High-performance liquid chromatographic assay of human lymphocyte kynureninase activity levels
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ObjectiveComparison the two high performance liquid chromatography(HPLC) for serum phenylalanine(phe). MethodsHPLC-ultraviolet absorbing detection(HPLC-UV) and HPLC-fluorescence detection(HPLC-FLD) were compared in linear range, recovery rate detection limit and precision. Identical samples were detected with two HPLC methods, The results were analyzed. ResultsThe linear range, mean recovery rate, in-run CV and between-run CV were 61512 μmol/L, 0.75 μmol/L, 3.68% and 4.01% for HPLC-UV, respectively, and 121220 μmol/L, 3 μmol/L, 102.5%, 4.07% and 7.17% for HPLC-FLD, respectively. ConclusionBoth HPLC are sensitive, accurate, simple, rapid and adapted for clinical use. The HPLC-UV is more presision and rapid than the HPLC-FLD.
Linear range
Recovery rate
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Monolithic HPLC column
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본 연구에서는 UPLC를 이용한 밀크씨슬추출물의 지표 물질인 실리마린의 신규 분석법을 확립하였으며, 건강기능식품공전의 HPLC 분석법과 비교하여 UPLC 분석법을 검증하였다. UPLC를 이용한 신규 분석법을 검증은 특이성, 직선성, 정확성, 정밀성, 실험실 내 정밀성, 완건성 등 6개의 검증 항목을 선정하여 실시하였다. HPLC 및 UPLC 분석법 모두에서 6가지 실리마린 표준물질이 선택적이며 특이적으로 정확하게 검출됨을 확인하였으며, 실리마린 25-400 μg/mL의 농도에서 결정계수 값이 0.9999이상으로 높은 직선성을 갖음을 확인하였다. 정확성에서 UPLC를 이용한 신규 분석법의 회수율은 평균 99%이상으로 HPLC분석법보다 높은 회수율을 보였으며, UPLC 분석법의 회수율편차가 HPLC 분석의 회수율편차의 절반 이하임을 확인하였다. UPLC 분석법에서 얻어진 피크 면적의 상대표준편차가 HPLC 분석법의 그것과 비교하여 절반 이하였으며, UPLC 분석법의 정밀성 및 실험실 내 정밀성이 HPLC 분석법의 정밀성 및 실험실 내 정밀성보다 높음을 확인하였다. 분석법 검증 결과, 모든 검증 항목에서 HPLC와 UPLC 분석법 모두 적합하게 검증되었으며, 검증 항목 중 정확성, 정밀성, 실험실 내 정밀성 및 완건성 항목에서 UPLC 분석법의 검증값이 HPLC 분석법보다 뛰어났다. HPLC 분석법으로 검체 100개를 분석할 경우, 총 분석시간은 95시간 30분 (기기 안정화 시간 포함한 분석시간은 약 96 시간), 총 이동상 소모량은 6,215 mL이고 이동상 소모량 중 methanol 함량은 2,450 mL이었으나, UPLC 분석법의 총 분석시간은 약 13시간 (기기 안정화 시간 포함한 분석시간은 약 14시간), 총 이동상 소모량은 168 mL이며 이동상 소모량 중 methanol 함량은 74 mL에 불과하였다. 이상의 결과를 통해 UPLC를 이용한 실리마린 신규 분석법은 건강기능식품의 실리마린을 분석하는데 적합하며, 건강기능식품공전의 HPLC 분석법과 비교하여 효율적인 분석법이라고 판단할 수 있다.
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A simple, rapid method for the determination of the benzodiazepines, diazepam and chlordiazepoxide, and their metabolites by high performance liquid chromatography (HPLC) has been developed. The procedure is applicable to the assay of other similar drugs in biological fluids. The method utilizes BondElut™ extraction columns to facilitate the extraction. BondElut columns selectively adsorb the benzodiazepines and metabolites from serum at a pH of 9.0. The compounds are eluted with 300μl of methanol which makes sample concentration rapid, if even necessary. Analysis is performed using isocratic reversed-phase chromatography, and quantitation is carried out by ultraviolet (UV) detection. Using this procedure, it is possible to determine drug and metabolite levels to as low as 25 ng/ml in 0.5 ml of serum.
Sample Preparation
Chlordiazepoxide
Solid phase extraction
Oxazepam
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چکیده: زمینه و هدف: گرفتگی عضله انقباض دردناک غیر ارادی و موضعی یک عضله یا قسمتی از یک عضله است. بین 5 تا بیش از 30 زنان از گرفتگی عضلات پا حین حاملگی شکایت دارند. منیزیم رل مهمی در متابولیسم و فعالیت عضلانی بر عهده دارد، بدلیل نبودن ترکیب مناسبی از منیزیم در ایران و ارتباط و تداخل عملکرد منیزیم و ویتامین های B1 و B6 و همینطور بی خطر بودن مصرف ویتامین ها در حین حاملگی، این مطالعه با هدف بررسی اثر درمانی ترکیب ویتامین B1 و B6 روی گرفتگی عضلات پا حین حاملگی انجام گرفت. روش بررسی: در این کار آزمایی بالینی آینده نگر دو سوکور 63 زن مبتلا به گرفتگی عضلات پا از نظر تعداد، شدت درد و زمان وقوع گرفتگی مورد بررسی قرار گرفتند. سپس به آنها به صورت تصادفی یک قوطی محتوی 56 کپسول حاوی 250 میلی گرم ویتامینB1 و 240 میلی گرم ویتامینB6 و یا 56 کپسول مشابه محتوی نشاسته داده شد و به ایشان گفته شد برای چهار هفته روزانه دو کپسول میل کنند. سپس بیماران مجددا از نظر تعداد، شدت درد و زمان وقوع گرفتگی و ارزیابی آنها از اثر درمان بررسی شدند. یافته ها: پس از اتمام درمان تعداد و شدت درد گرفتگی عضلات پا در گروه دارو به طور معنی داری از گروه دارو نما کمتر بود (05/0>p). ولی تفاوت معنی دار بین دو گروه در زمان وقوع گرفتگی ها دیده نشد. در این مطالعه هیچ عارضه جانبی با مصرف دارو یا دارو نما دیده نشد. نتیجه گیری: به نظر می رسد که تجویز ترکیب ویتامین های B1 و B6 در کاهش تعداد و شدت درد گرفتگی عضلات پا حین حاملگی مؤثر باشد.
