JPS6350665B2 - - Google Patents
Info
- Publication number
- JPS6350665B2 JPS6350665B2 JP13847681A JP13847681A JPS6350665B2 JP S6350665 B2 JPS6350665 B2 JP S6350665B2 JP 13847681 A JP13847681 A JP 13847681A JP 13847681 A JP13847681 A JP 13847681A JP S6350665 B2 JPS6350665 B2 JP S6350665B2
- Authority
- JP
- Japan
- Prior art keywords
- reaction
- free cholesterol
- analysis
- sodium
- container
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims description 68
- 238000006243 chemical reaction Methods 0.000 claims description 50
- 235000012000 cholesterol Nutrition 0.000 claims description 34
- 238000004458 analytical method Methods 0.000 claims description 25
- 239000003153 chemical reaction reagent Substances 0.000 claims description 19
- 229920003002 synthetic resin Polymers 0.000 claims description 14
- 239000000057 synthetic resin Substances 0.000 claims description 14
- 108010013563 Lipoprotein Lipase Proteins 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 7
- BCKXLBQYZLBQEK-KVVVOXFISA-M Sodium oleate Chemical compound [Na+].CCCCCCCC\C=C/CCCCCCCC([O-])=O BCKXLBQYZLBQEK-KVVVOXFISA-M 0.000 claims description 5
- RYYKJJJTJZKILX-UHFFFAOYSA-M sodium octadecanoate Chemical compound [Na+].CCCCCCCCCCCCCCCCCC([O-])=O RYYKJJJTJZKILX-UHFFFAOYSA-M 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- BTURAGWYSMTVOW-UHFFFAOYSA-M sodium dodecanoate Chemical compound [Na+].CCCCCCCCCCCC([O-])=O BTURAGWYSMTVOW-UHFFFAOYSA-M 0.000 claims description 3
- 229940082004 sodium laurate Drugs 0.000 claims description 3
- 229940045870 sodium palmitate Drugs 0.000 claims description 3
- 229940080350 sodium stearate Drugs 0.000 claims description 3
- GGXKEBACDBNFAF-UHFFFAOYSA-M sodium;hexadecanoate Chemical compound [Na+].CCCCCCCCCCCCCCCC([O-])=O GGXKEBACDBNFAF-UHFFFAOYSA-M 0.000 claims description 3
- 238000012360 testing method Methods 0.000 claims description 3
- 102100022119 Lipoprotein lipase Human genes 0.000 claims 1
- RLFWWDJHLFCNIJ-UHFFFAOYSA-N 4-aminoantipyrine Chemical compound CN1C(C)=C(N)C(=O)N1C1=CC=CC=C1 RLFWWDJHLFCNIJ-UHFFFAOYSA-N 0.000 description 10
- 239000000523 sample Substances 0.000 description 10
- 102000043296 Lipoprotein lipases Human genes 0.000 description 9
- AZQWKYJCGOJGHM-UHFFFAOYSA-N 1,4-benzoquinone Chemical compound O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 8
- 238000005259 measurement Methods 0.000 description 7
- 210000002966 serum Anatomy 0.000 description 7
- 238000002835 absorbance Methods 0.000 description 6
- -1 alkali metal salts Chemical class 0.000 description 6
- 235000014113 dietary fatty acids Nutrition 0.000 description 6
- 229930195729 fatty acid Natural products 0.000 description 6
- 239000000194 fatty acid Substances 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 5
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 229910052783 alkali metal Inorganic materials 0.000 description 4
- 108010089254 Cholesterol oxidase Proteins 0.000 description 3
- 102000003992 Peroxidases Human genes 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 108040007629 peroxidase activity proteins Proteins 0.000 description 3
- 102000004895 Lipoproteins Human genes 0.000 description 2
- 108090001030 Lipoproteins Proteins 0.000 description 2
