, 120 and 180 minutes thereafter.
III
On every blood were measured. For
statistical
sample,
plasma
comparisons,
heparin
a paired
leve 1 (3 specific
test
assays)
and AT
was used.
RESULTS In the first study (calcium and magnesium heparin from the same batch), statistical comparison of the differences between each paired plasma heparin levels obtained with the 3 assays showed significantly higher plasma levels with the magnesium heparin 1 hour after injection, and higher plasma levels with the calcium heparin 7 hours after injection (table I). No difference was observed between the 2 salts 3, 5 and 8 hours after injection. The mean plasma heparin levels (pooled 3 assays) obtained in this first trial are shown in figure I. Comparison of the differences between each paired plasma levels obtained after intra-venous administration of these 2 heparins in 3 subjects gave no difference between the 2 salts. In the second trial (calcium and magnesium heparins from a commercial I source), we found higher plasma heparin levels with the magnesium heparin and higher plasma levels with the and 3 hours after subcutaneous injection, calcium heparin 7 hours after injection (table II). Overall correlation between anti-Xa clotting levels and TCCT was highly significant (r : 0.762, p
Differ?_nces
: ii;
I tion
I
j !
&kti-:
(hours) _ /
*
3jS
1 + . 0;:
-
. 3; 7
3 7 s
+ . 1x5 (.165) +. +.
172 (.I@)
_
*
i-
-
. 013
!
+ . Oh5 (.077)
1
.
. Or;9:: \.3a1, . 015
(.lh9)
097 (.060)
’
OjC
(.089)
(.OY8) j
-
j -
(.:a)
* . 045 (. i 35)
co: !.l@S)
+ . 10 1::::
+ . OS6
/.!29)
(.136)
i+ . 045 055 - . iii; +. (.l47) i (-041) ; (.147)l
H: and HI : respectively calcium and magnesium :: p c 0.05 ; :::: p < 0.01
3
t3SfS
i (.075) / /
(.l84)
?oo;z:
; I
.
(.085) 3
(ICh.1)
-S-,-T _/-.r _
,:;I:i-:‘:2 azidoly_=ic _
033
- 3:
!
026 (.1i5)
heparin.
TABLE II PIean differences (and standard deviation) between plasma heparin levels after subcutaneous injection of comercial calcium and magnesium heparin.
Differences
: H? - H4 (ID/ml)
tion (hours) 1
3 5
. 159 (.123)
-
- . 102 (.166)
-
+.
027
*
(.Oi6) 081
.
8 .
+.
069 (.148)
+ . Oil
*
+ . 024
+.
(.lOY) +.
i
085 (.032)
+ . oi5 (.197)
-
(.IOi)
(.OYO) 7
I
- . 136 I (.C87) /
083
.
H3 and Hb : respectively calcium and magnesium ..\> ,.,.p < 0.01 ; :::::: p -c O.OlJl.
12$:x:: i:.SSl)
.
rJ7y::: (.i32)
055 (.137)
-
Oh? ;.210)
+ . 031 i . :38)
134 (.186)
+.
j + . 044
(.Oj6) j
’
()‘$fj:::: i . 133)
; + . 0'3 .il) (.043) / \.'
heparin.
7
3.
PLASMA
HEPARI
(Ida1
N LEVELS
)
MAGNESIUM .-----’ 1
CALCIUM
HEPARIN HEPARIN
I 5
3 TIME
FIG.
AFTER
I 7
INJECTION
I 8 (HOURS)
1
Comparative plasma heparin levels (mean k S.D. from pooled 3 tests in 6 subjects) after subcutaneous administration of calcium and magnesium heparins from the same batch. highly significant (r : 0.792, p < 0.001) but the clotting method gave significantly higher levels than the amidolytic method (p < 0.001). AT III assays gave values in the normal range for all the blood specimens and we observed no modification of the AT III levels up to 8 hours after heparin injection with the 4 heparins. While no local pain was reported after subcutaneous injection of the 2 calcium heparins, 416 and 3/b subjects experienced pain with the magnesium heparins, respectively in the first and the second study. No local hematoma was seen in all the 12 subjects. DISC'JSSIOY The results of our first trial indicate differences between calcium and magnesium heparin and we are not aware of another similar report comparing subcutaneous calcium and magnesium heparins from the same batch. But the differences we found between subcutaneous commercial calcium and magnesium heparins are at variance with the only study using magnesium heparin (7), in which no difference was noted between the plasma heparin levels after subcutaneous injections of sodium, calcium and magnesium heparins. The use of only one assay (thrombin clotting time) in the cross-over experiment (5 subjects) of Bouvier (7) may probably partly explain the discrepancy between his study and ours.
-
T3.2 &if-’ 2r2nc2s <>‘::air.& i?. :,iir study ‘between :he caiaiu?. and magnesium. heparins from the same batch cannet be explained by a differential clearance plasma heparin levels foilowing intra-venous rate because of the similar administration of the 7 salts. In addition, t1:ere was r.0 difference between * r;eight clstri’bution. Our rethe 4 ‘heparins used regarding their molecular sults suggest a faster absorption rate for magnesium h2parin as compared to some clinicalcium heparin. However, we have as yet no ar.gument to attribute cal relevance to these findings.
Overall correlations between the 3 heparin assays used in this study were highly significant, and no difference was found between the anti-Xa clotting the anti-Xa anidolvtic assav gaire signilevels and the TCCT levels. However, ficantly lower values than the anti-Xa clotting assay, and these discrepancies could not be explained by decreased AT III levels. Such a difference has already be mentionned by others (14,15) and no clear explanation appears to be available. Marciniak and Gockerman (16) reported a progressive reduction in AT III, in patients receiving intra-venous heparin therapy. In this study, we never observed a modification of AT III levels up to 8 ‘hours after a single subcutaneous injection, whatever the heparin used. These findings are in accordance to those of Kakkar et al. (17) who found a decrease in AT III only in intravenously treated patients but not in subcutaneously treated patients. However, controversial results have been presented by Green (18), showing decreased AT III values after 3 days of subcutaneous heparin. Our results tend to indicate that a single dose of subcutaneous heparin produces no change in AT III, as stated by Marciniak and Gockerman (16) for single doses of intravenous heparin. ACKNOWLEDGMENTS We are grateful to Mrs Chantal to Mrs Fabienne Martin for typing McGregor for reviewing the english S-2222 used throughout this study
Huchon for skilful technical assistance the manuscript. We also thank Dr. J.L. manuscript. The chromogenic substrates were a generous gift from Kabi.
and
REFERENCES Present status Low-dose heparin. 25, suppl. 36, 158-178, 1980.
and future
trends
: Scud.
I.
KAKKAR, V.V. J. Hamtiat.
2.
HIRSH, J., O’SULLIVAN, E.F., GALLUS A.S. calcium heparin therapy in the treatment J. Au/~&. I, 15-17, 1970.
3.
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