• 생약학회지
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• 제15권1호
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• pp.39-42
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• 1984

To examine stability and a separate quantitative method of a mixed preparation of lactic acid bacteria, a capsule containing Lactobacillus acidophilus, Lactobacillus bulgaricus and Streptococcus thermophilus was suspended and diluted in sterile water. After the diluted suspension was spread on three media of tryptone glucose extract agar, MRS agar and MRS-sucrose agar, their colonies appeared and were counted. The viable counts exceeded the minimum number of the three bacteria and showed that the mixed preparation was stable at least for 18 months. The results also showed that a separate quantitation of viable cells of the each strain was feasible.

• 한국환경농학회지
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• 제29권3호
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• pp.247-256
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• 2010

A high-performance liquid chromatographic (HPLC) method was developed to determine buprofezin residues in hulled rice and fruits. The buprofezin residue was extracted with acetone and the extract was stepwise purified by liquid-liquid partition and Florisil column chromatography. For rice samples, acetonitrile/n-hexane partition was additionally employed to remove nonpolar lipids. Reversed phase HPLC using an octadecylsilyl column was successfully applied to separate buprofezin from sample co-extractives, as detected by ultraviolet absorption at 250 nm. Recovery experiment at the limit of quantitation validated that the proposed method could evidently determine the buprofezin residue at the level of 0.02 mg/kg. Mean recoveries from hulled rice, apple, pear, and persimmon samples fortified at three tenfold levels were in the range of 80.8~85.2%, 89.1~98.4%, 88.8~95.7% and 90.8~96.2%, respectively. Relative standard deviations of the analytical method were all less than 5%, irrespective of sample types. A selected-ion monitoring LC/mass spectrometry with positive electrospray ionization was also provided to sensitively confirm the suspected residue.

• Archives of Pharmacal Research
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• 제19권6호
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• pp.475-479
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• 1996

Diethylcarbamazine (DEC, 1-diethylcarbamyl-4-methylpiperazine) is an antiparasitic piperazine derivative used in the treatment of lymphatic filariasis caused by Wuchereria bancrofti, Brugia malayi or grugia timori. DEC-N-oxide is a major metabolite in humans and has antifilarial activity. In carrying out pharmacokinetic studies, gas chromatographic analysis of DEC in plasma can be complicated by the presence of the metabolite, since the thermally unstable DEC-N-oxide is converted back to a material which coelutes with DEC under the conditions of the analysis. We now report a method to separate DEC-N-oxide from DEC in plasma using solid phase extraction with subsequent gas chromatographic analysis using a nitrogen specific detector. One-diethylcarbamyl-4-ethylpiperazine (E-DEC) was the internal standard. The standard curve of DEC was linear in the range of 10 to 200 ng/ml as described by Y=0.0350+0.0128X, $R^2=0.999$. The limit of quantitation was 4 ng/mL. Reproducibility at 10, 100 and 200 ng/mL concentration points of the standard curve gave coefficient variations of 6.1%, 7.8% and 1.6%, respectively. The recovery following solid phase extraction was 99.3% for DEC and 94.8% for the internal standard. This sensitive and specific analytical method is suitable for pharmacokinetic studies of DEC.

• 한국환경농학회지
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• 제30권4호
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• pp.432-439
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• 2011

BACKGROUND: An official analytical method was developed to determine etofenprox residues in agricultural commodities using high-performance liquid chromatography (HPLC). METHODS AND RESULTS: The etofenprox residue was extracted with acetone from representative samples of five raw products which comprised rice grain, apple, mandarin, cabbage, and soybean. The extract was then serially purified by liquid-liquid partition and Florisil column chromatography. For rice and soybean samples, acetonitrile/n-hexane partition was additionally coupled to remove nonpolar lipids. Reversed phase HPLC using an octadecylsilyl column was successfully applied to separate etofenprox from co-extractives. Intact etofenprox was sensitively detected by ultraviolet absorption at 225 nm. Recovery experiment at the quantitation limit validated that the proposed method could apparently determine the etofenprox residue at 0.02 mg/kg. Mean recoveries from five crop samples fortified at three levels in triplicate were in the range of 93.6~106.4%. Relative standard deviations of the analytical method were all less than 10%, irrespective of crop types. A selected-ion monitoring LC/mass spectrometry with positive atmospheric-pressure chemical ionization was also provided to confirm the suspected residue. CONCLUSION(s): The proposed method is simple, rapid and sensitive enough to be employed in routine inspection or monitoring of agricultural products for the etofenprox residue.

