EMA系列5——欧盟药品质量管理局-HPLC仪器确认指南(中英文)

QUALIFICATION OF EQUIPMENT
仪器确认
ANNEX 1: QUALIFICATION OF HPLC EQUIPMENT
附件1：HPLC仪器的确认
Introduction 概述
The present documentis the first Annex of the core document ―Qualification of Equipment‖, and
itshould be used in combination with it when planning, performing and documentingthe HPLC equipment qualification process.
本文件是核心文件―仪器的确认‖第1个附件，在计划、实施和记录HPLC仪器的确认过程时，
应将本文件与核心文件一起使用。

The core documentcontains the general introduction and the Level I and II of qualification,common to all type of instruments, and the present annex contains HPLCinstrument-related recommendations on parameters to be checked and thecorresponding typical acceptance Limits, as well as practical examples on the Methodologythat can be used to carry out these checks.
核心文件包括了第一级和第二级确认的通用介绍，适用于所有类型的仪器，本附件包括了HPLC仪器相关的需要检查的参数和相应典型的可接受标准的推荐，以及可用于进行这此检查的实用方法学举例。

When qualifyingHPLC equipment, it should be noted that it is acceptable to check at Level IIIand IV several of the mentioned parameters at the same time in a combined testprocedure (e.g. ―overall‖system performance test giving information on peakarea precision, retention time
precision, gradient reproducibility, etc).
在进行HPLC仪器确认时，应注意也可以对第三级和第四级几个提到的参数采用联合检测程序同时进行检查（例如―全面‖系统性能检查，同时给出峰面积精密度、保留时间精密度、梯度重复性等的信息）
Level III. Periodic and motivated instrument checks
第三级定期主动仪器检查
Examples of requirements for HPLC instruments anddetectors
HPLC仪器和检测器要求举例
HPLC SOLVENTDELIVERY SYSTEM 高压液相溶剂传输系统
The following testsare proposed for the periodic and motivated check of the HPLC solvent deliverysystem: flow rate and gradient test.
以下测试作为HPLC溶剂传输系统周期性主动检查：流速和梯度检查
FLOW RATE流速
Materials: 材料
Volumetric flask of5 or 10 ml
5ml或10ml的容量瓶
Calibratedchronometer
校正过的气压计
Settings: 设置
Mobile phase:degassed water
流动相：脱过气的水
No column (openend)*
无柱（开放式）*
Flow rate: adjustedbetween 0.5 and 3.0 ml/min
流速：调节在0.5到3.0ml/min
If high-pressuremixing systems are installed, this test has to be done on each solvent channel.
如果有安装高压混合系统，本测试应在每一溶剂通道进行。

* For certainequipment, e.g. in the case of low flow rates, the check would be performed byusing a column or a backpressure regulator.
* 对于一定的仪器，例如，流速较低时，采用一根柱子或背压调整器进行检查
Method方法:
Set the flow rate atan appropriate level and measure the time needed to fill the volumetric flaskup to the mark. Record the time needed.
将流速设定在一个合适的水平，测量将容量瓶装满至刻度的时间。

记录所用的时间。

f＝V /t f＝(V×60)/t
f ................. measured flow rate[ml/min]测量的流速
t ................. elapsed time to fill upto mark [s]充满至刻度所花的时间
V ............... volume of the volumetricflask [ml]容量瓶的体积
D＝100×(f－F)/F
D ...............deviation [%]偏差
F ............... adjusted flow rate [ml/min]调节后的流速
f ................. measured flow rate[ml/min]测量的流速
Limit限度s:±5%
GRADIENT COMPOSITIONAND RIPPLE 梯度组成和波动
Settings:设置
Stainless steelcapillary e.g. 2000 x 0.12 mminstalled instead of a column
不锈钢毛细管，例如2000X0.12mm，代替色谱柱安装于仪器
Detection:UV-Detector adjusted to 265 nm
检测器：紫外检测器调节至265nm
Mobile phase A:degassed water
流动相A：脱气水
Mobile phase B:degassed water containing 0.5% acetone
流动相B：含0.5%的脱气水
Flow rate: 1.0ml/min
流速：1.0ml/分钟
Method方法:
The test is carriedout in the following way by using a gradient program depending on the number ofsolvent channels and the configuration of the system:
按下列方法进行测试，根据通道数量不同使用下列不同系统参数和梯度进行洗脱
A-B
A-B and A-C
A-C, A-B and B-D
Start the test bypumping water for at least 10 min to equilibrate the system.
开始测试前，先用水平衡系统至少10分钟。

