| 1. |
Tetsuya Komene, Kotoko Kai, Kiyoko Kinpara, Tomoaki Sato, Eisaku Hokazono, Tatsuo Shimosawa, Susumu Osawa, Masanori Seimiya, Development of a plasma maltose assay method as a screening test for upper gastrointestinal disorders, Annals of Clinical Biochemistry, 10.1177/00045632231224218, 2024.01, Background and objective: The disaccharide loading test is a method to assess gastric mucosal damage. Since Trelan-G75, which is used for the sugar tolerance test, contains disaccharide maltose, if maltose is detected at a high sensitivity in the sample blood used in the sugar tolerance test, screening for upper gastrointestinal mucosal damage can be made simultaneously with the sugar tolerance test for the diagnosis of diabetes.
Methods: Glucose-6-phosphate is generated by treating maltose with maltose phosphorylase, β-phosphoglucomutase, and glucose-1,6-bisphosphate. Then, change in the absorbance at 405 nm is measured by the enzymatic cycling method using Thio-NADP, β-NADPH, and Glucose-6-phosphate dehydrogenase. After evaluating the optimal condition for this method, it is mounted on an automatic biochemical analyzer, and samples after the sugar tolerance test were assayed.
Results: Regarding the performance of this method, the repeatability was 10-50 μmol/L with a CV of ≤1.1%. Concerning the assay range, a curve passing the origin with a range of linearity up to 120 μmol/L was obtained. No effect of dyes or sugars in the blood was noted. As a result of application to patients with gastric mucosal disorders (those who had a health checkup), significant differences were observed depending on the stage of atrophic gastritis.
Discussion: This method has a high sensitivity and a high precision and can be used for high-speed analysis on an automatic analyzer. It has the potential to be used as a screening test for gastric mucosal damage.. |
| 2. |
Eisaku Hokazono, Saori Fukumoto, Takeshi Uchiumi, SusumuOsawa, Pyrophosphate detection method using 5-Br-PAPS to detect nucleic acid amplification - Application to LAMP method -, Analytical Biochemistry, https://doi.org/10.1016/j.ab.2023.115371, 684, https://doi.org/10.1016/j.ab.2023.115371, 2024.01, Genetic testing has been increasingly used in several fields. In many applications, nucleic acid amplification technology is required. However, current methods to detect nucleic acid amplification require expensive reagents and special equipment or exhibit limited sensitivity, which hinders their use. To address this issue, this study reports an assay method for detecting occurrence of acid amplification in post-amplification samples using pyrophosphate, a highly sensitive byproduct of nucleic acid amplification. The method proposed requires two reagents and an automated analyzer. First, hydrogen peroxide is derived from pyrophosphate, an indicator of nucleic acid amplification, and the oxidizing power of hydrogen peroxide is used to produce Fe (III) from Fe (II). The specific metal chelator 5-Br-PAPS forms a complex with the trivalent iron produced, resulting in a highly sensitive coloration. The within-run reproducibility of our method (n = 20) was less than 3.67% at each concentration tested, and the detection limit was 0.075 μmol/L, sufficient for quantitative analysis. The technique described could detect pyrophosphate in a sample that was amplified using the loop-mediated isothermal amplification method after only 10 min. Therefore, the proposed method has the potential to be a new, rapid, and simple detection technique for amplified nucleic acids.. |
| 3. |
河野 弥季、石田 綾乃、外園 栄作、大澤 進, 赤血球中のphospholipase-D活性測定法の構築, 医学検査, 10.14932/jamt.20-86, 70, 2, 198-204, Vol.70, No2, 198-204, 2021.04. |
| 4. |
Eisaku Hokazono, Eri Ota, Taiki Goto, Saori Fukumoto, YuzoKayamori, Takeshi Uchiumi, SusumuOsawa, Development of a protein assay with copper chelator chromeazurol B, based on the biuret reaction, Analytical Biochemistry, https://doi.org/10.1016/j.ab.2021.114320, https://doi.org/10.1016/j.ab.2021.114320, 2021.07, This study aimed to provide a novel and highly sensitive protein assay based on the biuret reaction and using chromeazurol B, a metal chelate compound. The method consists of two reagents and an automated analyzer. First, a complex of copper and protein (biuret reaction) is formed. Second, a chelating reagent containing chromeazurol B forms a three-dimensional