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The Journal of International Medical Research
2008; 36: 163 – 170
H GOKER1, IC HAZNEDAROGLU1, S ERCETIN2, S KIRAZLI1, U AKMAN1, Y OZTURK2 AND 1Department of Haematology, Hacettepe University Medical School, Ankara, Turkey; 2Trend Ankaferd Blood Stopper®
levels were decreased following the
standardized mixture of five plants, has
addition of ABS, in parallel with the
been used historically as a haemostatic
prolonged thrombin time. Total protein,
agent but its mechanism of action
albumin, and globulin levels decreased
remains unknown. This study
after the addition of ABS. Our findings
investigated the in vitro effects of ABS on
suggest that ABS stimulates the
haemostatic parameters. When added to
formation of an encapsulated protein
plasma or serum, ABS induced the very
network that provides focal points for
rapid formation of a protein network and
erythrocyte aggregation. ABS has the
erythrocyte aggregation. The levels of
therapeutic potential to be used for the
coagulation factors II, V, VII, VIII, IX, X,
management of haemorrhage and this
XI, and XIII were not affected by ABS.
agent should be investigated further in
Plasma fibrinogen activity and antigen
clinical trials.
KEY WORDS: ANKAFERD BLOOD STOPPER® (ABS); THYMUS VULGARIS; GLYCYRRHIZA GLABRA; VITIS
VINIFERA; ALPINIA OFFICINARUM; URTICA DIOICA; HERBAL REMEDIES; HAEMORRHAGE; HAEMOSTASIS
Introduction
Ankaferd Blood Stopper® (ABS) is a unique folkloric medicinal plant extract, which has historically been used in Turkish traditional haemostatic actions of ABS. We studied the effect of ABS on the status of the principal plants Thymus vulgaris, Glycyrrhiza glabra, Vitis vinifera, Alpinia officinarum and Urtica dioica. Each of these plants has some effect platelets and blood cells) in vitro using routine haemostatic laboratory tests. Since angiogenesis, cellular proliferation, vascular dynamics and cell mediators,1 – 6 however, and mortality, the identification of a novel, effective haemostatic agent might improve H Goker, IC Haznedaroglu, S Ercetin et al.
Haemostatic effects of Ankaferd Blood Stopper®
the management of bleeding in a wide range ABS/950 µl plasma, 10% (2/20) comprising of patients from all disciplines of clinical comprising 150 µl ABS/850 µl plasma, andso on up to a 50% dilution (10/20) Materials and methods
comprising 500 µl ABS/500 µl plasma. All dilutions were left for no longer than 15 internationally accepted guidelines: EN ISO min before carrying out the tests. Each test evaluation of medical devices’; and EN ISO approximately pH 7.35 was used for testing evaluation of medical devices – Selection of (www.iso.org). This investigation was based on the principle of testing the effect of different ABS/plasma dilutions on distinct Asnières sur Seine, France) according to the descriptive study and, hence, no statistical coagulation factor activities were calculated from the calibration standards of the STA-REvolution® haemostasis device.
ANKAFERD BLOOD STOPPER®
The sample of ABS (Ankaferd Blood
MORPHOLOGICAL EVALUATIONS
Morphological evaluations and microscopic Trend Teknoloji Ilaç AS, Istanbul, Turkey examinations of peripheral blood cells were used in this study was lot number 0806002 carried out in the Morphology Laboratory, Department of Haematology, HacettepeUniversity Medical School and used an HAEMATOLOGICAL TESTS
Ankaferd Blood Stopper® was diluted with by adding 50 µl of ABS solution to a drop of range of dilutions from 0% to 50% in order to study the in vitro effects of ABS on several routine haematological parameters,including individual coagulation factors (II, BIOCHEMICAL TESTS
V, VII, VIII, IX, X, XI and XIII), prothrombin Ankaferd Blood Stopper® was diluted with time (international normalized ratio) (PT total protein, albumin and globulin levels were determined using routine biochemical (TT), D-dimer test, platelet aggregation test dilutions were: 5% (1/20) comprising 50 µl H Goker, IC Haznedaroglu, S Ercetin et al.
Haemostatic effects of Ankaferd Blood Stopper®
rapidly (< 1 s) in the presence of ABS, thereby Ankaferd Blood Stopper® induced very rapid participating in protein network formation (< 1 s) formation of a protein network in the factors (II, V, VII, VIII, IX, X, XI, and XIII), PT (INR), aPTT, fibrinogen, TT, D-dimer, total protein, albumin and globulin after theaddition of ABS to fresh normal plasma or Discussion
In this study, we observed that the addition Coagulation factors II, V, VII, VIII, IX, X, XI of ABS to normal plasma and serum resulted and XIII were not affected by the addition of in the very rapid (< 1 s) formation of a ABS to plasma. Plasma fibrinogen activity protein network. We believe that this ABS- mg/dl to < 30 mg/dl, in parallel with TT haemostatic components in vitro failed to proceed because of the presence of the ABS- induced protein network (data not shown). decreased after the addition of ABS to fresh serum (Table 2). Blood cells, particularly FIGURE 1: The appearance of fresh normal serum before (A) and after (B) the addition
of Ankaferd Blood Stopper® (ABS). ABS resulted in very rapid (< 1 s) formation of a
protein network (B), the microscopic structure of which is shown in C; original
magnification ×10.