The zero % value atthe start of the test is the baseline. All steps are measured at the beginningof the horizontal part of the line either by software or manually on the paperprint using a liner. The height of the 100% water/acetone mixture is used asthe 100% value in the following calculation.开始时的0%值是基线值。

所有步骤均在开始时测量基线的水平部分，由软件或手动在纸上用衬垫打印。

100%水/丙酮混合物的高度在以下计算中作为100%值。

%H＝100* h/H
%H ............ calculated composition计算组成
h ................ height of the measuredline测量线的高度
H ............... height of the 100%water/acetone mixture line (mobile phase B)
100%水/丙酮混合物线的高度（流动相B）
d＝%H－G
d ................ deviation偏差
G ............... gradient compositionadjusted [% acetone/water solution mixture = mobile phase B]梯度组成调节【%丙酮/水溶液混合物= 流动相B】
Limit限度s:
Absolute deviation:±2 of the adjusted value
绝对偏差：调整值的±2
The ripple of thegradient composition is the percentage of noise of the 50% line from thegradient program.
梯度组分的波动为梯度程序噪声基线的50%
%R＝100* N /h50
%R ............ ripple波纹
h50 ............. height of the 50% line50%的线高度
N ............... height of the noise line,measured during 1 minute in the linear region噪声线高度，在线性区域1分钟测量的结果
Limit限度s: ≤0.2 %
HPLC INJECTOR 进样器
Volume precision andcarry-over are the tests proposed for the periodic and motivated check of theHPLC injector.
建议在HPLC进样器周期性主动检查中进行体积精密度和残留检查
VOLUME PRECISION ANDCARRY-OVER
体积精密度和残留
Solutions:溶液
Solvent A: methanol: water R, 60 : 40.
溶液A：乙醇：水（试剂）=60：40
Reference solution(a): dissolve 15.0 mg methyl-, ethyl-, and propylparabene in 100.0 ml ofsolvent A.
对照液（a）：取15.0mg乙基、甲基和尼泊金丙酯溶于100.0ml溶剂A中
Reference solution(b): Dilute 1.0 ml of reference solution (a) to 10.0 ml of solvent A.
对照液（b）：取1.0ml对照液（a）用溶于10.0ml溶剂A中
Reference solution(c): Dilute 1.0 ml of reference solution (b) to 100.0 ml of solvent A.
对照液（c）：取1.0ml对照液（b）用溶于100.0ml溶剂A中
Settings: 设置
Column: Lichrospher100 RP8, 5um, 125 x 4 mm, withoutprecolumn
色谱柱：Lichrospher 100 RP8, 5 um, 125 x 4 mm, 无衬管
Mobile phase:methanol : water = 60 : 40
流动相：乙醇：水= 60：40
Flow rate: 1.0ml/min
流动速度：1.0ml/分钟
Detection: 254 nm
检测器：254nm
Injection volume: 20μl
进样体积：20ul
Method方法:
Injection scheme: 进样序列
—6xreference solution (b)对照液（b）6针
—1x reference solution (a)对照液（a）1针
—1xsolvent A (blank injection 1)溶剂A（空白）1针
—1xreference solution (b)对照液（b）1针
—1xsolvent A (blank injection 2)溶剂A（空白2）1针
—1xreference solution (c)对照液（c）1针
Limit限度s:
Repeatability ofpeak areas: The relative standard deviation of the peak areas of all peaks inthe chromatogram obtained with the reference solution (b) should be≤1.0 %.
峰面积重复性：对照液（b）色谱图中所有峰的峰面积相对标准偏差应≤1.0 %
Carry-over: Thepercentage of the peak area corresponding to propylparabene in the blankinjection 1 does not exceed 0.5% of 10 times the peak area of thepropylparabene peak in the chromatogram obtained with the reference solution(b) injected after the blank injection.
残留：尼泊金丙酯在空白图谱中的峰面积不超过在空白后进样的对照液（b）中尼泊金丙酯的对应峰面积的10倍的0.5%
The percentage ofthe peak area corresponding to propylparabene in the reference solution (c) is0.9 – 1.1% of the peak area of the propylparabene peak in the chromatogramobtained with the reference solution (b) injected after the blank injection.
在对照液（c）中尼泊金丙酯对应的峰面积为空白后进样的对照液（b）中尼泊金丙酯峰面积的0.9-1.1%。