complex of protein, copper, and chromeazurol B at neutral pH, resulting in highly sensitive coloration. The intra-assay (n = 20) variation for the three levels was 3.54 % or lower at each concentration. Each response with α, β-, and γ-globulin was 103.8 % and 104.3 %, respectively, against albumin. The molar absorption coefficient (ε) of the present method was 2.5 × 105 m2/mol against human albumin, higher than that of the commercially available Lowry method (ε = 8.7 × 104 m2/mol), which is based on the same principle. The correlation test for the pyrogallol method with 30 urine samples showed good performance (r = 0.961). The method described here (the Biuret-based CAB method) is a more sensitive and rapid assay than the Lowry method, and it may also be applied to biological samples because of its similar reactivity towards various proteins.. |
| 5. |
Sonoko Yoshihiro, Takuya Ishigaki, Hayato Ookurano, Fumi Yoshitomi, Taeko Hotta, Dongchon Kang, Eisaku Hokazono and Yuzo Kayamori, New colorimetric method with bromocresol purple for estimating the redox state of human serum albumin, J. Clin. Biochem. Nutr., https://doi.org/10.3164/jcbn.20-10, 67, 3, 257-262, Volume 67 Issue 3 Pages 257-262, 2020.03, Oxidative damage results in protein modification and is observed in many diseases, such as heart failure and renal insufficiency. Human serum albumin is an index of oxidative change and is conventionally measured using high-performance liquid chromatography (HPLC). Although this method is more sensitive than the colorimetric method, it is time-consuming for clinical practice and the sera must be stored at –80°C before analysis. To overcome these limitations, in the present study we developed a new reagent for a more rapid and convenient quantification of oxidative stress, involving determination of the ratio of human nonmercaptalbumin to total albumin using a colorimetric method with bromocresol purple. The clinical utility of the developed reagent was confirmed by demonstrating the consistently higher oxidative stress levels in dialysis patients than in healthy control subjects, matching the results of the conventional HPLC method. This novel approach could be a valuable tool for immediate estimation of the state of oxidative stress during the course of disease and treatment, and could aid clinical treatment decisions.. |
| 6. |
佐藤 有華, 清宮 正徳, 吉田 俊彦, 澤部 祐司, 外園 栄作, 大澤 進, 松下 一之, 生化学自動分析装置を用いたインドシアニングリーン測定法の検討, 生物試料分析科学会 生物試料分析, 42, 4, 200-204, Vol.42, No4, 200-204, 2019.09. |
| 7. |
外園 栄作、保坂 洸喜、秋本 卓、川満 紀子、 酒本 美由紀、堀田 多恵子、康 東天、栢森 裕三, 96穴プレートを用いた簡便な尿中Tamm-Horsfall Protein(THP）の測定法の検討, 生物試料分析科学会 生物試料分析, 42, 5, 248-255, Vol.42, No5, 248-255, 2019.12, Tamm-Horsfall protein (THP) is primarily synthesized in the thick ascending limb of the loop of Henle and is the most abundant protein in human urine. THP is excreted at an average rate of 50–100 mg/day. Deviations from this level of urinary excretion is an indicator of various clinical conditions and diseases. THP is generally measured by an ELISA-based method, which suffers from limitations including inaccuracy due to protein aggregation and high reagent costs. Herein, a simple method to measure THP using cost-effective reagents and a 96-well micro-plate was developed. The average within-run CVs were 4.5–9.6% (n = 10). The correlation between the values obtained using the newly developed method (y) and conventional HPLC methods (x) was: y = 1.057x -5.702 (r = 0.978, n = 35). Thus, this novel method will be useful for further studies to clarify the clinical significance of THP.. |
| 8. |
栢森 雄三、外園 栄作, 臨床化学領域における生物試料分析, 福岡医学雑誌, 110, 3, 133-145, 2019.09. |
| 9. |
Miki Kawano, Eisaku Hokazono, Susumu Osawa, Shouichi Sato, Takiko Tateishi, Masahiro Manabe, Hirotaka Matsui, Yuzo Kayamori, A novel assay for triglycerides using glycerol dehydrogenase and a water-soluble formazan dye, WST-8, Annals of Clinical Biochemistry, 10.1177/0004563219830715, 56, 4, 442-449, Vol.56(4), 442-449,2019, 2019.01, Background
The GPO-POD chromogenic method is well known that peroxidase is affected by reducing agents, and recently, it has been reported that some materials affect its activity. Moreover, there is a high possibility of non-specific reaction as the method uses many enzymes. Against this background, we developed a simpler assay method for TGs without using peroxidase.
