Alleviation of ascorbic acid-induced gastric high acidity by calcium ascorbate in vitro and in vivo

Joon-Kyung Lee,Sang-Hyuk Jung,Sang-Eun Lee,Joo-Hui Han,Eunji Jo,Hyun-Soo Park,Kyung-Sun Heo,Deasun Kim,Jeong-Sook Park,Chang-Seon Myung 대한생리학회-대한약리학회 2018 The Korean Journal of Physiology & Pharmacology Vol.13 No.4

Ascorbic acid is one of the most well-known nutritional supplement and antioxidant found in fruits and vegetables. Calcium ascorbate has been developed to mitigate the gastric irritation caused by the acidity of ascorbic acid. The aim of this study was to compare calcium ascorbate and ascorbic acid, focusing on their antioxidant activity and effects on gastric juice pH, total acid output, and pepsin secretion in an <i>in vivo </i>rat model, as well as pharmacokinetic parameters. Calcium ascorbate and ascorbic acid had similar antioxidant activity. However, the gastric fluid pH was increased by calcium ascorbate, whereas total acid output was increased by ascorbic acid. In the rat pylorus ligation-induced ulcer model, calcium ascorbate increased the gastric fluid pH without changing the total acid output. Administration of calcium ascorbate to rats given a single oral dose of 100 mg/kg as ascorbic acid resulted in higher plasma concentrations than that from ascorbic acid alone. The area under the curve (AUC) values of calcium ascorbate were 1.5-fold higher than those of ascorbic acid, and the C_max value of calcium ascorbate (91.0 ng/ml) was higher than that of ascorbic acid (74.8 ng/ml). However, their T_max values were similar. Thus, although calcium ascorbate showed equivalent antioxidant activity to ascorbic acid, it could attenuate the gastric high acidity caused by ascorbic acid, making it suitable for consideration of use to improve the side effects of ascorbic acid. Furthermore, calcium ascorbate could be an appropriate antioxidant substrate, with increased oral bioavailability, for patients with gastrointestinal disorders.

Alleviation of ascorbic acid-induced gastric high acidity by calcium ascorbate in vitro and in vivo

이준경,정상혁,이상은,한주희,조은지,박현수,허경선,김대선,박정숙,명창선 대한약리학회 2018 The Korean Journal of Physiology & Pharmacology Vol.22 No.1

Ascorbic acid is one of the most well-known nutritional supplement and antioxidant found in fruits and vegetables. Calcium ascorbate has been developed to mitigate the gastric irritation caused by the acidity of ascorbic acid. The aim of this study was to compare calcium ascorbate and ascorbic acid, focusing on their antioxidant activity and effects on gastric juice pH, total acid output, and pepsin secretion in an in vivo rat model, as well as pharmacokinetic parameters. Calcium ascorbate and ascorbic acid had similar antioxidant activity. However, the gastric fluid pH was increased by calcium ascorbate, whereas total acid output was increased by ascorbic acid. In the rat pylorus ligation-induced ulcer model, calcium ascorbate increased the gastric fluid pH without changing the total acid output. Administration of calcium ascorbate to rats given a single oral dose of 100 mg/kg as ascorbic acid resulted in higher plasma concentrations than that from ascorbic acid alone. The area under the curve (AUC) values of calcium ascorbate were 1.5-fold higher than those of ascorbic acid, and the Cmax value of calcium ascorbate (91.0 ng/ml) was higher than that of ascorbic acid (74.8 ng/ml). However, their Tmax values were similar. Thus, although calcium ascorbate showed equivalent antioxidant activity to ascorbic acid, it could attenuate the gastric high acidity caused by ascorbic acid, making it suitable for consideration of use to improve the side effects of ascorbic acid. Furthermore, calcium ascorbate could be an appropriate antioxidant substrate, with increased oral bioavailability, for patients with gastrointestinal disorders.