HPLC AUTOSAMPLER 液相色谱仪自动进样器
Thermostattingaccuracy and precision can be tested in the frame of the periodic and motivatedcheck of the HPLC Autosampler.
HPLC自动进样器的周期主动检查可以进行温度准确度和精密度检查THERMOSTATTINGACCURACY温度准确性
Materials: 材料
Calibrated temperatureprobe.
校正过的温度探头
Method方法:
Select a temperaturealong the operational or required temperature range of the equipment. Waituntil the system is equilibrated.
在可操作范围或要求的温度范围内选择并设定一个温度，等待系统平衡
By means of thecalibrated probe, measure the actual temperature in the autosampler and compareit to the selected temperature.
采用校正过的温度探头，测量自动进样器的实际温度，与设定的温度进行比较
Repeat the sameprocedure at different pre-selected points covering the temperature range.
在选定的温度范围内不同温度点重复相同的步骤
Limit限度s:
The actualtemperature may not differ more than±3°C with respect to the selected temperature.实际温度与设定的温度相比差值不超过±3°C
THERMOSTATTINGPRECISION温度准确性
Materials: 材料
Calibratedtemperature probe.
校正过的温度探头。

Method方法:
Select a temperaturealong the operational or required temperature range of the equipment. Waituntil the system is equilibrated.
选择一个操作温度或要求的仪器温度范围，等待系统平衡。

By means of thecalibrated probe, make ―n‖measurements over a pre-established period of pare the mean of the ―n‖ measurements to the selected temperature.
采用校正过的温度探头，在预设的时间内进行n次测量，比较n次测量值的平均值与设定的温度。

Limits限度:
The actualtemperature may not differ more than±3°C with respect to the selected temperature.
实际温度与设定的温度之间差值不超过±3°C
HPLC OVEN/COOLINGDEVICE 色谱仪柱温箱/降温设施
Thermostattingaccuracy is the parameter tested in this example of periodic and motivatedcheck of the HPLC oven/cooling device.
热力学准确性是在本例对HPLC柱温箱/降温设施周期主动检查的参数THERMOSTATTINGACCURACY温度准确性
Materials: 材料
Calibratedthermometer.
校正过的温度计
Method方法:
Set the column oventemperature to 40 °C,wait about 30 minutes to equilibrate the system, put a calibrated thermometerinto the oven and read the temperature after 10 minutes.
设定柱温箱温度为40 °C，等待约30分钟平衡系统，将校正过的温度计放入柱温箱中，在10分钟后读取温度。