Methods
TGs were hydrolysed to glycerol and fatty acids by LPL followed by the reduction of the glycerol dehydrogenase (GDH) to dihydroxyacetone with simultaneous production of NADH from NAD+. To overcome incomplete conversion of glycerol to dihydroxyacetone by GDH at equilibrium, we added 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt (WST-8) to the reaction mixture to remove NADH, allowing the reaction to complete while showing stoichiometric production of reduced WST-8.
Results
The reaction was linear up to 6.4 mmol/L. The mean intra-assay (n = 20) and inter-assay (n = 20) imprecision, as determined by replicate analysis of three pooled human serum samples with different TG concentrations, were 1.1-2.3% and 1.1-1.5% coefficient of variation (%CV), respectively. No interference by 2.5 g/L haemoglobin, 65 μmol/L free bilirubin, and 359 μmol/L conjugated bilirubin were observed. The equation obtained in comparison with that by the GPO-POD method including endogenous glycerol eliminating step was: y = 1.0002x + 0.0395 mmol/L; r = 0.999; Sy/x = 0.049 mmol/L; n = 97.
Conclusion
Our method is an accurate, yet simpler and more sensitive, alternative method for the quantitative analysis of TGs.. |
| 10. |
Yuka Sato,Masanori Seimiya,Toshihiko Yoshida,Yuji Sawabe, Eisaku Hokazono, Susumu Osawa, Kazuyuki Matsushita, Development of a simple indocyanine green measurement method using an automated biochemical analyser, Annals of Clinical Biochemistry, 10.1177/0004563217745895, 55, 4, 491-495, 2018.07, Background: The indocyanine green retention rate is important for assessing the severity of liver disorders. In the
conventional method, blood needs to be collected twice. In the present study, we developed an automated indocyanine
green method that does not require blood sampling before intravenous indocyanine green injections and is applicable to
an automated biochemical analyser.
Methods: The serum samples of 471 patients collected before and after intravenous indocyanine green injections and
submitted to the clinical laboratory of our hospital were used as samples. The standard procedure established by the
Japan Society of Hepatology was used as the standard method. In the automated indocyanine green method, serum
collected after an intravenous indocyanine green injection was mixed with the saline reagent containing a surfactant,
and the indocyanine green concentration was measured at a dominant wavelength of 805 nm and a complementary
wavelength of 884 nm.
Results: The coefficient of variations of the within- and between-run reproducibilities of this method were 2% or lower,
and dilution linearity passing the origin was noted up to 10 mg/L indocyanine green. The reagent was stable for four
weeks or longer. Haemoglobin, bilirubin and chyle had no impact on the results obtained. The correlation coefficient
between the standard method (x) and this method (y) was r¼0.995; however, slight divergence was noted in turbid
samples.
Conclusion: Divergence in turbid samples may have corresponded to false negativity with the standard procedure.
