仪器确认附件1HPLC确认中英文doc

PA/PH/OMCL (11) 04 - OMCL Guideline on qualification of HPLC equipment (Annex 1) 计算任意一分钟信号随机波动幅度的斜率

Limit限度s: ±0.1 mV/min 限度：±0.1 mV/min

Alternatively, the requirement may be expressed in △RI/min or in % of full scale of the selected 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 tests are 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 SIGNAL TO 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25.0mg D（-）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 linearity of 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 of the 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 the cell

计算池中绝对浓度(μg)

1st July 2011 Page 17 of 19

PA/PH/OMCL (11) 04 - OMCL Guideline on qualification of HPLC equipment (Annex 1) - Calculate the signal-to-noise ratio (S/N) for 0.01 μg in the cell 计算池中0.01 μg信噪比（S/N）

- Calculate the sensitivity with the calculated S/N and the specified S/N= 2 (0.01 x 2/

S/N calculated)

- 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: The linearity 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: The sensitivity 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 Limit for 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 DEFINED PERIOD 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 and stop 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 the cursor 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 0.1 mdeg/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

1st July 2011 Page 18 of 19

PA/PH/OMCL (11) 04 - OMCL Guideline on qualification of HPLC equipment (Annex 1) Detection检测波长: 230 nm Injection volume进样量: 20ul

Method方法:

Compare the maxima/minima obtained at the Installation of the detector (see table). 比较检测器安装结果（见下表） CD max 最大 CD min 最小 UV max 最大 222 nm 224 nm 236 nm 230 nm 252 nm 284 nm Limits限度:

The maxima and minima may not differ more than± 4 nm. 最大和最小之间差异不可超过4nm。

1st July 2011 Page 19 of 19