～（99）Tc～m-DEMDP的制备及生物分布研究
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摘要
亚甲基二膦酸二乙酯(DEMDP)是当今核医学领域应用最广泛的骨显像剂亚甲
基二膦酸(MDP)的类似物。文献中有关DEMDP的合成及其金属螯合物的研究已
有一些报道，但是，对于DEMDP的99Tcm标记及标记物的生物分布尚未见报道。
本文对由合成MDP的母液中提取出来的DEMDP进行了元素分析、核磁共
振、红外光谱、质谱分析，从而对其结构进行了表征。
以氯化亚锡为还原剂，对DEMDP进行了99Tcm标记，探索了最佳标记条件。
研究结果表明，标记反应的最佳条件为：8mgDEMDP，0． 4mg SnCl2·2H2O，反
应液pH值范围为1． 5-4． 8，反应温度为100℃，反应时间为30min，标记率可达95%
以上。标记物体外稳定性好，标记后室温下放置6h，标记率仍能保持在90%以上，
其稳定性满足动物实验要求。
观察了99Tcm-DEMDP在昆明小白鼠的体内分布情况，发现标记物的骨摄取较
高，且滞留时间长，但标记物的血清除较慢。同时动物实验结果表明标记物在肾
中分布高，提示标记物主要经过肾排泄。
研究了反相离子对HPLC分析99Tcm-DEMDP的条件。最佳淋洗条件为：
0． 1mol／L NaAC+0． 003mol／L TBA+2%EtAC+0． 0065mol／L DEMDP+0． 005mol／L
Sn2+，流速为2mL／min。得到了三种不同温度下DEMDP的99Tcm标记物的图谱，
并在相同淋洗条件下与99Tcm-MDP的图谱进行了比较，其结果表明DEMDP在加热
条件下进行99Tcm标记时不会发生分解生成MDP。
Diethyl Methylenebisphosphnate(DEMDP) is an analogue of Methylene Diphosphonate(MDP),which is currently in wide use as bone imaging agent in nuclear medicine. The synthesis of DEMDP and the characterization of metal complex have been studied in literature. But the labeling of DEMDP with 99Tcm and the biodistribution of the labeled compound have not been studied yet.
DEMDP has been characterized by IR,'H-NMR,31P-NMR and elemental analysis. Reduction of 99TcmO4- by stannous chloride in the presence of excess DEMDP leads to the product of 99Tcm-DEMDP complexes. The study indicates the best reaction conditions is:8mg DEMDP,0.4mg SnCl2PH:1.5-4.8,reacted temperature 100C,reacted time:30min.In this case,the labeling efficiency can go up to 95%.Furthermore 99Tcm-DEMDP can be kept stably at the room temperature in that the labeling percent is over 90% in six hours after being labeled. The biodistribution of 99Tcm-DEMDP in mice had been studied and It was found that the bone uptake of the labelld complex was quite high,but its blood clearance was slow.
For the characterization of the multi-component bone-scan agent 99Tcm-diphosphonate we have analysed the complex mixture with reversed phase ion pair chromatography (RP-IPC). The optimal condition of RP-IPC for 99Tcm -DEMDP is as follows:the eluent of 0.1mol/L + 0.003mol/LTBA +1-2%EtAC+0.0065mol/L DEMDP + 0.005mol/LSn2+;the flow rate of 2mL/min.With RP-IPC more than one peak were found within a separation-time of 40 min. When prepared under different temperature,it appears that the composition of the complexes is not the same.