Limits限度: 38 - 42°C.
HPLC UV/DADDETECTOR紫外/DAD检测器
The periodic andmotivated check of the HPLC UV/DAD detector can be performed by testing thelinearity and the wavelength accuracy.
对于HPLC紫外/DAD检测器的周期主动检查，可以对线性和波长准确性进行检查LINEARITY 线性
Solutions: 溶剂配制
Std. 1: 0.5 μg caffeine/1 ml methanol HPLC Grade
标准2：0.5 μg咖啡因/1ml色谱级甲醇
Std. 2: 1.0 μg caffeine/1 ml methanol HPLC Grade
标准2：1.0 μg咖啡因/1ml色谱级甲醇
Std. 3: 5.0 μg caffeine/1 ml methanol HPLC Grade
标准3：5.0 μg咖啡因/1ml色谱级甲醇
Std. 4: 25.0 μg caffeine/1 ml methanol HPLC Grade
标准4：25.0 μg咖啡因/1ml色谱级甲醇
Std. 5: 50.0 μg caffeine/1 ml methanol HPLC Grade
标准5：50.0 μg咖啡因/1ml色谱级甲醇
Std. 6: methanolHPLC Grade (blank)
标准6：色谱级甲醇（空白）
Std. 5: weigh 9.0 to11.0 mg caffeine and fill up to 200.0 ml with methanol HPLC 标准5：称取9.0至11.0mg咖啡因，用HPLC级别甲醇稀释至200.0ml Std. 4: dilute 50.0ml of Std. 5 to 100.0 ml with methanol
标准4：取50.0ml标准5溶液用甲醇稀释至100.0ml
Std. 3: dilute 10.0ml of Std. 5 to 100.0 ml with methanol
标准3：取10.0ml标准溶液5用甲醇稀释至100.0ml
Std. 2: dilute 20.0ml of Std. 3 to 100.0 ml with methanol
标准2：取20.0ml标准3溶液用甲醇稀释至100.0ml
Std. 1: dilute 10.0ml of Std. 3 to 100.0 ml with methanol
标准1：取10.0ml标准3用甲醇稀释至100.0ml
Settings: 设置
Column: RP-18 5 um30-50 x 2,1-4.6 mm or capillary 2000 mm x 0.12 mm ID
色谱柱：RP-185 um 30-50 x 2,1-4.6 mm or capillary 2000 mm x 0.12 mm ID Mobile phase:methanol HPLC Grade
流动相：色谱级甲醇
Oven temperature: 40 °C
柱温箱温度：40 °C
Flow rate: 1.0ml/min (adjusted by using 100% methanol)
流速：1.0ml/min（用100%甲醇调整）
Detection: 273 nm
检测波长：273nm
Injection volume:20μl
进样量：20ul
Method方法:
Injection scheme:
进样序列
2x blank
1x Std. 1
1x Std. 2
1x Std. 3
1x Std. 4
1x Std. 5
2针空白，标准1-5依序各一针
Limits限度: r2≥ 0.999
Remark: As this testemploys different test solutions to be injected, it covers also the check ofcorrect positioning vials in the autosampler.
注：由于此检测中要注入不同溶液，因此也包括了自动进样器样品瓶位置正确性的确认
WAVELENGTH ACCURACY 波长准确度
If there arebuilt-in test procedures for the determination and adjustment of wavelengthaccuracy, follow the instructions of the instrument manual.
如果内置检测程序可以检测和调节波长准确度，则根据操作手册进行操作
In all other casesuse the procedure described below.
在所有其它情况下采用以下程序
Solutions: 溶液
DAD: caffeine Std. 5from the linearity testing线性测试中咖啡因标准溶液5号
UV/VIS: 1.0 μg/ml anthracene solution in water蒽水溶液
Settings: 设置
Mobile phase: 15%acetonitrile in water
流动相：15%乙腈水溶液
Column: RP18, 5 m30-50 x 2,1-4.6 mm orcapillary 2.0 m x 0.12 mm ID
色谱柱：RP18，5μm,30-50×2.1-4.6mm或毛细管柱20m×0.12mm内径
Oven temperature: 40 °C
柱箱温度：40℃
Flow rate: 1.0ml/min (adjusted by using 15% acetonitrile in water)
流速：1.0ml/min（用15%的乙腈水溶液调节）
Detection: scan from230 nm to 290 nm (DAD)
检测：自230至290nm扫描（DAD）
Injection volume: 20μl
进样体积：20μl
Method方法:
DAD: Inject 20μl ofthe caffeine solution and record the spectrum. The maximum is at 272 nm and theminimum at 244 nm.
DAD: 咖啡因溶液进样20μl，记录色谱图，最大吸收波长为272nm，最小为244nm.
UV/VIS: fill thecell with the anthracene solution and change the wavelength from 248 to 254 nmin 1 nm steps; record the maximum of absorption. The theoretical value is 251nm.
UV/可见光：向池中注入蒽溶液，将波长以1nm的步距从248变化至254nm，记录最大吸收值。