Our method may be highly practical because blood sampling before indocyanine green loading is unnecessary and
measurements are simple.. |
| 11. |
Shou Terada,Miyuki Sakemoto,Yukiko Kawanobe,Miki Kawano,Takiko Tateishi,Taeko Hotta,Dongchon Kang,Yuzo Kayamori and Eisaku Hokazono, Establishment of a rapid and simple assay for measuring serum trehalase activity, Int J Anal Bio-Sci, 6, 2, 19-24, Vol. 6, No2 (2018), 2018.06, Trehalase (TREH, trehalose-1-glucohydrolase, EC 3.2.1.28) catalyzes the hydrolysis of α,α-trehalose (1-α-D-glucopyranosyl-α-D-glucopyranoside) to form two molecules of glucose. TREH has been identified in the brush border membranes of the kidney, liver, and small intestine of mammals, but its physiological role has not yet been clarified. Some methods for measuring TREH activity have been reported, but they have been time-intensive, and its suitability for quantitative analysis could not be ascertained. Therefore, we developed a simple and rapid enzymatic assay for measuring serum TREH activity using a Hitachi 7600 type automated analyzer and evaluated the assay performance. The precision, linearity, detection limit, and recovery test were good. This new method effectively measures the serum TREH activity and can be easily integrated with an automated analyzer for rapid, high performance assays. This method can also support further research on understanding TREH activity.. |
| 12. |
Kenji Konishi, Shigeru Ueda, Miki Kawano, Susumu Osawa, Tomohiro Tamura, Eisaku Hokazono, Yuzo Kayamori, Shin-ich Sakasegawa, Characterization and application of a novel nicotinamide mononucleotide adenylyltransferase from Thermus thermophilus HB8, Journal of Bioscience and Bioengineering, Japan, 125, 4, 385-389, Volume 125(4), 385-389, 2018.04, Herein, we describe a novel enzymatic cycling method to measure nicotinamide mononucleotide (NMN) or nicotinic
acid mononucleotide (NaMN), which are precursors of NAD biosynthesis. A gene encoding a NMN adenylyltransferase
(NMNAT, EC 2.7.7.1) homologue was identified in Thermus thermophilus HB8. The gene from T. thermophilus
(TtNMNAT) was engineered for expression in Escherichia coli and the recombinant enzyme found to be stable,
retaining full activity after incubation for 45 min at 70°C. The Km values for NMN and ATP were calculated to be 0.263
and 1.27 mM, respectively, with a Vmax value of 60.3 μmol/min/mg. TtNMNAT was successfully applied to the
colorimetric NMN or NaMN assays, which employed (i) adenylation of NMN to NAD by TtNMNAT or adenylation of
NaMN to deamido-NAD (NaAD) by TtNMNAT followed by amidation of NaAD to NAD by NAD synthetase (NADS, EC
6.3.1.5) and (ii) a NAD cycling reaction using 12α-hydroxysteroid dehydrogenase (12α-HSD, EC 1.1.1.176) and
diaphorase (DI, EC 1.6.99.3) to accumulate reduced WST-8. This enzymatic cycling method enabled detection of 0.5
μM (12.2 nM in the reaction mixture) NMN or NaMN in an automatic clinical analyzer.. |
| 13. |
Takiko Tateishi, Yuki Matsuba, Kasumi Shimokawa, Maiko Kuwahara, Miki Kawano, Yuki Tanaka, Masahiro Manabe, Masanori Okuma, Katsuyoshi Ikeda, Hirotaka Matsui, Eisaku Hokazono, Yuzo Kayamori, Preliminary study of a high-sensitivity method to determine sarcosine in urine high-performance liquid chromatography, Int J Anal Bio-Sci, 5, 3, 43-51, Vol. 5, No 3(2017), 2017.05. |
| 14. |
Eri Ohta, Eisaku Hokazono, Takiko Tateishi, Miki Kawano, Yuzo Kayamori, Development of an enzymatic assay for ethanolamine in plasma, Int J Anal Bio-Sci, 4, 4, 110-116, Vol. 4, No 4 (2016), 2016.12, Ethanolamine (Etn) contributes to the generation of phosphatidyl ethanolamine (PE) in vivo. However, its plasma concentration and clinical significance in humans are still unclear. Therefore, we developed a simple, rapid enzymatic method using an amine oxidase and copper from Arthrobacter sp. (AAO) (EC 1.4.3.6). We found a strong correlation between the values obtained with this method and those obtained with the HPLC (high performance liquid chromatography)-based method (r = 0.89). Thus, it is suitable for routine clinical use in the labora- tory. Moreover, we would like to examine the clinical significance of Etn.. |
| 15. |
垰田 直美, 外園 栄作, 篠原 克幸, 大澤 進, 過ヨウ素酸ナトリウムを用いた血中ICG検査法の自動分析への試み
, 日本臨床自動化学会誌 , 41, 3, 270-277, 2016.Vol 41-No.3，270-277, 2016.06, The IV infusion of indocyanine green (ICG) is done during liver and circulatory function examinations. The standard procedure is a manual operation using a spectrophotometer, sometimes requiring drawing of blood before and after the medication. Therefore, the results include two kinds of errors: the first is from the differences in skill between operators, and the second is from the properties of the serum sample, such as hemolytic hemoglobin and chyle.