- 239000012295 chemical reaction liquid Substances 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 239000000113 methacrylic resin Substances 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 229920005990 polystyrene resin Polymers 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- NYOXRYYXRWJDKP-GYKMGIIDSA-N cholest-4-en-3-one Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 NYOXRYYXRWJDKP-GYKMGIIDSA-N 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 108010090622 glycerol oxidase Proteins 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000005375 photometry Methods 0.000 description 1
- 229920003217 poly(methylsilsesquioxane) Polymers 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Urology & Nephrology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Endocrinology (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Description
本発明は遊離コレステロールを分析する方法に
係り、特に同じ反応容器を洗浄再生して複数種の
被検項目を分析する場合に適用するに好適な遊離
コレステロール分析法に関する。
臨床用生化学分析装置のうち、多種項目を分析
する装置の大部分は、反応ライン系に分析すべき
項目数と同数の反応容器列を備えたものであり、
各反応容器列が各々の分析項目に対応している。
だからこの場合、同じ反応容器に着目すれば、試
料と試薬との反応液を測定に供したあとの反応容
器が洗浄されて再び別の試料の反応のために使わ
れるとしても、同一種の分析項目に関してだけで
ある。
ところが、このような方式の分析装置は、分析
項目の数を増大するにつれて大形化せざるを得な
い。この点を改善したのが、１つの反応容器列上
で複数種の項目を反応させる方式の分析装置であ
る。この種装置では、反応に用いた容器を洗浄し
て再生使用するときに、前の分析項目と後の分析
項目の種類が異なる。
一方、反応容器の材質としてよく用いられるの
は、ガラスおよび合成樹脂である。この内ガラス
製容器は耐熱性があるので洗浄、乾燥して再生使
用する場合に用いられ、合成樹脂製容器は一度の
反応に使用しただけで使い捨てる場合に用いられ
ていた。
最近の技術の進歩により、合成樹脂製容器を洗
浄して再生使用できる可能性が出てきた。それ故
発明者らは同じ反応容器列上で複数種の分析項目
を反応させる分析装置に合成樹脂製容器を採用す
ることを試みた。合成樹脂製容器はガラス製のも
のに比べて軽量であるという利点を有する。
ところが、合成樹脂製容器を洗浄再生使用して
血清試料中の遊離コレステロールを反応させ、吸
光光度法で測定すると、著しい誤差を生ずること
のあることが見い出された。
本発明の目的は、合成樹脂製容器を用いて他の
分析項目用試料を処理したあとに、遊離コレステ
ロール分析反応を生ぜしめても、測定誤差が生じ
ない遊離コレステロール分析法を提供することに
ある。
本発明は、上述の目的を達成するために次の方
法が実行される。すなわち、本発明は、遊離コレ
ステロール以外の被検項目の分析に使用した合成
樹脂製反応容器を洗浄して使用すること、前記洗
浄によつてもリポプロテインリパーゼが除去され
ずに該反応容器内に残存している場合、オレイン
酸ナトリウム、ステアリン酸ナトリウム、ラウリ
ル酸ナトリウム、およびパルミチン酸ナトリウム
からなる群から選ばれた化合物を含む遊離コレス
テロール用分析試薬を試料と混合し、該試薬と該
試料とからなる反応液内での前記化合物の濃度を
２〜200μMとなすこと、および前記反応液を光
学的に測定して遊離コレステロール濃度を求める
ことを含む方法である。
本発明は、合成樹脂製容器を用いて遊離コレス
テロール分析反応を生ぜしめたときの測定誤差の
原因が、合成樹脂製容器、例えばメタクリル樹脂
製容器やポリスチレン樹脂製容器内に残存されて
いるリポプロテインリパーゼの作用によるもので
あることを発見したことに基づいてなされた。リ
ポプロテインリバーゼは、総コレステロール分析
用試薬や中性脂肪分析用試薬に含まれており、生
体試料例えば血清試料中のエステル形コレステロ
ールを遊離コレステロールに転換する働きを有す
る。だから、リポプロテインリパーゼが容器内に
残存すれば、試料の遊離コレステロール濃度が真
値より高濃度となり、測定値に正誤差をもたらす
ものと推定される。このリポプロテインリパーゼ
は容器が合成樹脂製である場合にその容器に物理
的に吸着し、一旦吸着したものは水洗によつて容
易に除去できない。
発明者らは実験を重ねた結果、脂肪酸のアルカ
リ金属塩がリポプロテインリパーゼの活性を妨げ
ることを見い出した。だから、遊離コレステロー
ル分析反応時に脂肪酸アルカリ金属塩を共存させ
れば、反応容器にリポプロテインリパーゼが吸着
されていてもエステル形コレステロールが遊離コ
レステロールに変換されない。
遊離コレステロール分析用試薬に他の物質を共
存させる場合は、測定反応を妨害するものであつ
てはならない。測定反応を妨害せずにリポプロテ
インリパーゼの活性を妨げるために有効な脂肪酸
アルカリ金属塩は、オレイン酸ナトリウム、ステ
アリン酸ナトリウム、ラウリル酸ナトリウム、パ
ルミチン酸ナトリウム等の高級脂肪酸ナトリウム
塩である。反応容器内における脂肪酸アルカリ金
属塩の共存適正量は２〜200μMである。
第１表に本発明を実行するために用いた遊離コ
レステロール分析用試薬液の組成例を示す。
The present invention relates to a method for analyzing free cholesterol, and particularly to a method for analyzing free cholesterol that is suitable for use when a plurality of test items are analyzed by washing and regenerating the same reaction vessel. Among clinical biochemistry analyzers, most of the devices that analyze a wide variety of items are equipped with a reaction line system containing the same number of reaction vessels as the number of items to be analyzed.
Each reaction container row corresponds to each analysis item.