• Parasites, Hosts and Diseases
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• 제45권4호
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• pp.287-293
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• 2007

The identification and characterization of antigens that elicit human T cell responses is an important step toward understanding of Leishmania major infection and ultimately in the development of a vaccine. Micropreparative SDS-PAGE followed by electro transfer to a PVDF membrane and elution of proteins from the PVDF, was used to separate 2 novel proteins from L. major promastigotes, which can induce antibodies of the IgG2a isotype in mice and also are recognized by antisera of recovered human cutaneous leishmaniasis subjects. Fractionation of the crude extract of L. major revealed that all detectable proteins of interest were present within the soluble Leishmania antigens (SLA). Quantitation of these proteins showed that their expression in promastigotes is relatively very low. Considering the molecular weight, immunoreactivity, chromatographic and electrophoretic behavior in reducing and non-reducing conditions, these proteins are probably 2 isoforms of a single protein. A digest of these proteins was resolved on Tricine-SDS-PAGE and immunoreactive fragments were identified by human sera. Two immunoreactive fragments (36.4 and 34.8 kDa) were only generated by endoproteinase Glu-C treatment. These immunoreactive fragments or their parent molecules may be ideal candidates for incorporation in a cocktail vaccine against cutaneous leishmaniasis.

• 한국축산식품학회지
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• 제41권4호
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• pp.731-738
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• 2021

Herein, a novel analytical method using a high-performance liquid chromatography-fluorescence detector (HPLC/FLD) is developed for rapidly measuring an L-carnitine ester derivative in infant powdered milk. In this study, solid-phase extraction cartridges filled with derivatized methanol and distilled water were used to effectively separate L-carnitine. Protein precipitation pretreatment was carried out to remove the protein and recover the analyte extract with a high recovery (97.16%-106.56%), following which carnitine in the formula was derivatized to its ester form. Precolumn derivation with 1-aminoanthracene (1AA) was carried out in a phosphate buffer using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) as the catalyst. Method validation was performed following the AOAC guidelines. The calibration curves were linear in the L-carnitine concentration range of 0.1-2.5 mg/L. The lower limit of quantitation and limit of detection of L-carnitine were 0.076 and 0.024 mg/L, respectively. The intra- and interday precision and recovery results were within the allowable limits. The results showed that our method helped reduce the sample preparation time. It also afforded higher resolution and better reproducibility than those obtained by traditional methods. Our method is suitable for detecting the quantity of L-carnitine in infant powdered milk containing a large amount of protein or starch.

• 대한화장품학회지
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• 제49권4호
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• pp.291-298
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• 2023

본 연구에서는, 화장품 중 로션, 크림, 클렌저 제형에 적용된 세라마이드엔피(ceramide NP)의 함량을 분석하고자 high-performance liquid chromatography (HPLC)를 이용한 정량 분석 방법을 개발하였다. 분석은 C18 칼럼을 이용하고 이동상은 아세토니트릴과 메탄올을 70 : 30 비율로, 유속은 0.8 mL/min으로 하고 칼럼 온도는 20 ℃로 조건을 설정하였다. 분석 방법은 ICH 가이드라인에 따라 특이성, 직선성, 검출한계(LOD) 및 정량한계(LOQ), 정확성, 정밀성을 분석하여 검증하였다. 분석 방법의 유효성 검증 결과, 검량선의 직선성(R²)은 0.99984로 우수한 직선성을 보였다. 로션, 크림, 클렌저 제형에 대한 정확성은 95.11 ~ 100.48%의 회수율을 통해 확인하였고, 정밀성 분석 결과 상대 표준 편차(RSD)는 0.26% 이하로 나타났다. 검출한계는 0.902 ㎍/mL, 정량한계는 2.733 ㎍/mL로 확인되었다. 이를 통해, 화장품에 적용된 세라마이드엔피의 정량 분석 시, 방해 물질의 영향으로 인해 주피크 분리가 어려운 경우 측정이 가능하게 하여 제품의 품질관리에 도움이 될 것으로 기대된다.