Cellulomonas sp. AP-7이 생산하는 Ascorbic Acid Phosphorylating Enzyme의 정제 및 특성

이상협,최현일,방원기 한국산업미생물학회 1997 한국미생물·생명공학회지 Vol.25 No.3

Ascorbic acid와 pyrophosphate로부터 ascorbic acid-2-phosphate의 생성을 촉매하는 ascorbic acid phosphorylating enzyme를 DEAE-Sehacel 이온교환크로마토그래피와 Sephacryl S-200 겔여과크로마토그래피를 이용하여 Cellulomonas sp. AP-7의 무세포추출물로부터 32.7-folds로 정제하였다. 정제된 효소의 분자량은 high performance gel chromatography 상에서 96.1 kDa으로 측정되었다. SDS-PAGE 분석에 의해, 본 효소는 24.6 kDa의 동일한 4개의 subunit로 구성되어 있는 것으로 추정하였다. 정제효소의 최적온도와 최적 pH는 각각 40℃와 4.5이었다. Ascorbic acid-2-phosphate 생성반응에 있어서, ascorbic acid와 pyrophosphate에 대한 Km값은 각각 119 mM과 11.9 mM이었다. 본 효소는 5.5'-dithiobis-(2-nitrobenzoic acid)에 의해 활성이 51% 감소하였다. 한편, 본 효소는 phosphatase 활성도 나타내었으며, ascorbic acid-2-phosphate에 대한 Km 값은 7.9 mM이었다. An ascorbic acid phosphorylating enzyme, which catalyzes the formation of ascorbic acid-2-phosphate from ascorbic acid and pyrophosphate, was purified 32.7-folds to homogeneity from a cell-free extract of Cellulomonas sp. Ap-7. The combination of DEAE- Sephacel ion exchange chromatography and Sephacryl S-200 gel filtration was used for their purification. The molecular weight of the native protein was estimated to be 96.1kDa on high performance gel filtration chromatography. The SDS-PAGE analysis indicated that the protein consisted of four identical subunits of 24.6 kDa. The purified enzyme showed the optimal tempeature of 40℃ and optimal pH of 4.5. The Km for ascorbic acid and pyrophosphate were 119 mM and 11.9 mM, respectively. The addition of 5,5'-dithiobis-(2-nitrobenzoic acid) into the reaction mixture resulted in the reduction of the enzyme activity at 51%. The enzyme also had a phosphatase activity at weakly acidic pH and the Km for ascorbic acid-2-phosphate in phosphatase activity was 7.9 mM.

Ascorbic acid insufficiency induces the severe defect on bone formation via the down-regulation of osteocalcin production

Won Kim,Seyeon Bae,Hyemin Kim,Yejin Kim,Jiwon Choi,Sun Young Lim,Hei Jin Lee,Jihyuk Lee,Jiyea Choi,Mirim Jang,Kyoung Eun Lee,Sun G,Chung,Young-il Hwang,Jae Seung Kang,Wang Jae Lee 대한해부학회 2013 Anatomy & Cell Biology Vol.46 No.4

The L-gulono-γ-lactone oxidase gene (Gulo) encodes an essential enzyme in the synthesis of ascorbic acid from glucose. On the basis of previous findings of bone abnormalities in Gulo-/- mice under conditions of ascorbic acid insufficiency, we investigated the effect of ascorbic acid insufficiency on factors related to bone metabolism in Gulo-/- mice. Four groups of mice were raised for 4 weeks under differing conditions of ascorbic acid insufficiency, namely, wild type; ascorbic acid-sufficient Gulo-/- mice, 3-week ascorbic acid-insufficient Gulo-/- mice, and 4-week ascorbic acid-insufficient Gulo-/- mice. Four weeks of ascorbic acid insufficiency resulted in significant weight loss in Gulo-/- mice. Interestingly, average plasma osteocalcin levels were significantly decreased in Gulo-/- mice after 3 weeks of ascorbic acid insufficiency. In addition, the tibia weight in ascorbic acid-sufficient Gulo-/- mice was significantly higher than that in the other three groups. Moreover, significant decreases in trabecular bone volume near to the growth plate, as well as in trabecular bone attachment to the growth plate, were evident in 3- or 4-week ascorbic acid-insufficient Gulo-/-. In summary, ascorbic acid insufficiency in Gulo-/- mice results in severe defects in normal bone formation, which are closely related to a decrease in plasma osteocalcin levels.