Therefore, in this case, if we focus on the same reaction vessel, even if the reaction vessel is cleaned and used again for the reaction of another sample after the reaction liquid of the sample and reagent is subjected to measurement, it is possible to analyze the same type of reaction vessel. It's only about the item. However, the analyzer of this type has to become larger as the number of analysis items increases. An improvement on this point is an analyzer that allows multiple types of items to react on one row of reaction vessels. In this type of apparatus, when a container used in a reaction is washed and reused, the types of previous analysis items and subsequent analysis items are different. On the other hand, glass and synthetic resin are often used as materials for reaction vessels. Among these, glass containers are heat resistant and are used when they can be recycled after being washed and dried, while synthetic resin containers are used when they are used for a single reaction and then discarded. Recent advances in technology have created the possibility of cleaning and recycling synthetic resin containers. Therefore, the inventors attempted to employ synthetic resin containers in an analyzer that reacts multiple types of analysis items on the same reaction container row. Containers made of synthetic resin have the advantage of being lighter than containers made of glass. However, it has been found that when a synthetic resin container is washed and reused to react with free cholesterol in a serum sample and measured by spectrophotometry, a significant error may occur. An object of the present invention is to provide a free cholesterol analysis method that does not cause measurement errors even if a free cholesterol analysis reaction occurs after processing samples for other analysis items using a synthetic resin container. In order to achieve the above object, the present invention implements the following method. In other words, the present invention provides a method for cleaning a synthetic resin reaction vessel used for analysis of test items other than free cholesterol, and for lipoprotein lipase to remain in the reaction vessel without being removed even after the washing. If any remains, an analytical reagent for free cholesterol containing a compound selected from the group consisting of sodium oleate, sodium stearate, sodium laurate, and sodium palmitate is mixed with the sample and the reagent and the sample are separated from each other. This method includes adjusting the concentration of the compound in the reaction solution to 2 to 200 μM, and optically measuring the reaction solution to determine the free cholesterol concentration. The present invention discloses that the cause of measurement error when performing a free cholesterol analysis reaction using a synthetic resin container is lipoproteins remaining in the synthetic resin container, such as a methacrylic resin container or a polystyrene resin container. This was based on the discovery that this is due to the action of lipase. Lipoprotein revertase is included in total cholesterol analysis reagents and neutral fat analysis reagents, and has the function of converting ester cholesterol in biological samples such as serum samples to free cholesterol. Therefore, if lipoprotein lipase remains in the container, it is estimated that the free cholesterol concentration of the sample will be higher than the true value, causing a positive error in the measured value. This lipoprotein lipase physically adsorbs to the container when the container is made of synthetic resin, and once adsorbed, it cannot be easily removed by washing with water. As a result of repeated experiments, the inventors discovered that alkali metal salts of fatty acids inhibit the activity of lipoprotein lipase. Therefore, if