H Goker, IC Haznedaroglu, S Ercetin et al.
Haemostatic effects of Ankaferd Blood Stopper®
H Goker, IC Haznedaroglu, S Ercetin et al.
Haemostatic effects of Ankaferd Blood Stopper®
TABLE 2:
Levels of total protein, albumin and globulin at a 5/20 dilution of Ankaferd Blood
Stopper® (ABS) with human serum compared with control
Total protein (g/dl)
Albumin (g/dl)
Globulin (g/dl)
a’Test’ comprises a dilution of ABS/pooled plasma of 5/20. In the ‘Control’ the ABS was replaced by OwrenKoller buffer.
FIGURE 2: Light photomicrograph showing erythrocytes before (A; original
magnification ×10) and after (B; original magnification ×20) the addition of Ankaferd
Blood Stopper® (ABS) to fresh whole blood. ABS resulted in very rapid (< 1 s)
aggregation of erythrocytes
proteins, mainly fibrinogen, and indicated that ABS could affect both fibrinogen and erythrocyte aggregation. However, in plasma other proteins possibly via agglutination of interacts with fibrinogen as well as other formation is related to the functions of blood blood proteins. Since individual clotting factors (coagulation factors II, V, VII, VIII, IX, mechanism of action for ABS appears to be H Goker, IC Haznedaroglu, S Ercetin et al.
Haemostatic effects of Ankaferd Blood Stopper®
agglutination. Blood cells (erythrocytes and platelets) also aggregated and participated increases in the presence of fibrinogen.16 The observations suggest that the ABS-induced formation of the protein network affected the hyperaggregation in the thrombotic process without unequally affecting any individual clotting factor. ABS might, therefore, be effective both in individuals with normal haemostatic parameters and in patients with greater extent than normal fibrinogen.
secondary haemostasis, including patients coagulation. The basic mechanism of action influence on the aggregation process than for ABS appears to be the formation of an on plasma viscosity.18 Plasma cell disorders, encapsulated protein network that provides such as multiple myeloma, represent ‘disease focal points for erythrocyte aggregation.
models’ for the interactions of neoplastic Exposure to ABS seems to provide a tissue paraprotein, erythrocyte aggregation and TT haemostatic process without affecting any fibrinogen activity and fibrinogen antigen prolongation after the addition of ABS.
advantage over other haemostatically-active Likewise, biochemical tests also revealed that total protein, albumin, and globulin levels decreased after the addition of ABS. Hence, Some plant extracts can affect the normal rheological properties of erythrocytes and others penetrate erythrocyte membranes by modifying lipid–protein interactions.10,11 There are close relationships between protein concentrations, polymer type and erythrocyte standardized mixture of the plants T. vulgaris, G. glabra, V. vinifera, A. officinarum and U. dioica, each of which has some effect on haematological and vascular parameters, hypercoagulability.9 Intensified erythrocyte and cellular proliferation.1 – 6 For example, aggregation locally produced in individual G. glabra inhibits angiogenesis, decreases rheological properties of blood flow in the neovascularization.4 G. glabra also has anti- inflammatory, anti-thrombin, anti-platelet, H Goker, IC Haznedaroglu, S Ercetin et al.
Haemostatic effects of Ankaferd Blood Stopper®
anti-oxidant, anti-atherosclerotic, and anti- dynamics and cellular mediators should be tumour activities.4 T. vulgaris has been shown further investigated, however, in order to to exhibit varying levels of anti-oxidant activity, which may help to prevent in vivo pathological states, including neoplastic disorders, infectious diseases, premature peroxidation, associated with atherosclerosis.2 aging, atherosclerosis and diabetes.
Inoculation experiments on detached leaves of V. vinifera exhibited enhanced resistance and mortality in any clinical setting. ABS, a towards pathogens.5,6 V. vinifera also has traditional folkloric medicinal plant extract, is a novel effective haemostatic agent that effects.19,20 A. officinarum inhibits nitric oxide has the therapeutic potential to be used in hoped that clinical trials with this promising U. dioica can produce hypotensive responses through a vasorelaxation effect mediated by development of a new drug that is active in opening of potassium channels, andthrough a negative inotropic action.3 The Acknowledgement
combination of these plants in ABS appears We should like to thank Aykut Koroglu for to provide a unique composition for a tissue carrying out the microscopic examinations.
oxygenation and a physiological haemostaticprocess without disturbing the levels of any Conflicts of interest
individual clotting factor. The effects of ABS Ankaferd Blood Stopper® is a traditional folkloric medicinal plant extract that has angiogenesis, cellular proliferation, vascular • Received for publication 5 August 2007 • Accepted subject to revision 16 August 2007 Copyright 2008 Field House Publishing LLP References
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Dr Ibrahim C Haznedaroglu
Department of Haematology, Hacettepe University Medical School, Sihhiye, Ankara TR-

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