Alleviation of ascorbic acid-induced gastric high acidity by calcium ascorbate in vitro and in vivo

Lee, Joon-Kyung,Jung, Sang-Hyuk,Lee, Sang-Eun,Han, Joo-Hui,Jo, Eunji,Park, Hyun-Soo,Heo, Kyung-Sun,Kim, Deasun,Park, Jeong-Sook,Myung, Chang-Seon The Korean Society of Pharmacology 2018 The Korean Journal of Physiology & Pharmacology Vol.22 No.1

Ascorbic acid is one of the most well-known nutritional supplement and antioxidant found in fruits and vegetables. Calcium ascorbate has been developed to mitigate the gastric irritation caused by the acidity of ascorbic acid. The aim of this study was to compare calcium ascorbate and ascorbic acid, focusing on their antioxidant activity and effects on gastric juice pH, total acid output, and pepsin secretion in an in vivo rat model, as well as pharmacokinetic parameters. Calcium ascorbate and ascorbic acid had similar antioxidant activity. However, the gastric fluid pH was increased by calcium ascorbate, whereas total acid output was increased by ascorbic acid. In the rat pylorus ligation-induced ulcer model, calcium ascorbate increased the gastric fluid pH without changing the total acid output. Administration of calcium ascorbate to rats given a single oral dose of 100 mg/kg as ascorbic acid resulted in higher plasma concentrations than that from ascorbic acid alone. The area under the curve (AUC) values of calcium ascorbate were 1.5-fold higher than those of ascorbic acid, and the $C_{max}$ value of calcium ascorbate (91.0 ng/ml) was higher than that of ascorbic acid (74.8 ng/ml). However, their $T_{max}$ values were similar. Thus, although calcium ascorbate showed equivalent antioxidant activity to ascorbic acid, it could attenuate the gastric high acidity caused by ascorbic acid, making it suitable for consideration of use to improve the side effects of ascorbic acid. Furthermore, calcium ascorbate could be an appropriate antioxidant substrate, with increased oral bioavailability, for patients with gastrointestinal disorders.

벼의 ${\alpha}-Glucosidase$에 의한 Ascorbic acid로부터 Ascorbic acid-2-Glucoside의 생산

김성균,황기철,방원기,Kim, Sung-Kyoon,Hwang, Ki-Chul,Bang, Won-Gi 한국응용생명화학회 2000 Applied Biological Chemistry (Appl Biol Chem) Vol.43 No.1

Ascorbic acid로부터 $2-O-{\alpha}-D-glucopyranosyl-L-ascorbic$ acid (AA-2G)를 생산하기 위하여, transglucosylation 활성을 가지는 발아미의 ${\alpha}-glucosidase$를 효소원으로 이용하였다. 시험한 6 품종의 벼중에서 일품벼의 ${\alpha}-glucosidase$ 활성이 125.0 unit/ml로 가장 높았으며, 발아후 3일째에서 최대 비활성 8.52 unit/ml protein을 보였다. 일품벼의 조효소액을 이용한 AA-2G 생산에 있어서 glucose 공여체로는 maltose가 가장 좋았으며, maltose와 ascorbic acid의 최적 농도는 각각 125 mM와 175 mM이었다 ${\alpha}-Glucosidase$ 농도는 100 unit에서 가장 좋았으며, 효과적인 완충용액은 sodium citrate였고, 최적 농도는 100 mM이었다 최적 pH 및 반응은도는 각각 5.0과 $60^{\circ}C$이었다. 상기의 최적 반응조건 하에서 35분 반응후에 ascorbic acid로부터 $108.43\;{\mu}M/unit$의 AA-2G가 생산되었으며, 전환율은 ascorbic acid에 대해 6.2%였다. For the enzymatic production of $2-O-{\alpha}-D-glucopyranosyl-L-ascorbic$ acid (AA-2G) from ascorbic acid, rice seed was used as the source of ${\alpha}-glucosidase$ having transglucosylation activity. Among six rice varieties, cultivated in Korea, ${\alpha}-glucosidase$ activity of Oryza savita L. cv. Ilpumbyeo was the highest with 125.03 unit/ml and it had maximum specific activity with 8.52 unit/mg protein when rice seeds were grown for 3 days after germination. For the production of AA-2G using crude extract of O. savita L. cv. Ilpumbyeo, maltose was most effective glucose donor. The optimum concentration of maltose and ascorbic acid were 125 mM and 175 mM, respectively. The optimum concentration of ${\alpha}-glucosidase$ was 100 unit. The most effective buffer was 100 mM sodium citrate. The optimum pH and temperature were 5.0 and $60^{\circ}C$, respectively. Under the optimum condition, $108.43\;{\mu}M/unit$ of AA-2G was produced from ascorbic acid after 35 minutes of reaction, which corresponds to 6.2% of conversion ratio based on the amount of ascorbic acid used.