a fatty acid alkali metal salt is present during the free cholesterol analysis reaction, ester cholesterol will not be converted to free cholesterol even if lipoprotein lipase is adsorbed in the reaction vessel. If other substances are present in the reagent for free cholesterol analysis, they must not interfere with the measurement reaction. Fatty acid alkali metal salts effective for inhibiting the activity of lipoprotein lipase without interfering with the measurement reaction are higher fatty acid sodium salts such as sodium oleate, sodium stearate, sodium laurate, and sodium palmitate. The appropriate coexistence amount of fatty acid alkali metal salt in the reaction vessel is 2 to 200 μM. Table 1 shows an example of the composition of the reagent solution for free cholesterol analysis used to carry out the present invention.
【表】
分析用試薬は、コレステロールオキシダーゼと
キノン系発色剤（ここではフエノールおよび４―
アミノアンチピリン）と緩衝剤を含む従来から用
いられていたものに、適量の高級脂肪酸塩が混合
されている。
実施例 １
透明なメタクリル樹脂からなる一列の反応容器
列を間欠移送する間に、第１の場所でサンプラー
からの血清試料をピペツタによつて反応容器内へ
所定量分配し、第２の場所で試薬デイスペンサに
よつて所定量の試薬液を添加し、第３の場所で透
明な反応容器に光を照射し容器内の反応液の吸光
度を測定する臨床用生化学分析装置を用いる。こ
の装置の反応ライン上では、１つの検体に関連す
る複数の分析項目用試料が連続的に配列されたあ
と、他の検体に関連する複数の分析項目用の一連
の試料が配列される。各分析項目の反応液を測定
したあと、反応容器は水洗され、上述の第１の場
所まで移送されその後の試料の反応に供される。
試料を分取した特定の反応容器に、リポプロテ
インリパーゼ、リン酸緩衝液、フエノール、４―
アミノアンチピリン、グリセロールオキシダーゼ
およびパーオキシダーゼを含む中性脂肪分析用試
薬を加えて試料と混合し、所定時間後に反応液の
吸光度測定がなされる。その後この反応容器を水
洗する。水洗された特定の反応容器に、コントロ
ール血清（モニトロール）20μを採取し、
その後第１表の遊離コレステロール分析用試薬液
を200μ添加し、最終液量を１mlにする。
PH7.8の緩衝液中で試料の遊離コレステロール
がコレステロールオキシダーゼにより酸化され、
過酸化水素とΔ4―コレステノンを生ずる。生成
された過酸化水素にパーオキシダーゼの存在下で
フエノールおよび４―アミノアンチピリンを縮合
させて赤色キノンを生成する。光度計によつて反
応液中の赤色キノンに基づく吸光度を測定して遊
離コレステロール濃度を求める。コントロール血
清の遊離コレステロールの標準濃度は45mg／ｄ
であるが、実測値は43〜46mg／ｄであつた。
第１表における試薬液の成分のうち、オレイン
酸ナトリウムの濃度を変えたときのコントロール
血清（モニトロール）中の遊離コレステロー
ルの測定誤差（正誤差）を第２表に示す。[Table] The analytical reagents are cholesterol oxidase and a quinone coloring agent (here, phenol and 4-
An appropriate amount of a higher fatty acid salt is mixed with the conventionally used ingredients, including aminoantipyrine (aminoantipyrine) and a buffering agent. Example 1 During intermittent transfer of a row of reaction vessels made of transparent methacrylic resin, a predetermined amount of serum sample from a sampler is dispensed into the reaction vessels at a first location by a pipette, and at a second location. A clinical biochemical analyzer is used that adds a predetermined amount of a reagent solution using a reagent dispenser, irradiates a transparent reaction container with light at a third location, and measures the absorbance of the reaction solution in the container. On the reaction line of this device, samples for a plurality of analysis items related to one specimen are sequentially arranged, and then a series of samples for a plurality of analysis items related to other specimens are arranged. After measuring the reaction solution for each analysis item, the reaction container is washed with water and transported to the above-mentioned first location, where it is subjected to subsequent sample reactions. Lipoprotein lipase, phosphate buffer, phenol, 4-
A neutral fat analysis reagent containing aminoantipyrine, glycerol oxidase, and peroxidase is added and mixed with the sample, and after a predetermined period of time, the absorbance of the reaction solution is measured. The reaction vessel is then washed with water. Collect 20μ of control serum (Monitrol) into a specific reaction container that has been washed with water.
