Icp standard 4

The zinc and copper concentrations of the pre-treated fluid samples were measured by inductively coupled plasma optical emission spectrometry (ICP-OES 5100, Agilent Technologies, USA). The measurements were conducted in SVDV (Synchronous Vertical Dual View) mode, gaining intensity data from the axial and radial view, simultaneously. We applied an automatic sample introduction (SPS 4, Agilent Technologies, USA), and measured the samples in a randomized design. We performed measurements to generate a five-point calibration curve for the quantitative analysis of copper and zinc. Calibration solutions were diluted from a multi-element standard of 1000 mg/L (ICP standard IV, Merck, Germany) with 0.1 M nitric acid in ultrapure water. We expressed the trace element concentration of vaginal fluid samples in mg/L. The operating parameters of ICP-OES are described in Table 1.Table 1

The operating parameters of ICP-OES 5100 (Agilent Technologies)

ICP-OES operating parameters
Repetition	3
Pump speed	15 rpm
Uptake time	20 s, fast pump
Rinse time	30 s, fast pump
Read time	10 s
RF energy	1.20 kW
Stabilization time	7 s
View mode	SVDV
view height	0 mm
Nebulizer gas	0.70 l/min
Plasma gas	12.0 l/min

The elemental analysis of the pre-treated tissue and blood samples was carried out by inductively coupled plasma optical emission spectrometry (ICP-OES 5100, Agilent Technologies, Santa Clara, CA, USA). The measurements were conducted in SVDV (Synchronous Vertical Dual View) mode, gaining intensity data from the axial and radial view, simultaneously. An automatic sample introduction was applied (SPS 4, Agilent Technologies, Santa Clara, CA, USA), and the samples were measured in a randomized design [20 (link)]. We have performed measurements to generate a five-point calibration curve for the quantitative analysis of copper and zinc. We have diluted the calibration solutions from a multi-element standard of 1000 mg/l (ICP standard IV, Merck, Germany) with 0.1 M nitric acid in ultrapure water. We have expressed trace element concentration of vaginal tissues in milligrams per kilogram (mg/kg), and trace element concentration of whole blood samples in milligrams per liter (mg/l) [20 (link)].