【正文】 at least 3 mm (HPTLC) to 5 mm (TLC) above the level of the developing solvent—and from the sides of the plate. Apply the solutions on a line parallel to the lower edge of the plate with an interval of at least 10 mm (5 mm on HPTLC plates) between the centers of spots or 4 mm (2 mm on HPTLC plates) between the edges of bands, and allow to dry. 步驟：使用規(guī)定體積的供試溶液和標(biāo)準(zhǔn)溶液，分成足夠多的小份進(jìn)行點(diǎn)樣，以得到直徑 2 至 5mm（在高效薄層色譜板上為 1 至 2mm）的圓形斑點(diǎn)或者 10至 20mm 乘 1 至 2mm（在高效薄層色譜板上為 5 至 10mm 乘 至 1mm）帶狀斑，并與下邊緣（在色譜過程中，點(diǎn)樣位置必須在展開溶劑水平面至少3mm（高效薄層色譜法）至 5mm（薄層色譜法）以上）和薄板的側(cè)面保持適當(dāng)距離。將這些溶液點(diǎn)在平行于該薄板下邊緣的直線上，并且斑點(diǎn)的中心之間距離至少 10mm（在高效薄層色譜板上 5mm）或者帶狀斑邊緣之間的距離至少4mm（在高效薄層色譜板上 2mm），并待其變干。Ascending Development— Line at least one wall of the chromatographic chamber with filter paper. Pour into the chromatographic chamber a quantity of the mobile phase sufficient for the size of the chamber to give, after impregnation of the filter paper, a level of depth appropriate to the dimension of the plate used. For saturation of the chromatographic chamber, close the 17 / 75lid, and allow the system to equilibrate. Unless otherwise indicated, the chromatographic separation is performed in a saturated chamber. 上行展開 ：將色譜室內(nèi)至少一側(cè)內(nèi)壁襯以濾紙。將相對(duì)色譜室大小而言足夠量的流動(dòng)相倒入該室，以便在加入濾紙之后在能合適于薄板大小的深度。為了飽和色譜室，蓋上蓋子，并使該系統(tǒng)達(dá)到平衡。除非另外指出，色譜分離需在飽和的色譜室內(nèi)進(jìn)行。Place the plate in the chamber, ensuring that the plate is as vertical as possible and that the spots or bands are above the surface of the mobile phase, and close the chamber. The stationary phase faces the inside of the chamber. Remove the plate when the mobile phase has moved over the prescribed distance. Dry the plate, and visualize the chromatograms as prescribed. For twodimensional chromatography, dry the plates after the first development, and carry out a second development in a direction perpendicular to that of the first development.將薄板置于色譜室中，確保該薄板盡可能垂直，并且斑點(diǎn)或帶狀斑在流動(dòng)相的表面之上，然后關(guān)閉該室。固定相面向該室的內(nèi)部。當(dāng)流動(dòng)相已經(jīng)移動(dòng)超過了規(guī)定的距離，則取出該薄板。干燥該薄板，并按照規(guī)定使得色譜圖顯色。對(duì)于雙向色譜法，在第一次展開之后干燥薄板，然后進(jìn)行在垂直于第一次展開的方向上進(jìn)行第二次展開。Horizontal Development— Introduce a sufficient quantity of the developing solvent into the reservoir of the chamber using a syringe or pipet. Place the plate horizontally in the chamber, connect the mobile phase direction device according to the manufacturer39。s instructions, and close the chamber. If prescribed, develop the plate starting simultaneously at both ends. Remove the plate when the mobile phase has moved over the distance prescribed in the monograph. Dry the plate, and visualize the chromatograms as prescribed. 18 / 75水平展開 ：使用注射器或者移液器，將充足數(shù)量的展開溶劑加入至該室的貯液池中。將該薄板水平置于該室中，根據(jù)生產(chǎn)商的說明連接流動(dòng)相導(dǎo)向設(shè)備，并關(guān)閉該室。如有規(guī)定，則在兩端同時(shí)開始展開該薄板。當(dāng)流動(dòng)相已經(jīng)移動(dòng)超過了該各論中規(guī)定的距離，則取出該薄板。干燥該薄板，并按照規(guī)定將色譜圖顯色。For twodimensional chromatography, dry the plates after the first development, and carry out a second development in a direction perpendicular to that of the first development.對(duì)于雙向色譜法，在第一次展開后干燥薄板，并在垂直與第一次展開的方向上進(jìn)行第二次展開。