We aimed to resolve those problems. In this study, it did not need to be drawn before administration. Oxygenation of ICG with sodium periodate was used. The color of ICG at 805 nm disappears when sodium periodate is added. We applied this method to an automated biochemical analyzer, Hitachi7170.
The average within-run CVs were 0.97 – 3.13% (n=10) at 0.1 – 1.0 mg/dL. When we used 55 serum samples with added ICG, the correlation between the values obtained with the present method (y) and the manual method (x) was: y = 1.04x (r=0.999, Syx = 0.01).
The present method shows good performance evaluation and can be easily applied to automated analyzers.. |
| 16. |
Eri Ota, Shin-ich Sakasegawa, Shigeru Ueda, Kenji Konishi, Masaru Akimoto, Takiko Tateish, Eisaku Hokazono, Yuzo Kayamori, Preliminary evaluation of an improved enzymatic assay method for measuring potassium concentrations in serum, Clinica Chimica Acta, 73-75, Volume 446, 2015.06, Abstract
Background
K+ has important physiological functions. K+ concentrations in serum are generally determined using ion-selective electrodes (ISEs), though measurement using reagents in aqueous medium is also useful.
Methods
K+ concentrations were measured using recombinant inosine 5′-monophosphate dehydrogenase (IMPDH), which was activated only by K+ and NH4+. Exogenous NH4+ and endogenous NH4+ were eliminated using glutamine synthase.
Results
Regression analysis of the enzymatic assay (y) vs. the ISE method (x) gave the following relation: y = 1.03x + 0.09 (n = 54, Sy,x = 0.06 mmol/l). The linear range was up to 12 mmol/l when 1 U/ml IMPDH was used.
Conclusion
Advantages of the proposed assay method are: (i) the measured range is wider than that of existing enzymatic methods; (ii) the conditions for K+ determination can be maintained constant, regardless of the amount of NH4+ in the analyte and reagents; and (iii) the elimination system is simpler because the recombinant IMPDH is stimulated by only K+ and NH4+ and is unaffected by biological materials.. |
| 17. |
Masaru Akimoto, Eisaku Hokazono, Eri Ota, Takiko Tateish, Yuzo Kayamori, Highly sensitive reversed-phase high-performance liquid chromatography assay for the detection of Tamm–Horsfall protein in human urine, Annals of Clinical Biochemistry, 75-84, Volume 53, 2015.04, Abstract
Background: Tamm–Horsfall protein (also known as uromodulin) is the most abundant urinary protein in healthy individuals. Since initially characterized by Tamm and Horsfall, the amount of urinary excretion and structural mutations of Tamm–Horsfall protein is associated with kidney diseases. However, currently available assays for Tamm–Horsfall protein, which are mainly enzyme-linked immunosorbent assay-based, suffer from poor reproducibility and might give false negative results.
Methods: We developed a novel, quantitative assay for Tamm–Horsfall protein using reversed-phase high-performance liquid chromatography. A precipitation pretreatment avoided urine matrix interference and excessive sample dilution. High-performance liquid chromatography optimization based on polarity allowed excellent separation of Tamm–Horsfall protein from other major urine components.
Results: Our method exhibited high precision (based on the relative standard deviations of intraday [42.77%] and interday [45.35%] repetitions). The Tamm–Horsfall protein recovery rate was 100.0–104.2%. The mean Tamm–Horsfall protein concentration in 25 healthy individuals was 31.618.8 mg/g creatinine. There was a strong correlation between data obtained by high-performance liquid chromatography and enzyme-linked immunosorbent assay (r¼0.906), but
enzyme-linked immunosorbent assay values tended to be lower than high-performance liquid chromatography values at low Tamm–Horsfall protein concentrations.