基二膦酸(MDP)的类似物。文献中有关DEMDP的合成及其金属螯合物的研究已
有一些报道，但是，对于DEMDP的99Tcm标记及标记物的生物分布尚未见报道。
本文对由合成MDP的母液中提取出来的DEMDP进行了元素分析、核磁共
振、红外光谱、质谱分析，从而对其结构进行了表征。
以氯化亚锡为还原剂，对DEMDP进行了99Tcm标记，探索了最佳标记条件。
研究结果表明，标记反应的最佳条件为：8mgDEMDP，0． 4mg SnCl2·2H2O，反
应液pH值范围为1． 5-4． 8，反应温度为100℃，反应时间为30min，标记率可达95%
以上。标记物体外稳定性好，标记后室温下放置6h，标记率仍能保持在90%以上，
其稳定性满足动物实验要求。
观察了99Tcm-DEMDP在昆明小白鼠的体内分布情况，发现标记物的骨摄取较
高，且滞留时间长，但标记物的血清除较慢。同时动物实验结果表明标记物在肾
中分布高，提示标记物主要经过肾排泄。
研究了反相离子对HPLC分析99Tcm-DEMDP的条件。最佳淋洗条件为：
0． 1mol／L NaAC+0． 003mol／L TBA+2%EtAC+0． 0065mol／L DEMDP+0． 005mol／L
Sn2+，流速为2mL／min。得到了三种不同温度下DEMDP的99Tcm标记物的图谱，
并在相同淋洗条件下与99Tcm-MDP的图谱进行了比较，其结果表明DEMDP在加热
条件下进行99Tcm标记时不会发生分解生成MDP。
Diethyl Methylenebisphosphnate(DEMDP) is an analogue of Methylene Diphosphonate(MDP),which is currently in wide use as bone imaging agent in nuclear medicine. The synthesis of DEMDP and the characterization of metal complex have been studied in literature. But the labeling of DEMDP with 99Tcm and the biodistribution of the labeled compound have not been studied yet.
DEMDP has been characterized by IR,'H-NMR,31P-NMR and elemental analysis. Reduction of 99TcmO4- by stannous chloride in the presence of excess DEMDP leads to the product of 99Tcm-DEMDP complexes. The study indicates the best reaction conditions is:8mg DEMDP,0.4mg SnCl2PH:1.5-4.8,reacted temperature 100C,reacted time:30min.In this case,the labeling efficiency can go up to 95%.Furthermore 99Tcm-DEMDP can be kept stably at the room temperature in that the labeling percent is over 90% in six hours after being labeled. The biodistribution of 99Tcm-DEMDP in mice had been studied and It was found that the bone uptake of the labelld complex was quite high,but its blood clearance was slow.
For the characterization of the multi-component bone-scan agent 99Tcm-diphosphonate we have analysed the complex mixture with reversed phase ion pair chromatography (RP-IPC). The optimal condition of RP-IPC for 99Tcm -DEMDP is as follows:the eluent of 0.1mol/L + 0.003mol/LTBA +1-2%EtAC+0.0065mol/L DEMDP + 0.005mol/LSn2+;the flow rate of 2mL/min.With RP-IPC more than one peak were found within a separation-time of 40 min. When prepared under different temperature,it appears that the composition of the complexes is not the same.
引文
1) Buja L .M; Tofe A. J; et al ,sites and mechanisms of localization of technetium-99m phosphorus radiopharmaceuticals in acute myocardial infarcts and other tissues. J. Clin. Invest, 60:724-740; 1977．
2) Perez R, Cohen Y, Henry R,et al A new radiopharmaceutical for99mTc bone scanning. J .Nucl .Med .1972,13:788-789
3) Subramanian G, Mcafee J G, et al.Technetium-99mTc Methylene diphosphonate-A superior agent for skeletal imaging: Comparison with other technetium complexes. J .Nucl .Med .1975,16,744-755．
4) J .A .Sevan , A .J.Tofe et al .Tc-99m HMDP (Hydoxymethylene diphosphonate):A radiopharmaceutical for skeletal and acute myocardial infarct imaging J .Nucl. Med .21:961-966,1980．
5) Udalrich Buell, Edurad Kleinhans et al .A comparison of bone imaging with Tc-99m DPD and Tc-99m MDP. J .Nucl .MED. 23:214-217,1982．
6) 马寄晓 刘秀杰。实用临床核医学。原子能出版社，234-235，1990
7) 方瑞英，实用核医学显像技术，上海医科大学出版社。1996．
8) Huigen ,Y .M; Tji, T.G et alThe binding of 99mTc(Sn)-MDP complexes to human serum albumin and other blood-proteins determined with gel chromatography and ultrafiltration. Appl .Radiat .Isot .40:629-635,1989．
9) Savelkoul, T J .F; Van Ginkel ,J J. et al .A protein-binding and urinary excretion of 99mTc(Sn)-MDP and 99mTc-MDP. Int. J ,N ucl .Med .Biol.12:125-131,1985．
10) C L.DeLigny, W.J.Glsema et al.Bone seeking radiopharmaceuticals . Nucl .Med .Biol.17:161-179,1990．
11) Fogelman ,I;Citrin ,D .L et al . A clinical comparison of 99mTc-HEDP and 99mTc-MDP in the detection of bone metastases: concise communication.