Ascorbic acid가 생쥐 복강내 거식세포의 탐식작용과 임파구의 활성에 미치는 영향

이상익,강영준,정가진 大韓免疫學會 1993 大韓免疫學會誌 Vol.15 No.-

Ascorbic acid is the most important water soluble antioxidant as well as a cofactor in mangy hydroxylating reactions. There is a lot of evidence that immune system is sensitive to the level of ascorbic acid, and in our previous report it increased the antibody production of the mouse in vivo In this study we examined the effects of ascorbic acid on the phagocytosis and on the activities of the lymphocytes in mouse system. Murine peritoneal macrophages elicited with caseinate anc treated with ascorbic acid in vivo, showed increased phagocytic activity on the opsonized SRBC bui showed decreased detection of hydrogen peroxide(H202). When treated with ascorbic acid in vitro peritoneal macrophages showed no effect on phagocytosis. To examine the indirect effects of ascorbic acid on phagocytosis via lymphocytes, phorbol myristate acetate(PMA) -treated thymocy tes or T cells were incubated with ascorbic acid and macrophage activating capacity was assessed Culture supernatants of ascorbic acid-treated thymocytes and T cells enhanced the phagocytic activity of macrophages at the concentration of lug/ml. The proliferation of the thymocytes anc the blast formation of B cells were correlated with the level of ascorbic acid. Furthermore, ascorbic acid increased antibody production of splenocytes or B cells in vitro, confirming that it affect; immune system via lymphocytes. With the present results, we can explain in part the broad effect, of ascorbic acid on immune system by its influence on lymphocytes.

Pseudomonas sp.에 의한 Ascorbic acid로부터 Ascorbic acid-2-phosphate의 생산

권기성,이상협,방원기 한국산업미생물학회 2000 한국미생물·생명공학회지 Vol.28 No.1

Ascorbic acid와 인산기 공여체로부터 ascorbic acid-2-phosphate를 생산하기 위하여 ascorbic acid를 ascorbic acid-2-phosphate로 전환할 수 있는 세균을 토양과 본 실험실 보관 균주로부터 분리하였다. 이들 중 ascorbic acid-2-phosphate 생산능이 가장 좋은 LSH-3를 선별하였으며 Pseudomonas sp.로 부분 동정하였다. Pseudomonas sp. LSH-3의 휴지세포를 효소원으로 사용하여 ascorbic acid와 인산기 공여체로부터 ascorbic acid-2-phosphate를 생산하는 최적 반응조건을 검토하였다. 상기의 결과들을 요약하면 다음과 같다. Ascorbic acid-2-phosphate를 생산하기위한 최적 배양시간과 균체농도는 14시간, 100g/l(wet weight)이었으며 0.1%(v/v) Triton X-100이 가장 효과적인 유화제이었다. Ascorbic acid와 pyrophosphate의 최적 농도는 각각 400 mM과 500 mM이었으며 이때 14.54g/l의 ascorbic acid-2-phosphate가 생산되었다. 가장 효과적인 완충용액은 50 mM sodium acetate이었다. 최적 pH 및 온도는 각각 4.5와 40℃이었으며 위의 조건하에서 32시간 반응 후에 ascorbic acid로부터 17.71g/l의 ascorbic acid-2-phosphate가 생산되었으며 전환율은 ascorbic acid에 대해 17.5%이었다. In order to produce ascorbic acid-2-phosphate from ascorbic acid, bacteria capable of transforming ascorbic acid to ascorbic acid-2-phosphate were isolated from soils and the stock cultures in our laboratory. Among them, a newly isolated bacterium LSH-3 having the best ability of producing ascorbic acid-2-phosphate was selected and partially identified as Pseudomonas sp. The optimum conditions for the production of ascorbic acid-2-phosphate from ascorbic acid and using its resting cells as the source of enzyme were investigated. the results were summarized as follows: The optimum cultivation time and the cell weight for the production of ascorbic acid-2-phosphate was 14 hours and 100 g/l(wet weight), respectively. And 0.1%(v/v) Trition X-100 was the most effective surfactant. The optimum concentrations of ascorbic acid and pyrophosphate were 400 mM and 500 mM, respectively, which led to produce 14.54 g/l of ascorbic acid-2-phosphate. The most effective buffer was 50 mM sodium acetate. The optimum pH and temperature were 4.5 and 40℃, respectively. Under the above conditions, 17.71 g/l of ascorbic acid-2-phosphate was produced from ascorbic acid after 32 hour-incubation, which corresponded to 17.5% of conversion rate based on ascorbic acid.