理论值为251nm。

Limit限度s:≤2 nm
HPLC FLUORESCENCEDETECTOR 荧光检测器
The followingthree parameters are proposed for the performance of the periodic and motivatedcheck of the HPLC fluorescent detector:
下面三个参数建议作为荧光检测器性能周期自主检查
WAVELENGTH ACCURACY EXCITATION波长激发准确度
Method方法: 方法
Rinse and fill themeasuring cell with de-ionized water
用去离子水淋洗并充满检测池
Adjust theexcitation wavelength to 350 nm.
调节激发波长为350nm
Measure the emissionand subtract 397 nm (theoretical value).
测量发射波长减去397nm（理论值）
Limit限度限度s:±3 nm
WAVELENGTH ACCURACYEMISSION 波长发射准确度
Method方法: 方法
Rinse and fill themeasuring cell with de-ionized water
用去离子水淋洗并充满检测池
Adjust the emissionwavelength to 397 nm.
调节发射波长为397nm
Measure theexcitation and subtract 350 nm (theoretical value).
测量激发波长减去350nm（理论值）
Limit限度限度s:±3nm
SENSITIVITY 灵敏度
Solutions: 溶液
Quinine HCl.2H2O solution conc. 0.015μg/ml (=15 ppb)
二水盐酸喹啉溶液，浓度0.015μg/ml (=15 ppb)
The quinine solutionis prepared with the following mobile phase: dissolve 6.8 g of potassium dihydrogen phosphate R and 3.0 g of hexylamine R in 700 ml of waterR, adjust to pH 2.8 with dilute phosphoric acid R, add 90 ml of acetonitrile Rand dilute to 1000.0 ml with water R.
喹啉溶液用以下流动相制备：溶解6.8g磷酸二氢钾和3.0g己胺于700ml试剂级水中，用稀磷酸调pH至2.8，加90ml乙腈并用试剂级水稀释至1000.0ml.
Settings: 设置
The chromatographicconditions are set according to Ph. Eur. ―Quinine HCl‖ (01/2005:0018), test―Other cinchona alkaloids‖, with modified flow rate and acetonitrileconcentration. 色谱条件设定依据欧洲药典―盐酸喹啉‖（01/2005:0018），检测―金鸡纳生物碱‖，改变流速和乙腈
浓度。