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We selected 3 kinds of water soluble vitamins such as vitamin B₄, vitamin B₆, and vitamin C using Sprague Dawley(SD) rat model. The concentration of vitamin B₄and vitamin B₆ for sucrose loading test was 50 mg/kg, vitamin C was 0.1 and 0.5 g/kg. The effect of as vitamin B₄, vitamin B₆, and vitamin C on the postprandial blood glucose increase after meal was investigated in SD rats fed on sucrose. Three vitamin samples significantly reduced the postprandial blood glucose levels after sucrose loading. These results indicate that vitamin B₄, vitamin B₆, and vitamin C may have anti‐hyperglycemic effect by suppressing carbohydrate absorption in the gastrointestinal level, and thereby reducing the postprandial increase of blood glucose. These results indicate that vitamin B₄, vitamin B₆, and vitamin C may improve postprandial in blood glucose and glucose homeostasis since it inhibits intestinal sucrase and thus delays carbohydrate absorption, although clinical trials are needed.
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Objective To compare the results of HbAlc in blood samples with various configuration of hemoglobin(Hb) by affinity chromatogragh-high performance liquid chromatography(AC-HPLC)system and ion exchange-high performance liquid chromatography(IE-HPLC)system,and evaluate the effect of glycohemoglobin variants on AC-HPLC and IE-HPLC.MethodsThe levels of HbA1c in 30 blood samples with the normal Hb configuration and the HbA1c levels were 5%-11%,the blood samples with different HbF levels and the blood samples of patients with α-Mediterranean anemia(α-MA)and β-Mediterranean anemia(β-MA) were determined by AC-HPLC and IE-HPLC,respectively.ResultsThere was not difference between the AC-HPLC and IE-HPLC for the blood sanples with normal Hb configuration and the HbA1c levels were 5%-11%(P0.05).There was not interference with HbF for HbA1c determation by AC-HPLC.However,the bias of HbA1c determination by IE-HPLC was occurred when HbF level 8.8%,and the HbA1c was undetectable when HbF level was 70%.For blood samples with patients with α-MA and mitis β-MA,the results of HbA1c deteremind by IE-HPLC were significantly higher than AC-HPLC(P0.01 and P0.05,respectively).The level of HbA1c in patient with gravis type β-MA was undetected by IE-HPLC.ConclusionsThe results of HbA1c in blood samples containing Hb variants or HbF may be interfered when analyzed by IE-HPL.However,there was not interferences with AC-HPLC.
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Bacitracin
Chromatographic Separation
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雄ラットを高たん白ビタミンB6補足自由摂食群と絶食群, B6欠乏自由摂食群と絶食群の4群に分け, B6欠乏の影響, および摂食条件のちがいが, 肝脂質および肝脂肪酸組成にどのような影響を与えるかを検討した結果, つぎのような成績を得た。1) B6欠乏ラットは, B6補足ラットにくらべて, かなり成長が阻害されており, 体重あたりの肝重量が大きく, また, 肝脂質含量もB6欠乏ラットにおいて, B6補足ラットのほぼ2倍に上昇し, グリセライドとコレステロールの増量を伴う脂肪肝を呈した。 B6補足ラットの場合, 19時間絶食にすると肝重量が有意に減少し, また, 肝総脂質量が1/2に減少したが, 肝脂質含量 (%) には変化がみられなかった。 これに対して, B6欠乏ラットは, 19時間絶食にすると肝重量, 肝総脂質量, 肝脂質含量 (%) がやや増加傾向を示したが, 有意差はみられなかった。2) 肝脂肪酸組成は, B6補足ラットの自由摂食群と絶食群を比較すると, ほとんどの脂肪酸において, 有意な変化を示した。 すなわち, C16: 0, C16: 1, C18: 1は有意に減少し, C18: 0, C18: 2, C20: 4は有意な増加を示した。 B6欠乏ラットの場合, いずれの脂肪酸においても, 絶食による有意な変化はみられなかった。 このことは, B6欠乏時には, 脂肪酸の酸化が障害されているのではないかということを示唆している。 なお, アラキドン酸は, B6補足ラットの絶食時にのみ有意に高い値を示している。 paired feedingの場合, B6補足ラットは常に空腹状態に陥っているために, 正常状態に比してアラキドン酸が増加し, その結果, B6欠乏ラットのアラキドン酸含量との間に差が生じる結果を招いている可能性がある。3) 肝血液を除く全体脂肪の脂肪酸組成については, B6欠乏ラットのアラキドン酸含量が著明に高値を示した。
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