J．Nucl．MED．20：98-101，1979．
12) 王吉欣，骨与骨髓诊断用放射性药物．见王吉欣，卢玉楷主编，放射性 药物学，原子能出版社，1999，258。
13) Francis M .D, Russell R .G .G and Fleisch H: Diphosphonates inhibit formation of Calcium phosphate crystals in vitro and pathological in vivo Science 165:1264-1266,1969
14) Fleisch H ,Russell R .G .G and Francis M .D: Diphosphonates inhibit hydroxyapatite dissolution in vitro and bone absoption in tissue culture and in vivo .Science 165:1262-1264
15) J Jowsey ,B.L .Riggs et al : The treatment of Osteoporosis wih disodium ethane-1-hydroxy-l,1-diphosphonate J .Lab .Clin .Med 77:574-585,1971
16) Jowsey J, Holley K .E et al: Effect of sodium etidronate in adult cats. J .Lab .Clin .Med .76:126-133,1970
17) R .Smith ,R .G .G .Russell et al : Diphosphonates and Pagets disease of bone .the Lancet .945-947,1971
18) 第三届全国核素治疗研讨会，李茂良。1999，16-30。
19) 李力力，缪增星，刘国正：二膦酸盐配合物治疗类风湿的有效性研究
20) Nylen. Studien uber organische phosphorverbindungen Diss uppsala,1930
21) Henryk Krawczyk. A convenient route for monodealkylation of diethyl phosphonates .Synthetic communication.27(18) ,3151-3161． (1997)
22) H.Gorican and D.Grdenic. Dialkyl dihydrogen methylenebisphosphonate and their metal salts. J.Chem.Soc.513(1964)
23) C.M.Mikulski,N.M.Karayannis et al . Dethyl methylene diphosphonate-diethyl hydrogen methylene diphosphonato-Vanadium() .Inorg .Nucl.Chem.L ETT. 8,225(1972)
24) C.M.Mikulski, N.M.Karayannis et al.Synthesis and characterization of diethylmethylene diposphonato metal complexes . J. Inorg.Nucl. Chem.1973 ,35 ,4011-4021 .
25) D.Grdenic and V.Jagodic. Separation of Germanium from Arsenic by solvent
extraction with dioctyl methylenebisphosphonic acid. J.Inorg.Nucl.Chem. 1964,16,167-170．
26) D.Grdenic and H.Gorican: Solvent extraction of Niobium and Tantalum. J .Inorg .Nucl .Chem .28:1451-1459,1956．
27) H.Gorican and D.Grdenic: Dialkyl dihydrogen methylenebisphosphonates as regents for solvent extraction of metals .Proc .Chem ,Soc 1960,288-289．
28) I.A .Potapova, G .A .Nevinsky et al Interaction of dNTP ,pyrophosphate and their analogs with the dNTP-binding sites of E.coli DNA polymerase Klenow fragment and human DNA polymerase a FEBS LETTERS .277:194-196,1990．
29) Nevinsky I.A, Potapova, G .A et al .Interaction of d dNTP-binding sites of human DNA polymerase a and klenow fragment of DNA polymerase from E.coli with nucleotide, pyrophosphate and their analogs.Mol.Biol 199024(1) ,104-116．
30) TjiJ .G; Gelsema, W J; Ligny de C .L . Determination of the mean oxidation state of Tc in 99Tc(Sn)EHDP,99mTc(Sn)EHDP, 99Tc(Sn)MDP and 99mTc(Sn)MDP complexes.Characterization of Tc(Ⅲ)-,Tc(Ⅳ)-and Tc(Ⅴ)-EHDP complexes .Appl .Radiat .Isot .(MS 443) . 1989．
31) Korteland ,J ; Dekker ,B .G; Ligny de C .L . The valence state of technetium-99 in its complexes with bleomycin ,l-hydroxy-ethylidene-1,1-diphosphonate and human serum albumin .Int. J .Appl .Radiat .Isot .31:315-318,1980．