Then, add 200μ of the reagent solution for free cholesterol analysis shown in Table 1 to bring the final volume to 1ml. Free cholesterol in the sample is oxidized by cholesterol oxidase in a pH 7.8 buffer,
Generates hydrogen peroxide and Δ 4 -cholestenone. The produced hydrogen peroxide is condensed with phenol and 4-aminoantipyrine in the presence of peroxidase to produce a red quinone. The free cholesterol concentration is determined by measuring the absorbance based on red quinone in the reaction solution using a photometer. The standard concentration of free cholesterol in control serum is 45 mg/d.
However, the actual value was 43 to 46 mg/d. Among the components of the reagent solution in Table 1, Table 2 shows the measurement errors (correct errors) of free cholesterol in the control serum (Monitrol) when the concentration of sodium oleate was changed.
【表】
第２表から理されるように、オレイン酸ナトリ
ウムを含まない従来の試薬を用いた場合、中性脂
肪反応後の反応容器を洗浄して遊離コレステロー
ルを反応させて測定すると、標準濃度が45mg／ｄ
であるのに対して140mg／ｄ以上もの測定値
となる。
実施例 ２
ポリスチレン樹脂製の透明な反応容器の列を有
する分析装置を用いた。１つの反応容器列上で複
数種の分析項目を反応させ、直接測光法によつて
反応液の吸光度を測定するのは実施例１の分析装
置と同様である。
分析装置の特定の容器に、総コレステロール用
試料液をサンプラーから採取し、リポプロテイン
リパーゼ、リン酸緩衝液、フエノール、４―アミ
ノアンチピリン、コレステロールオキシダーゼお
よびパーオキシダーゼを含む総コレステロール試
薬液を加えて、反応せしめ、所定時間後に反応液
の吸光度を測定する。その後この反応容器を水洗
し、遊離コレステロールの分析反応に供する。
水洗された特定の反応容器にコントロール血清
を20μ採取し、その後第１表のオレフイン酸ナ
トリウムの代わりに20μMのステアリン酸ナトリ
ウムを含む遊離コレステロール分析用試薬液を
200μ加え、最終液量を１mlにする。反応によ
つて生じた赤色キノンに基づく吸光度を測定して
遊離コレステロール濃度を求める。ステアリン酸
ナトリウムを含まない従来の試薬を用いたときに
は、前段の分析項目が中性脂肪の場合と同様に、
正誤差が非常に大である。それに対しこの実施例
によれば合成樹脂製反応容器を使用したことにと
もなう測定誤差はもたらされない。
以上説明したように本発明によれば、合成樹脂
製容器を種々の分析項目の反応に用いても、遊離
コレステロールを測定するときにリポプロテイン
リパーゼの残存による影響をなくすことができる
ので、高い測定精度の分析結果が得られる。[Table] As shown in Table 2, when conventional reagents that do not contain sodium oleate are used, the standard concentration is is 45mg/d
However, the measured value is over 140mg/d. Example 2 An analytical device having a row of transparent reaction vessels made of polystyrene resin was used. Similar to the analyzer of Example 1, multiple types of analytical items are reacted on one reaction vessel row, and the absorbance of the reaction liquid is measured by direct photometry. Into a specific container of the analyzer, a sample solution for total cholesterol is taken from the sampler and a total cholesterol reagent solution containing lipoprotein lipase, phosphate buffer, phenol, 4-aminoantipyrine, cholesterol oxidase and peroxidase is added, The reaction is allowed to occur, and the absorbance of the reaction solution is measured after a predetermined period of time. Thereafter, this reaction vessel is washed with water and subjected to a reaction for analyzing free cholesterol. Collect 20 μM of control serum into a specific reaction container that has been washed with water, and then add a reagent solution for free cholesterol analysis containing 20 μM sodium stearate instead of sodium olefinate in Table 1.