• BMB Reports
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• 제30권6호
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• pp.390-396
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• 1997

A sandwich-type enzyme-linked immunosorbent assay (ELISA) for the quantification of human apolipoprotein A-I (apoA-I) was developed using monoclonal antibodies. For this assay, we used three monoclonal antibodies to trap and detect apo A-I. HDAI16 and HDA15 monoclonal antibodies were used for trapping apoA-I and HDAI8 monoclonal antibody was for detecting apoA-I. These three monoclonal antibodies were produced by immunizing mice with high density lipoprotein (HDL) isolated from human plasma. By immunoblot analysis, these three monoclonal antibodies were specific to apoA-I and showed no cross-reactivities with other plasma proteins. The results of competition assays for epitope cross-reactivity test also verified that these monoclonal antibodies identified separate and distinct epitopes on HDL and apoA-I. Affinity constants of monoclonal antibodies were measured by ELISA. Their association constants ranged from $10^7$ to $10^8$ $M^{-1}$. For this assay, pure apoA-I was isolated by affinity chromatography using monoclonal antibodies. In this sandwich assay, the amount of HRP-labeled HDAI8 bound to apoA-I trapped by HDAI16 and HDAI5 was proportional to apoA-I concentration in the range of 0 to 500ng/ml. ApoA-I concentration in plasma was calculated from the linear regression equation of standard curve. The precision and reliability of the assays are reflected in the low intra-and interassay coefficients of variation that averaged 3.25% and 4.30%, respectively. This assay is sensitive, simple, reproducible, convenient in incubation interval, and does not use radioisotope: thus it can be widely applied in clinical laboratories.

• Journal of Plant Biology
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• 제24권4호
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• pp.217-231
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• 1981

본 연구는 화분단백질을 대상으로 Quercus속 기본종간의 grouping 확인과 그들간의 유사도를 혈청학적 방법으로 결정하고, rocket immunoelectrophoresis의 기술적 가치와 계통분류학적 가치를 논의하였다. 전반적인 정량적, 정성적 data로 보아 Quercus속이 Fagus속과는 멀리 분리되고, Quercus 속내에서는 Cyclobalanopsis 아속이 Lepidobalanus 아속으로부러 뚜렷하게 분리된다. 그러나 Cyclobalanopsis아속이 독립된 속계급까지의 승격은 보증되지 못한다. Lepidobalanus아속에 속하는 종들로 생산된 면역혈청은 같은 아속내의 종들과의 반응에서 강하게 나타났다. 특히 Q. aliena, Q. donarium, Q. serrata는 혈청학적 유사도에 있어서 상호일동으로 나타났다. Q. acutissima는 상기한 종들로부터 약간 멀어진 것 같고, 잡종인 Q. acutissima$\times$variabilis와는 거의 동일성을 나타냈다. 본 계통분류학적 연구에 이용된 rocket immunoelectrophoresis는 새로운 기술로서 그 가치가 증명되었다. 단백질 유사성의 정도를 얻기 위하여 이 기술이 응용되었는데, 모든 rocket height의 총 합계로 결정되었다

• The Korean Journal of Pain
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• 제32권3호
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• pp.168-177
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• 2019

Background: Brennan's rodent paw incision model has been extensively used for understanding mechanisms underlying postoperative pain in humans. However, alterations of physiological parameters like blood pressure and heart rate, or even feeding and drinking patterns after the incision have not been documented as yet. Moreover, though eicosanoids like prostaglandins and leukotrienes contribute to inflammation, tissue levels of these inflammatory mediators have never been studied. This work further investigates the antinociceptive effect of protein C after intra-wound administration. Methods: Separate groups of Sprague-Dawley rats were used for quantitation of cyclooxygenase (COX) activity and leukotriene B4 level by enzyme-linked immunosorbent assay, as well as estimation of cardiovascular parameters and feeding and drinking behavior after paw incision. In the next part, rats were subjected to incision and $10{\mu}g$ of protein C was locally administered by a micropipette. Both evoked and non-evoked pain parameters were then estimated. Results: COX, particularly COX-2 activity and leukotriene B4 levels increased after incision. Hemodynamic parameters were normal. Feeding and drinking were affected on days 1 and 3, and on day 1, respectively. Protein C attenuated non-evoked pain behavior alone up to day 2. Conclusions: Based upon current observations, Brennan's rodent paw incision model appears to exhibit a prolonged period of nociception similar to that after surgery, with minimal interference of physiological parameters. Protein C, which is likely converted to activated protein C in the wound, attenuated the guarding score, which probably represents pain at rest after surgery in humans.