32) Hambright P ;Mcrae J; Valk P .E .et al Chemistry of technetium radiopharmaceuticals .Exploration of the tissue distribution and oxidation state consequences of technetium(Ⅳ) in Tc-Sn-gluconate and Tc-Sn-EHDP using carrier 99Tc . J .Nucl .Med .16:478-482,1975．
33) J .A .G .M .Van Den Brand ,H .A.Das.B .G .Dekker and C . L .de Ligny. The influence of various 99mTc(Sn)EHDP complexes .Int J .Appl .Radiat .33:39-45． 1982
34) Shinzo Tanabe, Julius Zodda et al .Effect of pH on the formation of Tc(NaBH4) -MDP radiopharmaceutical analogues. Int J .Appl .Radiat .Isot . 34:1577-1584． 1983．
35) R J .A Nieuwland ,H .A .Das and C .L .De Ligny .Improvement of the reproducibility of ion-pair HPLC of 99mTc(Sn)EHDP complexes and the influence of the Sn(Ⅱ) concentration on the composition of the reaction mixture . Appl .Radiat .Isot .40:153-157． 1989．
36) Y .M .Huigen ,W J .Gelsema and C .L .De Ligny .Effect of the composition of the eluent in the chromatography of 99mTc-diphosphonate complexes. Appl .Radiat .Isot .39:381-384． 1988．
37) .G. J .De Groot, H. A. Das and C .L. De Ligny. The separation of 99mTc(Sn)EHDP complexes by HPLC and GPC . Appl. Radiat. Isot. 37,23-27,1986．
38) J.Kroesbergen ,W J. Gelsema and C.L.De Ligny.99mTc bone scanning agent-Preparation and gel chromatography of 99mTc(Sn)MDP complexes. Int. J.Nucl.Med.Biol. 12,419-424． 1986．
39) Shinzo Tanabe Julius P .Zodda et al. The biological distributions of some Technetium-MDP components isolated by anion exchange high performance liquid chromatograghy. J .Appl .Radiat .Isot.34,1585-1592,1983．
40) Ase Handeland ,Mette .Winderen Lindegard and Dag-Erik .Heggli. Biodistribution of anionic separated MDP complexes from different MDP preparations .Eur J .Nucl .Med .(1989) 15:609-611
41) 5． J .P .Zodda, Shinzo Tanabe et al. The effect of formulation conditions on the distribution of 99mTc(NaBH4) -HEDP components separated by anion exchange HPLC. Appl .Radiat .Isot .37,345-354,1986．
42) J.A.G.M.Van Den Brand ,H.A Das ,et al The Influence of Experimental Conditions on he Formation of Various 99mTc (Sn)-HEDP Complexes. Int. J. Appl. Radiat. Isot.32,637(1981)
43) Kroesbergen J,Gelsema W.J and de Ligny C. L. 99mTc bone scanning agents-Ⅲ.Preparation and gel chromatography of 99mTc(Sn)MDP complexes. Int.Nucl. MedBiol. 12,419(1986)
44) Y.M.Huigen, T.G.Tji et al Anion exchange chromatography 99mTc (Sn)-MDPcomplexes: Influence of eluent composition determination of void
volume and change ofthe main component．Appl．Ridiat．Isot．39，25-30，1988．
45) G．J．De Groot，H.A．Das and C．L．De Ligny．The separation of 99mTc(Sn)HEDP complexes by HPLC and GEL．Appl．Radiat．Isot．37，23-27． 1986．
46) R．J．A．Nieuwland,H．A．Das and C．L．De．Ligny．Improvement of the reproducibility of ion-pair HPLC of 99mTc(Sn)EHDP complexes and the influence of the Sn(Ⅱ)concentration on the composition of the reaction mixture．Appl． Radiat．Isot．40，153-157． 1989．
47) Y.M．Huigen，W．J．Gelseman and C．L．De Ligny．Effect of the composition of the eluent in the chromatography of 99mTc-diphosphonate complexes．Appl．Radiat． Isot．39，381-384． 1988．
48) 周申范 宋敬埔 王乃岩。色谱理论及应用。北京理工大学出版社，273-275 (1994)