The Microencapsulated Ascorbic Acid Release in vitro and Its Effect on Iron Bioavailability

Lee, Jun-Beum,Ahn, Joung-Jwa,Lee, Jong-Hwi,Kwak, Hae-Soo The Pharmaceutical Society of Korea 2003 Archives of Pharmacal Research Vol.26 No.10

The present study was carried out to examine the stability of microencapsulated ascorbic acid in simulated-gastric and intestinal situation in vitro and the effect of microencapsulated ascorbic acid on iron bioavailability. Coating materials used were polyglycerol monostearate (PGMS) and medium-chain triacylglycerol (MCT), and core materials were L-ascorbic acid and ferric ammonium sulfate. When ascorbic acid was microencapsulated by MCT, the release of ascorbic acid was 6.3% at pH 5 and 1.32% at pH 2 in simulated-gastric fluids during 60 min. When ascorbic acid was microencapsulated by PGMS, the more ascorbic acid was released in the range of 9.5 to 16.0%. Comparatively, ascorbic acid release increased significantly as 94.7% and 83.8% coated by MCT and PGMS, respectively, for 60 min incubation in simulated-intestinal fluid. In the subsequent study, we tested whether ascorbic acid enhanced the iron bioavailability or not. In results, serum iron content and transferring saturation increased dramatically when subjects consumed milks containing both encapsulated iron and encapsulated ascorbic acid, compared with those when consumed uncapsulated iron or encapsulated iron without ascorbic acid. Therefore, the present data indicated that microencapsulated ascorbic acid with both PGMS and MCT were effective means for fortifying ascorbic acid into milk and for enhancing the iron bioavailability.

The Role of Ascorbic Acid on the Redox Status and the Concentration of Malondialdehyde in Streptozotocin-Induced Diabetic Rats

Choi, Hee-Jung,Je, Hyun-Dong,Jeong, Ji-Hoon,Min, Young-Sil,Choi, Tae-Sik,Park, Joon-Hong,Shin, Chang-Yell,Sohn, Uy-Dong The Pharmaceutical Society of Korea 2003 Archives of Pharmacal Research Vol.26 No.3

We investigated the role of ascorbic acid on the redox status in streptozotocin-induced diabetic rats. In the plasma of diabetic rats, the ratio of reduced/total ascorbic acid was significantly decreased as compared with normal control. Ascorbic acid supplementation increased the reduced and total ascorbic acid contents as compared with diabetic control. In the rutintreatment group, reduced and total contents of ascorbic acid were significantly decreased, however, the ratio of reduced/total contents of ascorbic acid had no difference as compared with diabetic rats. In the insulin-treatment group, this ratio is not significantly different as compared with diabetic control. However, in the insulin plus ascorbic acid treatment group, reduced form and the ratio of reduced/total ascorbic acid were significantly increased as compared with diabetic control. In addition, we measured the contents of malondialdehyde (MDA) in the plasma of diabetic rats. The contents of MDA was increased as compared with normal control, however, in insulin-treatment group, the contents of MDA was decreased as compared with diabetic rats. Ascorbic acid had no effects on the increases of MDA in diabetic rats. In conclusion, plasma ascorbic acid level and its reduced/total ratio reflects the status of the oxidative stress in the diabetic rats. Supplement of ascorbic acid did not correct the ratio of the reduced/total ascorbic acid. However, supplement of insulin and ascorbic acid corrected the ratio of reduced/total ascorbic acid.