Detection— Observe the dry plate first under shortwavelength UV light (254 nm) and then under longwavelength UV light (365 nm) or as stated in the monograph. If further directed, spray, immerse, or expose the plate to vapors of the specified reagent, heat the plate when required, observe, and pare the test chromatogram with the standard chromatogram. Document the plate after each observation. Measure and record the distance of each spot or zone from the point of origin, and indicate for each spot or zone the wavelength under which it was observed. Determine the RF values for the principal spots or zones (see Glossary of Symbols). 檢視 ：首先在短波長紫外光下（254nm ），然后在長波長紫外光（ 365nm）下，或者按照在該各論中的論述，觀察干燥的薄板。如果有進(jìn)一步的規(guī)定，噴灑、浸漬、或?qū)⒈“灞┞队谥付ㄔ噭┑恼羝?，?dāng)需要時(shí)加熱薄板，觀察并將供試色譜圖與標(biāo)準(zhǔn)色譜圖進(jìn)行比較。在每次觀察之后將該薄板記錄并存檔。測(cè)量并記錄從起始點(diǎn)到每一個(gè)斑點(diǎn)或區(qū)域的距離，并且指明在觀察每一個(gè)斑點(diǎn)或區(qū)域時(shí)所用的波長。確定基本斑點(diǎn)或區(qū)域的 RF 值（見 符號(hào)術(shù)語表 ）。Quantitative Measurement— Using appropriate instrumentation, substances separated by TLC and responding to ultravioletvisible (UVVis) irradiation 19 / 75prior to or after derivatization can be determined directly on the plate. While moving the plate or the measuring device, the plate is examined by measuring the reflectance of the incident light. Similarly, fluorescence may be measured using an appropriate optical system. Substances containing radionuclides can be quantified in three ways: (1) directly by moving the plate alongside a suitable counter or vice versa。 (2) by cutting the plates into strips and measuring the radioactivity on each individual strip using a suitable counter。 or (3) by scraping off the stationary phase, dissolving it in a suitable scintillation cocktail, and measuring the radioactivity using a liquid scintillation counter (see Radioactivity 821 ). 定量測(cè)量：使用適當(dāng)?shù)膬x器，被 TLC 分離并且在衍生作用之前或之后對(duì)紫外－可見光（UVVis）照射有響應(yīng)的物質(zhì)可以直接在薄板上進(jìn)行測(cè)定。當(dāng)移動(dòng)該薄板或測(cè)量設(shè)備時(shí)，通過測(cè)量入射光線的反射系數(shù)來檢查該薄板。同法，可以使用適當(dāng)?shù)墓鈱W(xué)系統(tǒng)來測(cè)量熒光。含有放射性核素的物質(zhì)可以通過以下 3 種方法進(jìn)行定量：(1)直接通過沿著適當(dāng)?shù)挠?jì)數(shù)器移動(dòng)該薄板，反之亦然； (2)通過將該薄板分隔成條狀并使用適當(dāng)?shù)挠?jì)數(shù)器來測(cè)量單個(gè)條狀物上的輻射性；或者，(3)通過刮下固定相，溶解于適當(dāng)?shù)拈W爍雞尾酒，并使用液體閃爍計(jì)數(shù)器來測(cè)量放射性（見 放射性 821）。The apparatus for direct quantitative measurement on the plate is a densitometer that is posed of a mechanical device to move the plate or the measuring device along the xaxis and the yaxis, a recorder, a suitable integrator or a puter。 and, for substances responding to UVVis irradiation, a photometer with a source of light, an optical device capable of generating monochromatic light, and a photo cell of adequate sensitivity, all of which are used for the measurement of reflectance. In the case where fluorescence is measured, a suitable filter is also required to prevent the light used for excitation from reaching the photo cell while permitting the emitted light or 20 / 75specific portions thereof to pass. The linearity range of the counting device must be verified.用于薄板上直接定量測(cè)量的儀器是一個(gè)光密度計(jì)，其由可以移動(dòng)薄板的設(shè)備或可以沿 X 軸和 Y 軸移動(dòng)薄板進(jìn)行測(cè)量的機(jī)械設(shè)備、記錄器、適當(dāng)?shù)姆e分儀或計(jì)算機(jī)，并且待測(cè)物質(zhì)對(duì)響應(yīng)紫外－可見光照射有響應(yīng)，帶有光源的光度計(jì)、能夠產(chǎn)生單色光的光學(xué)設(shè)備，以及有足夠敏感度的光電管組成，所有上述設(shè)備組合起來用于測(cè)量反