49) Srivastava S．C，Meinken G．E，Richards L．A．et al(1982) Proc．3rd World Congress of Nucl．Med and Biol，Paris．V2，631：Pergamon Press Oxford．
2) Perez R, Cohen Y, Henry R,et al A new radiopharmaceutical for99mTc bone scanning. J .Nucl .Med .1972,13:788-789
3) Subramanian G, Mcafee J G, et al.Technetium-99mTc Methylene diphosphonate-A superior agent for skeletal imaging: Comparison with other technetium complexes. J .Nucl .Med .1975,16,744-755．
4) J .A .Sevan , A .J.Tofe et al .Tc-99m HMDP (Hydoxymethylene diphosphonate):A radiopharmaceutical for skeletal and acute myocardial infarct imaging J .Nucl. Med .21:961-966,1980．
5) Udalrich Buell, Edurad Kleinhans et al .A comparison of bone imaging with Tc-99m DPD and Tc-99m MDP. J .Nucl .MED. 23:214-217,1982．
6) 马寄晓 刘秀杰。实用临床核医学。原子能出版社，234-235，1990
7) 方瑞英，实用核医学显像技术，上海医科大学出版社。1996．
8) Huigen ,Y .M; Tji, T.G et alThe binding of 99mTc(Sn)-MDP complexes to human serum albumin and other blood-proteins determined with gel chromatography and ultrafiltration. Appl .Radiat .Isot .40:629-635,1989．
9) Savelkoul, T J .F; Van Ginkel ,J J. et al .A protein-binding and urinary excretion of 99mTc(Sn)-MDP and 99mTc-MDP. Int. J ,N ucl .Med .Biol.12:125-131,1985．
10) C L.DeLigny, W.J.Glsema et al.Bone seeking radiopharmaceuticals . Nucl .Med .Biol.17:161-179,1990．
11) Fogelman ,I;Citrin ,D .L et al . A clinical comparison of 99mTc-HEDP and 99mTc-MDP in the detection of bone metastases: concise communication.
J．Nucl．MED．20：98-101，1979．
12) 王吉欣，骨与骨髓诊断用放射性药物．见王吉欣，卢玉楷主编，放射性 药物学，原子能出版社，1999，258。
13) Francis M .D, Russell R .G .G and Fleisch H: Diphosphonates inhibit formation of Calcium phosphate crystals in vitro and pathological in vivo Science 165:1264-1266,1969
14) Fleisch H ,Russell R .G .G and Francis M .D: Diphosphonates inhibit hydroxyapatite dissolution in vitro and bone absoption in tissue culture and in vivo .Science 165:1262-1264
15) J Jowsey ,B.L .Riggs et al : The treatment of Osteoporosis wih disodium ethane-1-hydroxy-l,1-diphosphonate J .Lab .Clin .Med 77:574-585,1971
16) Jowsey J, Holley K .E et al: Effect of sodium etidronate in adult cats. J .Lab .Clin .Med .76:126-133,1970
17) R .Smith ,R .G .G .Russell et al : Diphosphonates and Pagets disease of bone .the Lancet .945-947,1971
18) 第三届全国核素治疗研讨会，李茂良。1999，16-30。
19) 李力力，缪增星，刘国正：二膦酸盐配合物治疗类风湿的有效性研究
20) Nylen. Studien uber organische phosphorverbindungen Diss uppsala,1930
21) Henryk Krawczyk. A convenient route for monodealkylation of diethyl phosphonates .Synthetic communication.27(18) ,3151-3161． (1997)
22) H.Gorican and D.Grdenic. Dialkyl dihydrogen methylenebisphosphonate and their metal salts. J.Chem.Soc.513(1964)
23) C.M.Mikulski,N.M.Karayannis et al . Dethyl methylene diphosphonate-diethyl hydrogen methylene diphosphonato-Vanadium() .Inorg .Nucl.Chem.L ETT. 8,225(1972)
24) C.M.Mikulski, N.M.Karayannis et al.Synthesis and characterization of diethylmethylene diposphonato metal complexes . J. Inorg.Nucl. Chem.1973 ,35 ,4011-4021 .