Mobile phase: asabove
流动相：同上
Column: RP18, 5 μm,250× 4.6 mm
色谱柱: RP18, 5 μm, 250 ×4.6 mm
Flow rate: 1.2ml/min
流速：1.2 ml/min
Excitationwavelength: 350 nm
激发波长：350nm
Emission wavelength:397 nm
发射波长：397nm
Flow-cell volume: 8 μl(for this example, a Waters 2475 Multi Fluorescentie detector was used. Flow-cellvolume may vary depending on the instrument manufacturer)
流动池体积：8μl（此例中，使用的是Waters2475多荧光检测器，流动池体积可能根据仪器
厂商不同而不同）
Method方法:
Inject 10μl of thequinine solution and measure the peak height.
进样喹啉溶液10μl，测量峰高度
Inject 10μl of theblank and measure the peak height of the noise.
进样空白溶液10μl，测量噪音峰高
Divide the peakheight of the quinine solution by 3 times the peak height of the noise.
将喹啉溶液峰高除以3倍噪音峰高
Divide theconcentration of the quinine solution by the previously obtained factor.
将喹啉溶液浓度除以之前的系数
Limit限度限度s:≤ 0.5 ppb
HPLCELECTROCHEMICAL DETECTOR 高效液相电化学检测器
Accuracy andstability of the signal are the proposed parameters to be tested during theperiodic and motivated check of the HPLC electrochemical detector.
电化学检测器的信号准确度和稳定性是定期自主检测中需要测定的参数
ACCURACY ANDSTABILITY OF THE SIGNAL信号准确度和稳定性
Settings: 设定
Cell potential of adummy cell: 800 mV
空白池电压：800mv
Rise time filter:0.1 s
增加时间过滤：0.1s
Range: 0.1 nA
范围：0.1nA
Temperature: 30 °C
温度：30℃
Method方法:
Accuracy: Measurethe electric current and subtract 2.67 nA (theoretical value)
准确度：检测电流并减去2.67nA（理论值）
Stability: Measurethe noise over a period of 5 minutes
稳定性：连续5分钟测量噪音
Limit限度s:
Accuracy (cellcurrent): 0.05 nA
准确度（池电流）：0.05nA
Stability of thesignal (Noise): max. 2 pA or 20 mV
信号（噪音）稳定性：最大2pA或20mV
HPLC RID DETECTOR高效液相RID检测器
Signal to Noiseratio and drift over time are the parameters proposed for the periodic andmotivated check of the HPLC RID (refractive index) detector.高效液相RID（折光率）检测器的信噪比和对时间的漂移是定期自主检测中需要测定的参数
SIGNAL TO NOISERATIO 信噪比
Solutions溶液:
Standard solution:D-fructose concentrate solution at 4.0 mg/ml (dilute 200.0 mg fructose + 20 mlwater + 25.0 ml acetonitrile up to 50.0 ml with water for HPLC)
标准溶液：D-左旋糖溶液4.0mg/ml（将200.0mg左旋糖+20ml水+25.0ml乙腈，用色谱纯水稀释至50.0ml）
Settings设置:
Column: spherisorbNH2 (or equivalent) 250×4.6 mm or other
色谱柱：spherisorb NH2 (或相当的) 250×4.6 mm或其他
Oven temperature: 38°C
柱温箱温度：38℃
Flow rate: 1.0ml/min
流速: 1.0 ml/min
Injection volume: 20μl
进样量: 20 μl
Mobile phase: 0.253g sodium hydrogen phosphate R in 220 ml+ 780 ml acetonitrile
流动相：0.253g磷酸氢二钠溶于220ml+780ml乙腈
Method方法:
After equilibration,inject three times a blank solution of mobile phase over a run time where thesystem is stable. Measure the baseline noise over an appropriate period.
平衡后，以流动相为空白进样三针，系统稳定运行适当时间，测定适当时长内噪音基线The baseline noiseis accepted if the mean height of the three replicates is < 1000 μV.
可接受基线噪音：三次平行进样平均值< 1000 μV
To calculate thesignal to noise ratio, inject three times a solution of fructose at 0.4 mg/mland calculate the mean of the three replicates.
计算信噪比，左旋糖溶液0.4mg/ml进样三针，计算平均值
Limit限度s:S/N ≥10
限度：信噪比≥10
DRIFT OVER A DEFINEDPERIOD OF TIME 指定时长漂移
Method方法: 方法
Calculate the slopeof the amplitude of random variations in the detector’s signal over 1 minute.计算任意一分钟信号随机波动幅度的斜率
Limit限度s: ±0.1mV/min
限度：±0.1 mV/min
Alternatively, therequirement may be expressed in△RI/min or in % of full scale of theselected range.
也可以表达为△RI/min或者以选择量程的全范围%。