Add 200 μl to make a final volume of 1 ml. The free cholesterol concentration is determined by measuring the absorbance based on the red quinone produced by the reaction. When using a conventional reagent that does not contain sodium stearate, as in the case where the analysis item in the first step is neutral fat,
The correct error is very large. On the other hand, according to this embodiment, measurement errors due to the use of a synthetic resin reaction vessel are not introduced. As explained above, according to the present invention, even if synthetic resin containers are used for reactions for various analysis items, it is possible to eliminate the influence of residual lipoprotein lipase when measuring free cholesterol. Accuracy analysis results are obtained.
Claims (1)
て、遊離コレステロール以外の被検項目の分析に
使用した合成樹脂製反応容器を洗浄して使用する
こと、前記洗浄によつてもリポプロテインリパー
ゼが除去されずに該反応容器内に残存している場
合、オレイン酸ナトリウム、ステアリン酸ナトリ
ウム、ラウリル酸ナトリウム、およびパルミチン
酸ナトリウムからなる群から選ばれた化合物を含
む遊離コレステロール用分析試薬を試料と混合
し、該試薬と該試料とからなる反応液内での前記
化合物の濃度を２〜200μMとなすこと、および
前記反応液を光学的に測定して遊離コレステロー
ル濃度を求めることを特徴とする遊離コレステロ
ール分析法。1. In the method of analyzing free cholesterol, the reaction vessel made of synthetic resin used for the analysis of test items other than free cholesterol must be washed before use, and even after the washing, lipoprotein lipase is not removed and the reaction proceeds. If remaining in the container, an analytical reagent for free cholesterol containing a compound selected from the group consisting of sodium oleate, sodium stearate, sodium laurate, and sodium palmitate is mixed with the sample and the reagent and the A method for analyzing free cholesterol, characterized in that the concentration of the compound in a reaction solution comprising a sample is 2 to 200 μM, and the concentration of free cholesterol is determined by optically measuring the reaction solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP13847681A JPS5841357A (en) | 1981-09-04 | 1981-09-04 | Analysis of isolated cholesterol |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP13847681A JPS5841357A (en) | 1981-09-04 | 1981-09-04 | Analysis of isolated cholesterol |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5841357A JPS5841357A (en) | 1983-03-10 |
| JPS6350665B2 true JPS6350665B2 (en) | 1988-10-11 |
Family
ID=15222953
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP13847681A Granted JPS5841357A (en) | 1981-09-04 | 1981-09-04 | Analysis of isolated cholesterol |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5841357A (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0372093A (en) * | 1989-08-09 | 1991-03-27 | Mitsubishi Motors Corp | Acidic plating solution containing zinc |
| US9663816B2 (en) | 2012-04-27 | 2017-05-30 | Kyowa Medex Co., Ltd. | Method for measuring a component of a biological fluid and reducing the effect of interfering substances |
-
1981
- 1981-09-04 JP JP13847681A patent/JPS5841357A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5841357A (en) | 1983-03-10 |
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