25) D.Grdenic and V.Jagodic. Separation of Germanium from Arsenic by solvent
extraction with dioctyl methylenebisphosphonic acid. J.Inorg.Nucl.Chem. 1964,16,167-170．
26) D.Grdenic and H.Gorican: Solvent extraction of Niobium and Tantalum. J .Inorg .Nucl .Chem .28:1451-1459,1956．
27) H.Gorican and D.Grdenic: Dialkyl dihydrogen methylenebisphosphonates as regents for solvent extraction of metals .Proc .Chem ,Soc 1960,288-289．
28) I.A .Potapova, G .A .Nevinsky et al Interaction of dNTP ,pyrophosphate and their analogs with the dNTP-binding sites of E.coli DNA polymerase Klenow fragment and human DNA polymerase a FEBS LETTERS .277:194-196,1990．
29) Nevinsky I.A, Potapova, G .A et al .Interaction of d dNTP-binding sites of human DNA polymerase a and klenow fragment of DNA polymerase from E.coli with nucleotide, pyrophosphate and their analogs.Mol.Biol 199024(1) ,104-116．
30) TjiJ .G; Gelsema, W J; Ligny de C .L . Determination of the mean oxidation state of Tc in 99Tc(Sn)EHDP,99mTc(Sn)EHDP, 99Tc(Sn)MDP and 99mTc(Sn)MDP complexes.Characterization of Tc(Ⅲ)-,Tc(Ⅳ)-and Tc(Ⅴ)-EHDP complexes .Appl .Radiat .Isot .(MS 443) . 1989．
31) Korteland ,J ; Dekker ,B .G; Ligny de C .L . The valence state of technetium-99 in its complexes with bleomycin ,l-hydroxy-ethylidene-1,1-diphosphonate and human serum albumin .Int. J .Appl .Radiat .Isot .31:315-318,1980．
32) Hambright P ;Mcrae J; Valk P .E .et al Chemistry of technetium radiopharmaceuticals .Exploration of the tissue distribution and oxidation state consequences of technetium(Ⅳ) in Tc-Sn-gluconate and Tc-Sn-EHDP using carrier 99Tc . J .Nucl .Med .16:478-482,1975．
33) J .A .G .M .Van Den Brand ,H .A.Das.B .G .Dekker and C . L .de Ligny. The influence of various 99mTc(Sn)EHDP complexes .Int J .Appl .Radiat .33:39-45． 1982
34) Shinzo Tanabe, Julius Zodda et al .Effect of pH on the formation of Tc(NaBH4) -MDP radiopharmaceutical analogues. Int J .Appl .Radiat .Isot . 34:1577-1584． 1983．
35) R J .A Nieuwland ,H .A .Das and C .L .De Ligny .Improvement of the reproducibility of ion-pair HPLC of 99mTc(Sn)EHDP complexes and the influence of the Sn(Ⅱ) concentration on the composition of the reaction mixture . Appl .Radiat .Isot .40:153-157． 1989．
36) Y .M .Huigen ,W J .Gelsema and C .L .De Ligny .Effect of the composition of the eluent in the chromatography of 99mTc-diphosphonate complexes. Appl .Radiat .Isot .39:381-384． 1988．
37) .G. J .De Groot, H. A. Das and C .L. De Ligny. The separation of 99mTc(Sn)EHDP complexes by HPLC and GPC . Appl. Radiat. Isot. 37,23-27,1986．
38) J.Kroesbergen ,W J. Gelsema and C.L.De Ligny.99mTc bone scanning agent-Preparation and gel chromatography of 99mTc(Sn)MDP complexes. Int. J.Nucl.Med.Biol. 12,419-424． 1986．
39) Shinzo Tanabe Julius P .Zodda et al. The biological distributions of some Technetium-MDP components isolated by anion exchange high performance liquid chromatograghy. J .Appl .Radiat .Isot.34,1585-1592,1983．
40) Ase Handeland ,Mette .Winderen Lindegard and Dag-Erik .Heggli. Biodistribution of anionic separated MDP complexes from different MDP preparations .Eur J .Nucl .Med .(1989) 15:609-611
41) 5． J .P .Zodda, Shinzo Tanabe et al. The effect of formulation conditions on the distribution of 99mTc(NaBH4) -HEDP components separated by anion exchange HPLC. Appl .Radiat .Isot .37,345-354,1986．
42) J.A.G.M.Van Den Brand ,H.A Das ,et al The Influence of Experimental Conditions on he Formation of Various 99mTc (Sn)-HEDP Complexes. Int. J. Appl. Radiat. Isot.32,637(1981)
43) Kroesbergen J,Gelsema W.J and de Ligny C. L. 99mTc bone scanning agents-Ⅲ.Preparation and gel chromatography of 99mTc(Sn)MDP complexes. Int.Nucl. MedBiol. 12,419(1986)
44) Y.M.Huigen, T.G.Tji et al Anion exchange chromatography 99mTc (Sn)-MDPcomplexes: Influence of eluent composition determination of void
volume and change ofthe main component．Appl．Ridiat．Isot．39，25-30，1988．
45) G．J．De Groot，H.A．Das and C．L．De Ligny．The separation of 99mTc(Sn)HEDP complexes by HPLC and GEL．Appl．Radiat．Isot．37，23-27． 1986．
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