Conclusions: The high sensitivity and reproducibility of our Tamm–Horsfall protein assay will reduce the number of false negative results of the sample compared with enzyme-linked immunosorbent assay. Moreover, our method is superior to other high performance liquid chromatography methods, and a simple protocol will facilitate further research on the physiological role of Tamm–Horsfall protein.. |
| 18. |
Akiyo Yumura, Eisaku Hokazono, Katsuyoshi Ikeda, Konen Obayashi, Yukio Ando, Susumu Osawa, Development of the enzymatic assay for whole blood choline using an automated biochemical analyzer, Int J Anal Bio-Sci, 2, 1, 23-32, Vol. 2, No 1 23-32, 2014.01. |
| 19. |
大澤 進, 石橋郁佳, 木内幸子, 外園 栄作, 栢森 裕三, 糖負荷試験の検体を利用した胃粘膜検査法の開発, 日本臨床自動化学会誌 , 38, 3, 270-277, 2013.Vol 38-No.3，270-277, 2013.03, The examination of gastric mucosa is performed both test gastrointestinal contrasting with X rays and pepsinogen test in serum. However, the being bombed with X-rays and an autoanalysis apparatus for exclusive immunoassay are necessary to take these tests.
We developed instead of maltose permeability test of gastric mucosa using the oral glucose tolerance samples. Enzymatic method for oligosaccharide is changed from endogenous glucose to glucose-6-phosphate by hexokinase and was eliminated. The oligosaccharide which was absorbed from a gastric mucosa is determined of the α-glucosidase-glucoseoxidase-peroxidase detection system as the determination of equivalent to oligosaccharide concentration. This method could discover a gastric mucosa disorder easily.
. |
| 20. |
Shinya Sugimoto, Masaru Akimoto, Akira Hayakawa, Eisaku Hokazono, Susumu osawa, Development of an assay of seven biochemical items, HbA1c, and hematocrit using a small amount of blood collected from the fingertip, Clinica Chimica Acta, 414, 192-197, 2012.01. |
| 21. |
Eisaku Hokazono , Hideto Tamezane, Taeko Hotta, Yuzo Kayamori, Susumu Osawa, Enzymatic Assay of Phosphatidylethanolamine in Serum using Amine Oxidase from Arthrobacter sp., Clinica Chimica Acta, 10.1016/j.cca.2011.04.023, Vol.412, 1436-1440, 2011.07. |
| 22. |
外園 栄作，吉田 彩香，湯村 旭代、大澤 進, 虚血性心疾患の新たなマーカーとしての血漿遊離コリンの酵素的測定法の開発, 医学検査, 59, 12, 1281-1286, 2010.Vol.59 No.12 p1281-1286, 2010.12, っゆ. |
| 23. |
外園 栄作，小島 夫美子，大澤 進, キサンテン系色素を用いた新しい尿沈渣染色液・保存液の検討− 第1報 染色色素の選定を中心に −, 医学検査, 59, 10, 1158-1164, 2010.Vol.59 No.10 p1158-1164, 2010.10. |
| 24. |
外園 栄作, 甲木 宏美, 小野 美由紀, 堀田 多恵子, 栢森 裕三, 大澤 進, 「マイクロTP-AR(2)」による尿中総蛋白測定試薬の基礎的性能評価, 日本臨床自動化学会誌 , 35, 5, 920-926, 2010.Vol 35-No.5，920-926, 2010.11. |
| 25. |