HPLC CD DETECTOR 高效液相色谱CD检测器
The following testsare proposed to perform the periodic and motivated check of the HPLC
CD(circular dichroism) detector.
建议配备有CD（圆二图谱）检测器的HPLC进行以下周期和主动检查
LINEARITY AND SIGNALTO NOISE RATIO
线性和信噪比
Solutions: 溶液
Reference solution(a): dissolve 25.0 mg D(-) pantolactone in 50.0 ml water
对照溶液（a）：溶解25.0mgD（-）泛酰内酯于50.0ml水中
Reference solution(b): dilute 2.0 ml of reference solution (a) to 10.0 ml with water
对照溶液（b）：取对照溶液（a）2.0ml用水稀释至10.0ml
Reference solution(c): dilute 4.0 ml of reference solution (a) to 10.0 ml with water
对照溶液（c）：取对照溶液（a）4.0ml用水稀释至10.0ml.
Reference solution (d):dilute 6.0 ml of reference solution (a) to 10.0 ml with water
对照溶液（d）：取对照溶液（a）6.0ml用水稀释至10.0ml.
Reference solution (e):dilute 8.0 ml of reference solution (a) to 10.0 ml with water
对照溶液（e）：取对照溶液（a）8.0ml用水稀释至10.0ml.
Reference solution (f):dilute 0.5 ml of reference solution (b) to 25.0 ml with water
对照溶液（f）：取对照溶液（b）0.5ml用水稀释至25.0ml.
Settings: 设定
Column色谱柱: C18, 150 ×4 mm,5 μm
Mobile phase流动相: acetonitrile乙腈: water水= 10 : 90
Flow流速: 1.0 ml/min
Detection检测波长: 225 nm
Injection volume进样量: 20 μl
Method方法:
Check the linearityof de CD- and UV-signal of D(-)pantolactone reference solution a,b,c,d,e.
检查CD和UV对D（-）泛酰内酯对照溶液a,b,c,d,e信号的线性关系
Measure the noise ofthe CD-signal of reference solution (f) between 0 – 10 min.
检查CD对对照溶液（f）在1-10分钟的噪声
- Calculate the absolute concentration (μg) in thecell
计算池中绝对浓度(μg)
- Calculate the signal-to-noise ratio (S/N) for 0.01 μgin the cell
计算池中0.01 μg信噪比（S/N）
-Calculatethe sensitivity with the calculated S/N and the specified S/N= 2 (0.01 x 2/
S/Ncalculated)
计算所得信噪比及信噪比为2(0.01 x 2/ S/N calculated)时的灵敏度
Limit限度s:
Linearity: Thelinearity of the calibration line obtained with reference solution (a,b,c,d,e)should be r > 0.999.
线性：由对照液（a,b,c,d,e）所得的校正曲线线性系统应>0.999。

Sensitivity: Thesensitivity at S/N= 2 should be better than 0.01μg.
灵敏度：信噪比为2时灵敏度应好于0.01μg。

S/N ratio: The Limitfor S/N is > 1.0 and the sensitivity should be 0.020 μg at maximum.
信噪比：信噪比为1.0时的限度和灵敏度最大为0.020μg。

DRIFT OVER A DEFINEDPERIOD OF TIME 指定时间内飘移值
Settings设定:
Column色谱柱: C18, 150 x 4 mm, 5μm
Mobile phase流动相: acetonitrile乙腈: water水= 10 : 90
Flow流速: 1.0 ml/min
Detection检测波长: 290 nm
Injection volume进样量: 20μl
Method方法:
Inject water andstop the flow after 5 minutes. Measure the CD-signal for 1 hour.
进水样，停泵，5分钟后测量CD信号，持续1小时。

Measure with thecursor the drift of the baseline between 5 and 65 min.
用标尺测量5到65分钟间的基线飘移
Limits限度:
Not more than 0.1mdeg/h
不超过0.1 mdeg/h
SPECTRA COMPARISON 谱图比较
Solutions溶液:
Reference solution(a): dissolve 5.0 mg dexamethasone in 10.0 ml 40% acetonitrile 对照溶液（a）：溶解5.0mg 地塞米松于10.0ml 40%乙腈中。

Settings设定:
Column色谱柱: C18, 150 x 4 mm, 5μm
Mobile phase流动相: acetonitrile乙腈: water水= 40 : 60
Flow流速: 1.0 ml/min
Detection检测波长: 230 nm
Injection volume进样量: 20ul
Method方法:
Compare themaxima/minima obtained at the Installation of the detector (see table).比较检测器安装结果（见下表）
Limits限度:
The maxima andminima may not differ more than± 4 nm.
最大和最小之间差异不可超过4nm。