外園 栄作, 中野 倫太, 小口 雄二, 大澤 進, MRI造影剤の影響を回避した新しい血清カルシウム測定試薬の開発, 生物試料分析科学会 生物試料分析, 33, 2, 191-197, Vol.33, No2, 191-197, 2010.03, We previously reported a serum calcium measurement method using Chlorophosphonazo- III (CPZ-III). In this method, the reagent stability is higher than in conventional methods to react samples in an acid medium. High specificity to calcium is also achieved due to the addition of a surfac- tant to the reagent. However, gadolinium, which is used as a contrast agent for magnetic resonance imaging (MRI), causes a positive error in the measurement. Therefore, we have improved our previous method, and developed a new measurement reagent that is not affected by gadolinium. As a result, it was confirmed not to be unaffected by gadolinium to 2.0 mmol/L. Finally, even if our new method is employed just after using a contrast agent, the serum calcium tests should be unaffected. We expect this method to be used as a routine and emergency test in clinical laboratories because it is stable, accurate, and does not contain any heavy metals, dangerous materials, or mutagenic substances.. |
| 26. |
外園 栄作，上野 智美，小野 美由紀，堀田 多恵子，栢森 裕三，大澤 進, 小型自動分析装置日立クリニカルアナライザーS40への新しい血清カルシウム測定試薬の応用検討, 日本臨床自動化学会誌 , 34, 5, 583-859, 2009.Vol 34-5,853-859, 2009.10. |
| 27. |
Eisaku Hokazono, Susumu Osawa, Tomota Nakano, Yukari Kawamoto, Yuji Oguchi, Taeko Hotta, Yuzo Kayamori, Dongchon Kang, Yuichiro Cho, Kiyoko Shiba, Kenji Sato, Development of a new measurement method for serum calcium with chlorophosphonazo-III, Annals of Clinical Biochemistry, 10.1258/acb.2009.008099, 46, 296-301, 46: 296–301, 2009.07. |
| 28. |
笹野 善愛、外園 栄作、大澤 進, 尿中微量蛋白定量の新しい色素法の開発, 生物試料分析科学会 生物試料分析 , 31, 5, 361-368, Vol.31, No5, 361-368, 2008.12. |
| 29. |
Hiroko Ota, Hideto Tamezane, Yoshie Sasano, Eisaku Hokazono, Yuko Yasuda, Shin-ichi Sakasegawa, Shigeyuki Imamura, Tomohiro Tamura, Susumu Osawa, Enzymatic Characterization of an Amine Oxidase from Arthrobacter sp. Used to Measure
Phosphatidylethanolamine., Biosci. Biotecnol. Biochem, 10.1271/bbb.80365, 72, 10, 2732-2738, 72(10), 2732-2738, 2008.10. |
| 30. |
堀田 正敏、杉本 晋哉、外園 栄作、大澤 進, 自己採血による即時血漿分離輸送検査システムの構築 ー 採取量の異なる試料への内部標準による希釈率算定法 ー, 臨床病理, 56, 7, 577-583, Vol.56, No.7, 577-83, 2008.07. |
| 31. |
外園 栄作、中野 倫太、川元 ゆかり、小口 雄二、堀田 多恵子、栢森 裕三、大澤 進, クロロホスフォナゾ−Ⅲを用いた新たな血清カルシウム測定試薬の開発, 生物試料分析科学会 生物試料分析 , 30, 2, 164-171, Vol.30, No2, 164-171, 2007.03. |
| 32. |
横溝佳代 中山亜紀 外園栄作 二ノ宮明子 三宅 瑠璃子 平塚信夫 奥山光彦 加藤祐司 小林静子 伊藤喜久 芝紀代子, 尿路結石患者の結石抽出液と尿のタンパク成分の分析, 日本臨床検査医学会 臨床病理, 53, 12, 1109-1115, 第53巻,第12号,1109-1115, 2005.12. |
| 33. |
三宅瑠璃子, 町井涼子, 二ノ宮明子, 外園栄作, 平塚信夫, 芝紀代子, 伊藤喜久, 山口 聡, 奥山光彦, 加藤祐司, 金子茂男, 尿結石抽出液中の蛋白の同定, 生物物理化学会誌, 48, 21, 48 : 21, 2004., 2004.10. |
| 34. |
外園 栄作、福島 功平、保坂 俊明、芝 紀代子, 全自動電気泳動装置EPA-696を用いた血清蛋白の基礎的検討, 日本臨床自動化学会誌 , 694-696, 2002.Vol 27-5, 2002.03. |
